GJRMI - Volume 2, Issue 4, April 2013 by Dr. Hari Venkatesh.K.Rajaraman

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ISSN 2277 â&#x20AC;&#x201C; 4289 www.gjrmi.com Editor-in-chief Dr Hari Venkatesh K Rajaraman

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INDEX – GJRMI, Vol.2, Iss. 4, April 2013 MEDICINAL PLANTS RESEARCH Laboratory Sciences & Ethno-Botany SCREENING FOR ANTIMICROBIAL ACTIVITY OF SOME PLANTS FROM SAUDI FOLK MEDICINE Abdallah Emad M, El-Ghazali Gamal E

189–197

Botany DIVERSITY AND AVAILABILITY STATUS OF ETHNOMEDICINAL PLANTS IN THE LOHBA RANGE OF KEDARNATH FOREST DIVISION (KFD), GARHWAL HIMALAYA Ballabha Radha, Singh Dinesh, Tiwari J K, Tiwari P

198–212

Ophthalmology & Pharmacology ANTICATARACT ACTIVITY OF ERVATAMIA CORONARIA CHEMICALLY INDUCED CATARACTOGENESIS IN RATS Rathnakumar K, Jaikumar S, Duraisami R, Sengottuvelu S

LEAF

EXTRACT

213–218

Ethno-Botany STUDIES ON TRADITIONAL HERBAL PEDIATRICS PRACTICES IN JAISINGHPUR, DISTRICT KANGRA (HIMACHAL PRADESH, INDIA) Rawat Dhiraj S, Kharwal Anjna D

219–230

Bio-Chemistry LIVER ENZYMES AND ITS ASSOCIATION WITH AGE AND SEX IN SICKLE CELL ANAEMIA PATIENTS AND HAEMOGLOBIN S TRAIT CARRIES. Chuku L C, Chinaka N C

231–237

Life Sciences CHELIDONIUM MAJUS L. - A REVIEW ON PHARMACOLOGICAL ACTIVITIES AND CLINICAL EFFECTS Biswas Surjyo Jyoti

238–245

INDIGENOUS MEDICINE Ayurveda – Dravya Guna A DETAILED PHARMACOGNOSTICAL EVALUATION ON LEAF OF OLAX SCANDENS ROXB. Naik Raghavendra, Borkar Sneha D, Harisha C R, Acharya R N

246–253

Ayurveda – Rachana Sharira A CLINICAL EVALUATION ON RUJAKARA MARMA WITH SPECIAL REFERENCE TO PAIN THRESHOLD Benjwal Shobha

254–258

Ayurveda – Dravya Guna PHARMACOGNOSTICAL AND PRELIMINARY PHYTOCHEMICAL INVESTIGATIONS ON DIFFERENT PARTS OF BULBOPHYLLUM NEILGHERRENSE WIGHT. -AN ORCHID USED IN FOLK MEDICINE. Kumari Harshitha, Nishteswar K, Harisha C R

259–269

Ayurveda – Dravya Guna A COMPARATIVE ACUTE TOXICITY EVALUATION OF ASHOKA KSHEERAPAKA PREPARED FROM TWO DIFFERENT SPECIES OF SARACA (S. ASOCA & S. THAIPINGENSIS) Chavan S S, Gamit R V, Ashok B K, Shukla V J, Das P, Ravishankar B

COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – BRANCH OF KEBUKA [CHEILOCOSTUS SPECIOSUS (J. KONIG) C. SPECHT], OF THE FAMILY COSTACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT, KARNATAKA, INDIA

270–277

Research article SCREENING FOR ANTIMICROBIAL ACTIVITY OF SOME PLANTS FROM SAUDI FOLK MEDICINE Abdallah Emad M1*, El-Ghazali Gamal E2 1, 2

Department of Laboratory Sciences, College of Science and Arts, Al-Rass, P.O. Box 53, Qassim University, Saudi Arabia. *Corresponding author: emad100sdl@yahoo.com

Received: 24/02/2013; Revised: 28/03/2013; Accepted: 31/03/ 2013

ABSTRACT Methanolic extract of nine medicinal plants from Saudi folk medicine (Tamarix aphylla, Dactyloctenium aegyptium, Francoeuria crispa, Rhazya stricta, Trichodesma africanum, Haloxylon salicornicum, Echinops spinosissimus, Zygophyllum simplex and Blepharis ciliaris) were examined for their phytochemical compounds and antimicrobial potential against seven standard bacteria (Proteus vulgaris NCTC 8196, Escherichia coli ATCC 25922, Bacillus cereus NCTC 8236, Salmonella typhi NCTC 0650, Klebsiella pneumonia ATCC 53651, Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 25923) and one standard fungus (Candida albicans ATCC 7596). The phytochemical analysis showed presence of some active principles which correlates with the antimicrobial activity of some plant extracts. Most plants showed some degree of antimicrobial activity. However, the methanol extracts of Rhazya stricta, Francoeuria crispa and Blepharis ciliaris respectively, recorded the maximum antimicrobial activities compared to Chloramphenicol as antibacterial and Clotrimazole as antifungal antibiotic. The results of this investigation support the use of these plants in Saudi folk medicine for treatment of ailments caused by microorganisms. KEYWORDS: Medicinal plants, folk medicine, phytochemical, antimicrobial

Cite this article: Abdallah Emad M, El-Ghazali Gamal E (2013), SCREENING FOR ANTIMICROBIAL ACTIVITY OF SOME PLANTS FROM SAUDI FOLK MEDICINE, Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 189â&#x20AC;&#x201C;197

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 189â&#x20AC;&#x201C;197

INTRODUCTION No doubt that what we called it folk, traditional or alternative medicine was the main source for remedies since antiquity. On the other side, what we called modern medicine is now facing challenge due to its failure to convoy the development of diseases. Accordingly, there is a serious need to go back to the Mother Nature and its rich sources of natural remedies. It is well known that, man has been employing natural products as remedies since times immemorial, this knowledge accumulated and passed from generation to generation. Thus, the ethno-botanical knowledge about drugs considered as a basic source for new therapeutics.

Tamarix aphylla, Dactyloctenium aegyptium, Francoeuria crispa, Rhazya stricta, Trichodesma africanum, Haloxylon salicornicum, Echinops spinosissimus, Zygophyllum simplex and Blepharis ciliaris are chosen to study because they are already being utilized in folk medicine as antimicrobials (in treating wounds, sores, inflammations, cough and cold, etc.) by native inhabitants and Bedouin at Al-Rass province, Qassim district, Saudi Arabia (El-Ghazali et al., 2010.) In the present study, the antimicrobial activities beside the preliminary phytochemical investigation of the above mentioned plants were evaluated. MATERIALS AND METHODS

Recently, the misuse of synthetic antibiotics has promoted the emergence of antibioticresistant pathogens, including multidrug resistant strains (Kumar and Schweizer, 2005). This predicament, impose to search for other alternatives that are not centered upon standard antibiotics therapy, or we risk the possibility of eventually having no defense against these antibiotic-resistant pathogens (Treadway, 1998). Medicinal plants, particularly those employed in folk medicine could be a promising alternative, as it could gives a new source of antimicrobial agents with possibly novel mechanisms of action (Runyoro et al., 2006). The first step towards discovering novel antimicrobials is the screening of such plants.

Plant material

Saudi Arabia has a hot desert climate and rainfall is scarce in most parts of the country. The flora of Saudi Arabia as well as the other countries in the peninsula has been neglected for a long time due to its arid climate. The first attempt to cover the flora of Saudi Arabia was in 1974 (Alfarhan et al., 1998). However, folk medicine, including medicinal herbs, occupies a significant part of Saudi Arabiaâ&#x20AC;&#x2122;s heritage and it is widely practiced until now (Al-Essa et al., 1998). Although, the growing development in healthcare in Saudi Arabia which based on the western modern medicine decreases the public interest in the traditional medicine, particularly in towns.

All microbial strains, 7 reference bacterial strains representing the gram negatives (Escherichia coli ATCC 25922, Klebsiella pneumonia ATCC 53651, Pseudomonas aeruginosa ATCC 27853, Proteus vulgaris NCTC 8196, Salmonella typhi NCTC 0650) and gram positives (Bacillus cereus NCTC 8236, Staphylococcus aureus ATCC 25923), and one reference fungal strain (Candida albicans ATCC 7596) were obtained from the stock culture of the microbiology laboratory, Medicinal and Aromatic Plants Research Institute, Khartoum, Sudan.

Plants were collected based on information of previous ethnobotanical survey on medicinal plants used by native nomadic people (ElGhazali et al., 2010). In this study, nine selected plants claimed to be used as antimicrobials in folk medicine (Antidiarrheal, antiseptic, anti-inflammatory, anti-cold and cough, and in wound treatment) were collected from Al-Rass province, Qassim district, Saudi Arabia. All plants were identified by Gamal E. El-Ghazali (Taxonomist). Information regarding these plants is shown in Table 1. Microbial strains

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Inoculum preparation Active cultures were prepared by inoculating the stock culture into sterile bottles containing Nutrient broth (MAST Laboratories LTD, UK) for bacteria and Malt extract broth (OXOID, UK) for fungus and incubated for 24 h at 37°C and 72 h at 25°C respectively. The turbidity of actively growing bacterial suspension was adjusted to match the turbidity standard of 0.5 McFarland. This turbidity is equivalent to approximately 1–2 × 108 CFU/ml 5 for bacteria and about 2.0 × 10 spore/ml for fungi. This suspension was used for the antimicrobial examination. Plant extracts Plant parts (leaves or whole plant) were washed with distilled water and air dried in shade inside the laboratory for about 2–3 weeks until totally dried and grounded with a grinder machine. 50 g of each ground material was soaked in 500 ml of methanol, for at least 72 h with frequent shakings. The samples were filtered using Whattman No.1 filter paper (Whatman limited, UK). The filtrate was evaporated to dryness under reduced pressure at 40 oC. All the extracts (in powder form) were kept in refrigerator in dark bottles until used (Samie et al., 2005). Phytochemical screening Screening for some active phytochemical principles claimed having antimicrobial activity were undertaken as described by Edeoga et al., (2005), Krishnaiah et al., (2009) and Abdallah et al., (2009) for detection of tannins, saponins, flavonoids, terpenoids, phenolic compounds, alkaloids and anthraquinones. Antimicrobial assay The antimicrobial activity of the methanol extract of the nominated plant extracts were evaluated using agar-well diffusion method as mentioned by Abdallah et al., (2012), with minor modifications. The dry methanolic extracts of plants under study were reconstructed with 70% methanol to make a final

concentration 100 mg/ml and filtered using 0.22 μm pore-size black polycarbonate filters (Millipore). To a sterile Petri-dish (Size 100 mm), 25 ml of molten Nutrient Agar (MAST Laboratories Ltd, UK) or Potato Dextrose agar (Oxoid Ltd, UK) was poured and left to solidify. Fresh working cell suspensions (Bacteria or fungus) were prepared and adjusted to 0.5 McFarland's standard. Then from each microorganism, 100 μl was spread onto the surface of the plates of Nutrient Agar for bacteria or Potatoes Dextrose Agar for fungi. After about 15 min., 6 mm wells were punched into the agar using a sterile cork borer. Afterwards, 100 μl (10000 μg/wells) from each concentration was loaded into the wells of the previously prepared plates and incubated for 24 h at 37°C for bacterial strains and 72 h at 25°C for fungus. Chloramphenicol 5 mg/ml (50 μg/wells) (Riyadh Pharma. Co. Ltd, SA) and Clotrimazole 10 mg/ml (100 μg/wells) (Pharco Pharmaceuticals, Egypt) were employed as antibacterial and antifungal positive controls, respectively. 70% methanol was employed as a negative control. Each test was repeated twice and the mean zone of inhibition was recorded. RESULTS AND DISCUSSION Nowadays, the natural products and medicinal plants are a subject of great global interest for the discovery of new antimicrobial agents (Sashikala et al., 2009). This could be related to the recent failure of antibiotics against the dramatic emerging of the multidrug resistant pathogens in addition to the rapid spread of the new infections (Abdallah, 2011). The ethnobotanical information of the studied plants and their usage in the folk medicine at Al-Rass province, Saudi Arabia are summarized in Table 1. The ethnobotanical approach has advantages over the random screening, as this approach depends on the human experience with diseases through generations. The results of the phytochemical screening of the nominated plant extracts (Methanol extracts) showed that, all the methanolic

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extracts of the plants exhibited presence of at least two or more of these phytochemical compounds (Table 2). The phytochemical compounds studied in this investigation having antimicrobial potency, as reported by many researchers such as Watt and Pretorius (2001) for tannin and phenolic compounds, Deeni and Sadiq (2002) and Rohit et al. (2012) for

anthraquinones and saponins, RÎŻos and Recio (2005) for alkaloids, flavonoids and terpenoids. It is known that the phytochemical compounds, which are secondary metabolic products in plants, produce some biological activities in human and animal and responsible for their use as a drug (Sofowora, 1984).

Table 1: Ethnobotanical information of some selected medicinal plants from Al- Rass province Scientific name

Family

Common names Taramisk, Atheltrep, Salt cedar, Athel, Athel pine Crowfoot grass, Star grass, Beach wire grass, Button grass, Buck grass Francoeuria

Part tested Leaves

Traditional uses

Tamarix aphylla (L.) Karsten.

Tamaricaceae

Dactyloctenium aegyptium (L.) Willd.

Poaceae (Graminae)

Francoeuria crispa (Forssk.) Cass.

Asteraceae (Compositae)

Rhazya stricta Decne.

Apocynaceae

Trichodesma africanum (L.) Lehm. Haloxylon salicornicum (Moq.) Boiss. Echinops spinosissimus Turra. Zygophyllum simplex L. Blepharis ciliaris (L.)B.L. Burtt.

Boraginaceae

Harmal, Senhwar, Dogbane. NA*

Chenopodiaceae

NA*

Whole plant Its smoke breathed to treat cold

Asteraceae

NA*

Leaves

Zygophyllaceae

NA*

Whole plant In treating ophthalmia

Acanthaceae

NA*

Whole plant

Anti-inflammatory and in treating wounds

Whole plant In treating wounds

Whole plant Its smoke breathed to treat the upper thorax inflammations Leaves Dried leaves used in healing wounds Whole plant Anti-cold

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In treating the upper thorax infections.

In treating wounds and in renal disorder. *NA: Not available.

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Table 2: Qualitative analysis of the phytochemicals of the medicinal plants Plant Tamarix aphylla Dactyloctenium aegyptium Francoeuria crispa Rhazya stricta Trichodesma africanum Haloxylon salicornicum

Tan ‒ ‒

Sap + ±

+ ‒ + ‒

‒ ‒ + +

Phytochemical compounds* Fla Ter Phen + + ‒ + ‒ ‒ + ‒ ± ‒

+ + + +

Alk ± +

Anth ‒ ‒

+ + ‒ +

+ + ‒ ‒

+ + ± +

Echinops spinosissimus ± + + + ± ‒ ‒ Zygophyllum simplex + + + + + ± ± Blepharis ciliaris + + + + + + ‒ Negative control (D.W.) ‒ ‒ ‒ ‒ ‒ ‒ ‒ * Tan= Tannins, Sap= Saponins, Fla= Flavonoids, Ter= Terpenoids, Phen= Phenolic compounds, Alk= Alkaloids, Anth= Anthraquinones. + = Presence, − = Absence, ± = Weak reaction, D.W. = Distilled water.

Table 3: Screening of the methanol extracts of some plant species for antimicrobial activity of Gram-negative bacteria Mean zone of inhibition (mm) of microorganisms (Mean±SEM)**

Tested*

Tamarix aphylla Dactyloctenium aegyptium Francoeuria crispa Rhazya stricta Trichodesma africanum Haloxylon salicornicum Echinps spinosissimus Zygophyllum simplex Blepharis ciliaris Chloramphenicol 5 mg/ml

11.0± 2.0 12.5 ± 0.5 18.0 ± 1.0 20.5 ± 0.5 9.5 ± 0.5 11.5 ± 1.5 12.5 ± 1.5 14.5 ± 0.5 18.5 ± 0.5 26.0 ± 1.0

9.0 ± 1.0 8.0 ± 0.0 19.0 ± 1.0 19.5 ± 0.5 10.5 ± 0.5 13.0 ± 0.0 12.0 ± 0.0 10.0 ± 1.0 14.5 ± 0.5 13.7 ± 1.05

8.5 ± 0.5 12.0 ± 1.0 15.0 ± 0.0 20.0 ± 2.0 12.5 ± 0.5 12.0 ± 2.0 12.0 ± 1.5 8.5 ± 0.5 16.0 ± 0.0 22.0 ± 0.2

10.5 ± 1.5 11.5 ± 1.5 15.5 ± 0.5 24.0 ± 1.0 10.5 ± 0.5 12.0 ± 0.0 10.5 ± 0.5 9.5 ± 0.5 15.0 ± 2.0 24.0 ± 1.0

15.5 ± 0.5 13.0 ± 0.0 16.0 ± 0.0 9.5 ± 0.5 14.0 ± 0.0 15.5 ± 0.5 13.0 ± 0.0 13.0 ± 0.0 14.5 ± 1.5 16.5 ± 1.5

* Plants tested are as methanol extracts at 100mg/ml, Chloramphenicol as antibacterial at 5 mg/ml. **Mean ± Standard error of means (SEM ), mm=millimeter; Microorganisms: Pr = Proteus vulgaris NCTC 8196, Ec = Escherichia coli ATCC 25922, Kp = Klebsiella pneumonia ATCC 53651, Ps = Pseudomonas aeruginosa ATCC 27853, Sa = Staphylococcus aureus ATCC 25923.

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 189–197

Table 4: Screening of the methanol extracts of some plant species for antimicrobial activity of Gram-positive bacteria and fungus

Tested*

Mean zone of inhibition (mm) of microorganisms (Mean±SEM)** Bs

Sal

Cand

12.0 ± 2.0 11.0 ± 2.0 11.5 ±1.5 10.0 ± 1.0 13.5 ± 1.5 12.5 ± 1.5 13.0 ± 0.0 9.5 ± 0.5 11.5 ± 0.5 21.0 ± 1.0 ‒

11.0 ±0.0 9.5 ± 0.5 10.5 ± 0.5 19.0 ± 0.5 11.5 ± 1.5 12.5 ± 2.5 13.5 ± 0.5 13.0 ± 2.0 14.5 ± 0.5 13.0 ± 1.0 ‒

6.5 ± 0.5 11.5 ± 0.5 18.0 ± 1.0 14.0 ± 1.0 17.5 ± 0.5 14.5 ± 0.5 15.0 ± 0.0 13.5 ± 0.5 18.0 ± 1.0 ‒ 13.0 ± 1.0

* Plants tested are as methanol extracts at 100 mg/ml, Chloramphenicol as antibacterial at 5 mg/ml and Clotrimazole as antifungal at 10 mg/ml, ‒ = Not tested. **Mean ± Standard error of means (SEM), mm=millimeter; Microorganisms: Bs = Bacillus cereus NCTC 8236, Sal = Salmonella typhi NCTC 0650, Cand = Candida albicans ATCC 7596.

The antibacterial and antifungal activities of the methanolic plant’s extracts are shown in Tables 3 and 4. Methanol as a solvent is the most commonly used solvents for preliminary studies of antimicrobial activities in plants (Das et al., 2010). In general, most plants showed some degree of antimicrobial activity against tested microorganisms. However, Rhazya stricta, Francoeuria crispa and Blepharis ciliaris respectively, recorded the highest antimicrobial activity among other plants, even higher than that was recorded by the commercial antibiotic itself (Tables 3 and 4). E. coli was much susceptible to Rhazya stricta (19.5 ± 0.5), Francoeuria crispa (19.5 ± 0.5 mm) and Blepharis ciliaris (14.5 ± 0.5 mm) when compared to Chloramphenicol (13.7 ± 1.05 mm). Similarly, Salmonella typhi was highly susceptible to Rhazya stricta (19.0 ± 0.5 mm) and Blepharis ciliaris (14.5 ± 0.5 mm), compared to Chloramphenicol (13.0 ± 1.0 mm). Also, the susceptibility of Pseudomonas aeruginosa to

the methanol extract of Rhazya stricta was similar to that of Chloramphenicol (24.0 ± 1.0 mm). Our results regarding the antibacterial potential of the methanol extract of Rhazya stricta are in harmony with Ahmad et al., (2004) who mentioned that the crude ethanolic extract of Rhazya stricta exhibited a considerable antibacterial activity against a wide range of gram-positive and gram negative bacteria, the susceptibility of some of these bacteria to this plant extract were higher than that of the antibiotic Co-trimoxazole. Same result recorded by Staphylococcus aureus where the inhibition zones with the methanol extract of Francoeuria crispa almost equal to its inhibition zone with Chloramphenicol (Tables 3 and 4). Similarly, the results of this study regarding the methanol extract of Francoeuria crispa (Tables 3 and 4) are in agreement with the findings of El-Kamali and Mahjoub (2009) there were found that the most susceptible bacteria were E. coli and P. vulgaris and Staphylococcus aureus and the

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least susceptible were Salmonella para typhi and B. subtillis. Also, Blepharis ciliaris was reported in literature as a plant of antimicrobial properties (El-Shanawany et al., 2012) agrees with the findings of this investigation. The antibiotic Chloramphenicol became available commercially in 1948 and it was active against all gram-positive and many gram-negative bacteria at that time. However, within a few years bacterial resistance to Chloramphenicol was recorded (Shaw, 1984). Since Rhazya stricta, Francoeuria crispa and Blepharis ciliaris exhibited maximum antibacterial activities higher than the Chloramphenicol against some bacteria, it is believed that these plants when extracted and purified may lead to new effective antibacterial drugs. As shown in Tables 3 and 4, the results of the antifungal activities showed that, most plant extracts exhibited antifungal activity higher than the antifungal agent (Clotrimazole 10 mg/ml). The methanol extract of Francoeuria crispa and Blepharis ciliaris (18.0 ± 1.0 mm), besides Tricodesma africanum (17.5 ± 0.5) revealed the maximum antifungal activity against Candida albicans compared to Clotrimazole (13.0 ± 1.0 mm). Data regarding the antifungal activity of the

above mentioned plants are scanty. However, these results are promising in order to introduce new antifungal agents. Undoubtedly, the antimicrobial agents derived from such plants may eradicate bacteria or fungi by a mechanism different than that occurred by the synthetic or semi-synthetic antibiotics (Eloff, 1997), which could be much effective with less side effects. CONCLUSION The results of the present study provide support to the claim on these plants in Saudi folk medicine against some of the mentioned disorders and diseases. Plants of interest are those which exhibited antimicrobial activity higher than the antibiotics tested. These plants (Rhazya stricta, Francoeuria crispa and Blepharis ciliaris) should be subjected to intensive studies such as fractionation, isolation of the active constituents, toxicity against animal and human cells and so forth. ACKNOWLEDGEMENTS This study was supported by the Deanship of scientific research, Qassim University, Saudi Arabia, grant No. SR-D-1105. Authors are greatly thankful for this support.

REFERENCES Abdallah EM, Khalid AS, Ibrahim N (2009). Antibacterial activity of oleo-gum resins of Commiphora molmol and Boswellia papyrifera against methicillin resistant Staphylococcus aureus (MRSA). Sci. Res. Essay. 4 (4): 351– 356. Abdallah EM (2011). Plants: An alternative source for antimicrobials. J. Applied Pharma. Sci., 1(6): 16–20.

Abdallah EM, Hsouna AB, Al-Khalifa KS (2012). Antimicrobial, antioxidant and phytochemical investigation of Balanites aegyptiaca (L.) Del. edible fruit from Sudan. Afri. J. Biotech., 11(52): 11535–11542. Ahmad S, Fatima K, Atiq-ur-Rahman (2004). Antibacterial activity of Pakistani Rhazya stricta. Pakistan J. Sci. Indust. Res., 47 (1): 29–33.

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Al-Essa MA, Al-Mehaidib A, Al-Gain S (1998). Parental awareness of liver disease among children in Saudi Arabia. Ann. Saudi Med., 18 (1): 79–81. Alfarhan A H, Chaudhary SA, Thomas J (1998). Notes on the flora of Saudi Arabia (Third edition). J. King Saud Univ., 10(1): 31–40. Das K, Tiwari R KS, Shrivastava D K (2010). Techniques for evaluation of medicinal plant products as antimicrobial agent: Current methods and future trends. J. Med. Plants Res., 4(2): 104–111. Deeni YY, Sadiq NM (2002). Antimicrobial properties and phytochemical constituents of the leaves of African mistletoe (Tapinanthus dodoneifolius (DC) Danser) (Loranthaceae): an ethnomedicinal plant of Hausaland, North Nigeria J. of Ethnopharma., 83: 235–240. Edeoga H O, Okwu D E, Mbaebie B O (2005). Phytochemical constituents of some Nigerian medicinal Plants. African J. Biotech., 4 (7): 685–688. El-Ghazali GE, Al-Khalifa KS, Saleem GA, Abdallah EM (2010). Traditional medicinal plants indigenous to Al-Rass province, Saudi Arabia. J. Med. Plants Res., 4(24): 2680–2683. El-Kamali H H, Mahjoub SA (2009). Antibacterial activity of Francoeuria crispa, Pulicaria undulates, Ziziphus spina-christi and Cucurbita pepo anainst seven standard pathogenic bacteria. Ethnobotanical Leaflets, 13: 722–733. Eloff JN (1997). Which extract should be used for the screening and isolation of antimicrobial components from plants?. J. Ethnopharma., 60: 1–8.

El-Shanawany MA, Sayed HM, Ibrahim SRM, Fayed MMA, Radwan MM, Ross SA (2012). A new isoflavone from Blepharis ciliaris of Egyptian origin. Med. Chem. Res.DOI 10.1007/S00044012-0228-2. Krishnaiha D, Devi T, Bono A, Sarbatly R (2009). Studies on phytochemical constituents of six Malaysian medicinal plants. J. Med. Plants Res., 3(2): 67– 72. Kumar

A, Schweizer HP (2005). Bacterial resistance to antibiotics: Active efflux and reduced uptake. Advanced Drug Delivery Rev., 57: 1486–1513.

Rίos J L, Recio M C (2005). Medicinal plants and antimicrobial activity. J. Ethnopharma., 100: 80–84. Rohit S, Thakur G S, Sanodiya B S, Mukeshwar P, Prakash B (2012). Saponin: a wonder drug from Chlorophytum species. Global J Res. Med. Plants & Indigen. Med.,1(10): 503–515. Runyoro D, Matee M, Olipa N, Joseph C, Mbwambo H (2006), Screening of Tanzanian medicinal plants for antiCandida activity. BMC Complement Altern. Med., 6(11). doi:10.1186/14726882-6-11. Samie A, Obi C L, Bessong PO, Namrita L (2005). Activity profiles of fourteen selected medicinal plants from Rural Venda communities in South Africa against fifteen clinical bacterial species. African J. Biotech., 4 (12): 1443–1451. Sashikala GD, Kottai AM, Satheesh DK, Rekha S, Indhumathy, Nandhini R. (2009). Studies on the antibacterial and antifungal activities of the ethanolic extracts of Luffa cylindrica (Linn) fruit. Int. J. Drug Dev. Res., 1(1):105–109.

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 189–197

Shaw WV (1984). Bacterial resistance to Chloramphenicol, British Med. Bull., 40 (1): 36–41. Sofowora A (1984). Medical Plants and Traditional Medicine in Africa. (eds.) John Willey and Sons Ltd., Ibadan, Nigeria.

Source of Support: Qassim University,

Treadway S (1998). Exploring the Universe of Ayurvedic botanicals to manage bacterial infections, Clinical Nutrition Insights, 6 (17): 1–5. Watt E, Pretorius J C (2001). Purification and identification of active antibacterial components in Carpobotus edulis L., J. Ethnopharma., 76: 87–91.

Conflict of Interest: None Declared

Saudi Arabia

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Research article DIVERSITY AND AVAILABILITY STATUS OF ETHNO-MEDICINAL PLANTS IN THE LOHBA RANGE OF KEDARNATH FOREST DIVISION (KFD), GARHWAL HIMALAYA Ballabha Radha1*, Singh Dinesh2, Tiwari J K3, Tiwari P4 1, 2, 3, 4

Department of Botany and Microbiology, HNB Garhwal University, Srinagar Garhwal-246 174, Uttarakhand, India *Corresponding Author: E-mail: radhekuniyal.2007@rediffmail.com

Received: 20/02/2013; Revised: 25/02/2013; Accepted: 27/03/2013

ABSTRACT The present study has been carried out in the Lohba range of the Kedarnath Forest Division, Garhwal Himalaya to document the diversity, ethno-medicinal uses and availability status of medicinal plants. The inhabitants of the region are dependent up to a large extent on wild resources for their therapeutic needs. The region is rich in ethnomedicinal plant diversity. A total of 140 species belonging to 126 genera and 64 families were recorded from the study area. Out of the documented species 69 were herbs, 37 shrubs, 23 trees and the rest 11 were climbers. Out of the recorded plant species, 17 were abundant, 83 common and 40 uncommon to this area. Plant parts are used to cure cold, cough, fever, stomach disorders, joints pain, eye diseases, healing of cuts and wounds, toothache, etc. This study will be helpful in developing a comprehensive data base on the medicinal plant resources to strengthen the health care system in the area and in conserving the traditional knowledge for the prosperity of the remote village areas. KEYWORDS: Ethnomedicinal plants, availability status, Kedarnath Forest Division, Garhwal Himalaya.

Cite this article: Ballabha R, Singh D, Tiwari J K, Tiwari P (2013), DIVERSITY AND AVAILABILITY STATUS OF ETHNO-MEDICINAL PLANTS IN THE LOHBA RANGE OF KEDARNATH FOREST DIVISION (KFD), GARHWAL HIMALAYA, Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 198–212

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INTRODUCTION Ethnobotanical studies typically focus on recording the knowledge of traditional societies in remote places (Hodges and Bennett, 2006). In the remote areas traditional customs and beliefs are still maintained and modern trends are yet to reach, which provide interesting scope of ethnobotanical studies (Tiwari et al., 2010a). Indigenous people of different parts of the world have a vast knowledge of, and capacity for, developing innovative practices and products from their environment. Indigenous knowledge grows from close interdependence between knowledge, land, environment and other aspects of culture in indigenous societies, and the oral transmission of knowledge in accordance with well understood cultural principles and rules regarding secrecy and sacredness that govern the management of knowledge (Tripathi et al., 2000). According to WHO approximately 80% of world population in developing countries depends on traditional medicines for primary healthcare (WHO, 2002) and in modern medicine too, nearly 25% are based on plant derived drugs (Tripathi, 2002). Garhwal Himalaya occupies an important place in Indian subcontinent and has its peculiar topography, vegetation, people and traditions. About 80% of the total population is rural and the inhabitants are called the Garhwalis or Paharis. They have their own cultures, medicines, foods, etc. and are well versed with valuable knowledge accumulated through a long period of experience. Even now they are dependent on the natural resources from the forests for their sustenance and for the treatment of various ailments (Tiwari et al., 2010a). The plants are still serving as remedies for various ailments in crude form, as modern medicine has not adequately armed the therapeutic arsenal of the natives of remote areas. The literature abounds in investigations on folk medicines in different parts of Garhwal Himalaya (Gaur et al., 1984, 1985, 1987; Tiwari, 1986; Negi et al., 1993; Maikhuri et al.,

1998; Gaur, 1999; Badoni and Badoni, 2001; Negi et al., 2002; Semwal et al., 2010; Tiwari et al., 2010b) but little attention has been paid on plants used in ethonomedicine from the Lohba range of the Kedarnath Forest Division (KFD). Documentation of such practices is required in view of gradual disappearance of this knowledge in new generations. Therefore, an attempt has been made to record the diversity and indigenous uses of plants in ethnomedicine in the Lohba range of KFD.

MATERIALS AND METHODS Study Area KFD is situated in the north-west part of the Himalaya and well known for its rich biodiversity. The inhabitants of the area largely depend on plants for food, fodder, medicine, timber, fuel-wood, dye, beverage, and various religious and cultural needs. Geographically, the area stretches between 29° 57' 33" to 30° 06' 05" N latitudes and 79°11' 33" to 79° 20' 33" E longitudes with the altitude ranging from 1268 m to 3067 m asl (Fig. 1). The total geographical area of region is about 16387.40 ha which represents 26.76 % of the Division. Western Ramganga is the main river of this area, which originates from the lesser Himalayan mountain range (Dhudhatauli) and enters into Corbett National Park after flowing 100 km with its tributaries. Besides providing perennial water source it provides habitat to many plant and animal communities. The mountainous tract of the whole region is varying in altitude which contributes variation in the climatic conditions to play an important role in the distribution of the vegetation in the area. Summer, rainy and winter seasons are well marked due to fluctuating precipitation, temperature, light, wind, humidity and even day- length. The maximum temperature was recorded during the months of May and June (25–30°C), whereas minimum in the months of December and January (10–20°C). Rainfall, snowfall, hailstorm, dew, frost, etc, are the main form of precipitation and the average annual rainfall

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was 110 mm in the region. The average annual relative humidity was 42% which decreases with increase in temperature and altitude in study area. The maximum humidity reaches near to 80–92% during July and August, whereas the minimum humidity (25–38%) was recorded in the months of January and December.

The vegetation of the Lohba Range is characterized by sub montane and montane zone types. The area is represented by Pinemixed forest (1200–1500 m), Oak forest (1800–2000 m), Oak-mixed forest (1500– 2500 m) and Oak-Abies mixed forest (2700– 3114 m) however, some patches are occupied by pine and scrub forest along with grassy slopes.

Fig. 1 Map showing the study area.

Fig. 2 Plant parts being preferred for medicine by local inhabitants. 60 50

Number of species

40 30 20 10 0

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Methodology

RESULTS AND DISCUSSION

Extensive field surveys were made in the study area during the years 2009–2011 for the survey of the vegetation and ethno-medicinal uses. A structured questionnaire was used to collect data on local name of plants, uses, parts used and mode of application. Ethno-medicinal information on plants was collected through interviewing local communities. The informants were medicine-men (Vaidhyas), peasants, shepherds, priests and village headmen. To determine the authenticity of information collected during field work, repeated verification of data from different informants was done. Thus, only the specific and reliable information, cross-checked with informants has been incorporated in the present study. Recorded plant species were identified with the help of Garhwal University Herbarium (GUH), Herbarium of the Botanical Survey of India, Northern Circle, Dehradun (BSD) and regional Floras (Duthie, 1906; Osmaston, 1927; Rau, 1961; Naithani, 1984-85; Gaur, 1999). The availability status of plants such as abundant, common and uncommon was given based on their occurrence in the study area.

The study revealed 140 medicinal plant species belonging to 64 families in the Lohba range of the Kedarnath Forest Division, Garhwal Himalaya. The availability status and ethno-medicinal uses of the plant species are presented in Table 1. The recorded species diversity represents trees (23 species), shrubs (37), herbs (69) and climbers (11). The families, Asteraceae (15 species), Lamiaceae (10), Rosaceae (10), Solanaceae (5), Liliaceae (4), Orchidaceae (4), Ranunculaceae (4), Rutaceae (4), Acanthaceae (3), Apiaceae (3), Caesalpiniaceae (3), Cucurbitaceae (3), Euphorbiaceae (3), Rubiaceae (3) and Zingiberaceae (3) were major representations, whereas Artemisia (3 species), Cassia (3), Anaphalis (3), Asparagus (2), Berberis (2), Ficus (2), Geranium (2), Mentha (2), Polygonatum (2), Rumex (2), Salvia (2) and Swertia (2) were the genera with more than one species being used (Table 1). As per the plant parts, root/rhizome of 54 species was used to cure different ailments (Fig. 2), followed by leaves (53 species) and whole plant (30 species). Plant parts are being used to cure cold, cough, fever, stomach disorders, joints pain, eye diseases, healing of cuts and wounds, toothache etc.

Table 1. Diversity, availability status and ethno-medicinal uses of plant species in the Lohba range of Kedarnath Forest Division (KFD), Garhwal Himalaya. S. No.

Botanical Name

Local Name

Family

Elevation (m asl)

Life Form1

Availability

Abies pindrow Royle

Raisul

Pinaceae

2500–3000

Bark extract is given in cough and bronchitis.

Achyranthes aspera L.

Lich kura

Amaranthaceae

1300–2300

Aconitum balfourii Staf

Meetha jari

Ranunculaceae

2900–3000

Acorus calamus L.

Bauj

Araceae

1600–2200

Root infusion is given in fever. Leaf extract is given to women to facilitate delivery. Plant decoction is given in dropsy and bronchitis. Root paste is used in snakebite and also applied over chest in chest pain. Extract of rhizome is given in gastric troubles.

Status2

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Ethnomedicinal uses

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Adhatoda zeylanica Medikus

Basinga

Acanthaceae

1200–1400

Aesculus indica (Colebr. ex Cambess) Hook. Ageratum conyzoides L.

Pangar

Hippocastanaceae

2000–2500

Gunrya

Asteraceae

1300–2000

Kuriya

Rosaceae

1200–2500

Neelkanthi

Lamiaceae

1200–1500

Leaf extract is given in fever. Plant extract used as a tonic.

Jonkmari

Primulaceae

1200–2500

Externally applied in dropsy.

Anaphalis adnata Wallich ex DC. Anaphalis busua (Buch.Ham. ex D.Don) DC. Angelica glauca Edgev.

Kabash, Bugla

Asteraceae

1300–2500

Leaf paste is applied on cuts and wounds.

Bugla, Buglya

Asteraceae

1600–2200

Leaf juice is applied on bruises, wounds and cuts.

Choru

Apiaceae

2900–3000

Root is given with tea in cough and cold.

Arisaema tortuosum (Wallich) Schott. Artemisia capillaris Thunb.

Bagmungari

Araceae

1600–2200

Paste of tuber is applied on the burns and cuts.

Jhirun

Asteraceae

1300–2300

Stem and leaf juice is given in fever and constipation. Root power used in stomachache Plant juice is given in intestinal worms and externally applied on cuts and wounds. Plant extract is given in fever and also used in skin diseases.

10 11

13 14

Agrimonia pilosa Ledebour Ajuga bracteosa Wallich ex Benth. Anagallis arvensis L.

Root bark is given in fever. Young twigs used for cough and cold. Leaf juice with honey is given in cough and fever. Root juice is given in bowl complaints.

Root paste is applied on sores, cuts and various skin diseases. Plant decoction is given in cough and diarrhea.

Artemisia nilagrica (Clarke) Pamp.

Kurnja

Asteraceae

1600–2200

Artemisia roxburghiana Wallich ex Besser Asclepias curassavica L.

Kurnja

Asteraceae

1600–2200

----

Asclepiadaceae

1300–1500

Latex applied on cuts and wounds.

Asparagus adscendens Buch.-Ham. ex Roxb. Asparagus racemosus Willd

Jhirni

Liliaceae

1500–2200

Tuberous roots with honey are used in dysentery.

Jhirni

Liliaceae

1300–1500

Roots extract is given in fever.

18 19

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Barleria cristata L.

Kalabansa

Acanthaceae

1200–2300

Leaves are crushed, mixed with seeds of black pepper and given orally in dyspepsia.

Berberis aristata DC.

Kirmor

Berberidaceae

1700–3000

+++

Stem and root juice is applies in ophthalmic infections.

Berberis asiatica Roxb. ex DC.

Kirmor

Berberidaceae

1200–2500

Stem bark and root juice used in eye afflictions. Infusion of root given in fever.

Bergenia ciliata (Haworth) Sternberg Bidens pilosa L.

Silpari

Saxifragaceae

2200–2400

+++

Root paste is given in fever, urinary and renal troubles.

Kumra

Asteraceae

1300–2700

Vegetable of the plant useful in skin ailments.

Genthi

Urticaceae

1200–1400

Paste of bark is applied on the bone fracture.

Upaniy a-Ghas

Rutaceae

1200–1700

Leaf paste is applied on cuts and wounds.

Pundera

Nyctaginaceae

1200–1700

Brugmansia suaveolens (Humb. & Bonpl. ex Willd.) Berchtold & J. S. Presl Buddleja asiatica Lour.

Dhatura

Solanaceae

1200–1400

Roots chewed as tonic. Leaf extract used in eye diseases. Plant infusion is given in asthma and bronchitis. Leaf, flower and seed paste is applied on joints pain.

----

Buddlejaceae

1300–1600

Leaf juice is applied on skin eruption.

Callicarpa macrophylla Vahl. Cannabis sativa L.

Daiya

Verbenaceae

1200–1500

Stem bark is used in skin ailments.

Bhang

Cannabaceae

1300–2100

+++

Leaves boiled with butter are taken in vomiting. Leaf juice poured in ear in ear pain. Leaves are used as an intoxicating agent.

Cassia occidentalis L.

Taror

Caesalpiniaceae

1200–1400

Cassia tora L.

Chakuda

Caesalpiniaceae

1200–1400

Leaves used in skin disease. Decoction of roots is given in dropsy. Leaf and root paste useful in piles and ringworms. Leaves and seeds are used in skin diseases, cuts, wounds and bone fracture.

25 26

30 31

Boehmeria regulosa Wedd. Boennighause nia albiflora (Hook.) Reichb ex Meisn. Boerhavia diffusa L.

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Cassia fistula L.

Amaltas

Caesalpiniaceae

1200–1400

Cedrus deodara (Roxb. ex D.Don) G.Don

Deodar

Pinaceae

1400–2200

Centella asiatica (L.) Urban Cinnamomum tamala (Buch.Ham.) Nees & Ebermaeir Cissampelos pareira L.

Brahmi

Apiaceae

1400–2400

Leaves paste is applied externally in skin diseases.

Dalchini

Lauraceae

1300–1700

Leaves paste is used in throat irritation.

Parha

Menispermaceae

1200–3000

Citrus aurantifolia (Christmann) Swingle Clematis buchananiana DC. Coccinia grandis (L.) Voigt Colebrookia oppositifolia J.E. Smith

Kagzinimbu

Rutaceae

1300–2200

Root juice is given to children in dyspepsia, diarrhea, stomach ache and colic. Leaf decoction inhaled in headache, cold and fever.

Laguliya

Ranunculaceae

1200–3000

Decoction of leaves and roots is applied in scabies.

Kandaroi

Cucurbitaceae

1200–2000

Binda

Lamiaceae

1300–2000

Root paste is applied on the pelvic region in suppressed urination. Leaf paste applied on cuts and wounds.

Bani

Rosaceae

1300–2800

Root paste is applied on cuts and wounds.

Haldi

Zingiberaceae

1400–2100

Rhizome paste is applied on cuts and wounds. It is also used as antiseptics.

Cotoneaster microphyllus Wallich ex Lindley Curcuma longa L.

Fruits are given to women to expel the placenta. Fruit pulp is given in asthma, bronchitis and skin diseases. Paste of bark is externally applied on piles and arthritis.

Cuscuta europaea L.

Akaslaguli

Cuscutaceae

1600–2300

+++

Plant extract used in skin diseases.

Cynodon dactylon (L.) Persoon Cynoglossum glochidiatom Wallich ex Benth. Datura metel L.

Doob

Poaceae

1300–3000

Roots are taken in fever and internal injury.

Likhkura

Boraginaceae

1500–2000

+++

Root paste is applied on sores.

Dhatura

Solanaceae

1200–1400

Leaf and Seed paste is applied on sores and used as a massage in arthritis.

Desmodiun elegans DC.

Chamliya

Fabaceae

1500–2700

Root decoction is given in renal disorder.

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Kuthi

Acanthaceae

1300–2600

+++

Leaf paste applied on wounds, check bleeding.

Geithi

Dioscoreaceae

1400–1900

Duchesnea indica (Andrews) Focke Echinops cornigerus DC. Eupatorium adenophorum Sprengel Euphorbia royleana Boissier Ficus palmata Forsk.

Kaphliya

Rosaceae

1400–2200

Root and fruit paste is applied on burns and wounds. Leaf juice is given in diarrhea.

Kandara

Asteraceae

1200–2000

Root juice is taken in urinary trouble and fever.

Guyajhar

Asteraceae

1300–3000

Crushed leaves applied on cuts and wounds.

Sulla

Euphorbiaceae

1800–2000

Latex is used as an antiseptic on cuts and wounds.

Bedu

Moraceae

1300–2000

Ficus religiosa L.

Peepal

Moraceae

1300–1500

Foeniculum vulgare (L.) Miller

Saup

Apiaceae

1700–2000

Fruits are used in digestive disorders. Latex is applied on the pimples. Root bark, young shoots and fruit decoction is given in sexual weakness in men. Infusion of bark is given in constipation. Seed paste is applied on mouth during the teething of child.

Fumaria indica (Haussknecht) Pugsley Geranium nepalense Sweet

Kherua

Fumariaceae

1300–2200

Plant decoction is given in fever and suppressed urination.

----

Geraniaceae

1600–2200

Ratanjot

Geraniaceae

2200–3000

Roots are used as an antiseptic. Root extract is given in lever troubles and fever. Root paste is applied on wounds and cuts.

Bhaiska nali

Urticaceae

1500–3000

Leaf juice is given in gonorrhea.

Bukhil

Asteraceae

1300–2500

Juice of stems and leaves is given in bruises and cuts.

Banhaldi

Zingiberaceae

2000–2300

Rhizome powder is used in asthma.

Papri

Ulmaceae

1700–1900

Bark decoction is used in rheumatic pain.

Geranium wallichianum D. Don ex Sweet Girardinia diversifolia (Link.) Friis. Gnaphalium hypoleucum DC. Hedychium spicatum Buch.-Ham. ex J. E. Smith Holoptelea integrifolia (Roxb.) Planchon

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Phioli

Hypericaceae

1500–2000

----

Asteraceae

1300–2500

Sulkairi

Convolvulaceae

1300–2300

Jasminum humile L.

----

Oleaceae

1200–2500

Juglans regia L.

Akhor

Juglandaceae

1300–2300

Leucas lanata Benth.

Niras

Lamiaceae

1300–2600

Lyonia ovalifolia (Wallich) Drude Mallotus philippensis (Lam.) Muell.Arg.

Anyar

Ericaceae

1400–3000

+++

Leaves juice is applied in eczema.

Ruina

Euphorbiaceae

1200–1300

Malva verticillata L.

----

Malvaceae

1400–2000

Bark juice is given to children in diarrhea and dysentery. Paste of fruit powder is applied externally on cuts, wounds. Root and seed paste is applied on sores and skin eruptions. Root paste is given in whooping cough.

Martynia annua L.

Bichu

Martyniaceae

1200–1800

Fruit powder is given in cold and cough.

Melia azedarach L.

----

Meliaceae

1200–1400

Mentha arvensis L.

Paudina

Lamiaceae

1200–2300

Decoction of bark used in gonorrhea, bark paste is applied on skin eruptions. Infusion of heart wood is given in asthma. Leaves, fruits and seeds are useful in skin diseases Plant extract used in vomiting and indigestion.

Mentha piperita L.

Pepermint

Lamiaceae

1200–2300

Plant extract used in indigestion.

Micromeria biflora Buch.Ham. ex D.Don Mukia maderaspatan a (L.) M. Roemer Murraya koenigii (L.) Sprengel

Ban ajwain

Lamiaceae

1300–3200

+++

GuliyaKakri

Cucurbitaceae

1300–2100

Crushed leaves inhaled in cold and sinusitis. Leaf extract with milk is given in gastroenteritis. Fruit paste is used in urinary disorder and vomiting.

Karipatta

Rutaceae

1200–1400

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Plant juice is given in diarrhea and intestinal worms. Root juice is given in urinary trouble.

Decoction of seed is given in fever and constipation. Seed paste is applied in urticaria. Root juice is given to children in stomach troubles. Leaves used as fungicide and insecticide and bark used in bone fractures. Plant extract is given in cough.

Bark, leaves and roots are used as insecticide.

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85 86

Nasturtium ---officinale R.Br. Nicotiana Bantamb plumbaginifoli aku a Viviani Tambaku Nicotiana rustica L.

Brassicaceae

1200–2500

Plant juice is given in diarrhea.

Solanaceae

1200–1400

Leaf juice is applied on sores.

Solanaceae

2000–3000

Leaves juice is applied on cuts and wounds.

Oberonia falconeri Hook.f. Ocimum tenuiflorum L.

----

Orchidaceae

2000–2300

Plant extract is given in diarrhea and bronchitis.

Tulsi

Lamiaceae

1200–1600

Plant used in fever, cold, cough, colitis, urinary troubles and vomiting. Milky juice obtained from immature capsules is given to children in fever, dysentery and cholera. Plant extract is used to stimulate vomiting.

Papaver somniferum L.

Post

Papaveraceae

2900–3000

Parnassia nubicola Wallich ex Royle

Phutkya

Saxifragaceae

2200–2400

Peperomia tetraphylla (Forster f.) Hook. & Arn. Perilla frutescens (L.) Britton

Tirpirya

Piperaceae

1500–3000

+++

Leaf paste is applied on wounds and burns.

Bhangje era

Lamiaceae

1500–2100

Plant extract or power of dried plant parts used for cold, cough, bronchitis and uterine ailments; leaf paste applied on arthritis. Fruits paste is used against wounds and sores.

Phoebe lanceolata (Nees) Nees

Kaula

Lauraceae

1400–1700

Pholidota articulata Lindley Phyallanthus emblica L.

Jewanti

Orchidaceae

2000–2300

Plant extract is used as tonic.

Aola

Euphorbiaceae

1200–1400

Polygonatum multiflorum (L.) Allioni

----

Liliaceae

1200–1400

Fruits are used in digestive disorders and fruit juice useful in leucorrhoea. Fruit powder is given in fever Decoction of rhizome is given in urinal disorders

Polygonatum verticillatum (L.) Allioni Populus ciliata Wallich ex Royle Potentilla fulgens Wallich ex Hook. Prinsepia utilis Royle

----

Liliaceae

1400–2300

Root paste is applied on wounds.

Pupular

Salicaceae

1300–2600

Leaf juice is used as blood purifier and stimulant.

Bajrdanti

Rosaceae

2300–3000

Plant juice is applied on mouth in tooth ache.

Bhainkal

Rosaceae

1300–3000

+++

Seed oil is applied in joint pain and arthritis.

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101

Prunus cornuta (Wallich ex Royle) Steudel Punica granatum L.

Jamnoi

Rosaceae

2400–3000

Bark decoction is given in diarrhea.

Darim

Punicaceae

1300–1700

Fruit juice is used in cough and diarrhea. Ripened fruits are used in digestive disorders. The fruit is crushed with teeth and juice is forced into the eyes of cattle in cataract and injuries. Leaves paste is applied in skin diseases.

102

Pyrus pashia Buch.-Ham. ex D.Don

Melu

Rosaceae

1200–2900

+++

103

Ranunculus arvensis L.

Chambul

Ranunculaceae

1200–3000

+++

104

Raphanus sativus L.

Muli

Brassicaceae

1200–2500

Cooked leave and leave juice is given in jaundice.

105

Reinwardtia indica Dumortier Rhododendron arboreum Smith Rosa brunonii Lindley

Phuenli

Linaceae

1200–2200

Leaf juice is applied on cuts and wounds.

Burans

Ericaceae

1200–3000

+++

Kunja

Rosaceae

1400–2000

Flower juice is given in hart troubles and diseases. Young shoot poisonous to cattle. Leaf juice used in cuts, wounds. Dried flower powder used in diarrhea

108

Rosa sericea Lindley

Dhurku nja

Rosaceae

2700–3000

Flower juice is applied externally in eye diseases.

109

Rubia manjith Roxb. ex Fleming

Majeithi

Rubiaceae

1300–3000

+++

110

Rubus niveus Thunb.

Kali Hinsar

Rosaceae

1200–2200

111

Rumex hastatus D.Don

Almor

Polygonaceae

1300–2300

+++

Stem used as an antidote to snake bite; roots are used as a tonic; flower extract used in bacillary dysentery. Root decoction is given in diarrhea. Juice of young shoots and roots is given in stomachache. Leaf extract applied on cuts and wounds to check bleeding.

112

Rumex nepalensis Sprengel

Khoya

Polygonaceae

1300–3000

Leaf juice is given in digestive disorders.

113

Salvia lanata Roxb.

Pakhuliya

Lamiaceae

1300–3000

Flower paste is given in cold and cough.

114

Salvia nubicola Wallich ex Sweet Sarcococca saligna (D.Don) Muell.-Arg. Satyrium nepalens D.Don

Ganiya

Lamiaceae

2500–2700

Root paste is given in cold and cough.

Peruli

Buxaceae

1300–3000

Leaf paste is applied on joint pain.

----

Orchidaceae

1600–2100

Plant extract is used in diarrhea.

106

107

115

116

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Sida rhombifolia L.

Bhiunli

Malvaceae

1200–2100

Leaves and root bark used in gonorrhea.

118

Smilax aspera L.

Kukrdar

Smilacaceae

1400–2400

+++

119

Solanum nigrum L.

Makoi

Solanaceae

1200–2300

120

Solidago virgaurea L.

----

Asteraceae

1800–3000

Root paste mixed with mustard oil is applied in arthritis and joint pain. Plant extract is given in liver trouble, piles and dysentery. Fruits are useful in diarrhea. Plant juice is applied on cuts and in scabies.

121

Sonchus asper (L.) Hill

Dudhi

Asteraceae

1300–2200

Plant juice is used as blood purifier.

122

Spermadictyon sauveolens Roxb. Stellaria media (L.) Villars Stephania glabra (Roxb.) Miers Swertia angustifolia Buch.-Ham. ex D.Don Swertia chirayita (Roxb. ex Fleming) Karsten Symplocos paniculata (Thunb.) Miq. Tanacetum dolichophyllu m (Kitamura) Kitamura Taraxacum officinale Weber Taxus baccata L.

Padera

Rubiaceae

1400–2600

+++

Leaf juice is applied on cuts, wounds and sores.

Badyalu

Caryophyllaceae

1200–1700

Gindaru

Menispermaceae

2000–2500

Plant paste is externally applied on burns, wounds and boils. Root juice is given in fever and dysentery.

Chirota

Gentianaceae

2100–3000

Seed powder is given in cough and asthma.

Chirota

Gentianaceae

2100–3000

Seed powder is given in cough and asthma.

Lodh

Symplocaceae

1500–2500

+++

Decoction of bark is given in diarrhea.

Dhoop

Asteraceae

2800–3000

Plant juice is given to children to expel intestinal worms.

Kanphu liya

Asteraceae

1300–2000

Latex is applied over corns and warts.

Thuner

Taxaceae

2500–3000

Bark paste is applied in headache and bone fracture. Root powder mixed with honey is given in fever, dyspepsia and piles. Root juice is given as emetic.

123

124

125

126

127

128

129

130

131

Thalictrum foliolosum DC.

Mamira

Ranunculaceae

1200–2200

132

Trichosanthus tricuspidata Lour. Valeriana wallichii DC.

Indraiyan

Cucurbitaceae

1400–2300

Sumaya

Valerianaceae

1400–2600

Root paste is applied in muscular pain.

----

Orchidaceae

1600–2100

Plant paste is applied in bone fractures.

133

134

Vanda cristata Lindley

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Verbascum thapsus L.

----

Scrophulariaceae

1200–2500

136

Viburnum grandiflorum Wallich ex DC. Viola pilosa Blume.

Ghenu

Caprifoliaceae

2800–3000

Kaura

Violaceae

1200–2100

137

Plant extract used in bronchitis and asthma. Seeds are used as sedative. Bark decoction is used in hepatic trouble.

Decoction of plant useful in fever and bronchitis. Root is used as an emetic. Leaf juice is applied on cuts and wounds. Leaves, roots and fruits pastes are applied in arthritis.

138

Vitex negundo L.

Siwain

Verbenaceae

1300–1800

139

Zanthoxylum armatum DC.

Timru

Rutaceae

1400–1800

Leaves and fruits chewed for mouth wash and tooth care. Seed paste is applied on teeth in toothache.

140

Zingiber officinale Roscoe

Adu

Zingiberaceae

1300–1900

Used in cold, cough and stomach troubles.

Abbreviations used: 1. H = herb, S = shrub, T = tree, Cl = climber, 2. +++ = Abundant, ++ = Common, + = Uncommon The present study indicates that the area harbors a high diversity of medicinal plants. Out of 140 plant species, 17 were abundant, 83 common and 40 uncommon to this area. Species like Abies pindrow, Aconitum balfourii, Adhatoda zeylanica, Aesculus indica, Angelica glauca, Artemisia roxburghiana, Boehmeria regulosa, Boerhavia diffusa, Brugmansia suaveolens, Buddleja asiatica, Centella asiatica, Foeniculum vulgare, Geranium nepalense, Geranium wallichiana, Malva verticillata, Martynia annua, Melia azedarach, Murraya koenigii, Nicotiana plumbaginifolia, Oberonia falconeri, Ocimum tenuiflorum, Perilla frutescens, Pholidota articulata, Polygonatum multiflorum, Polygonatum verticillatum, Punica granatum, Rosa sericea, Satyrium nepalens, Sida rhombifolia, Stephania glabra, Swertia chirayita, Taxus baccata, Vanda cristata, Vitex negundo etc. are uncommon to this area and being threatened due to unplanned exploitation. The inhabitants revealed abundance of many of these species in the past, which has got restricted now to certain patches. If immediate steps for their sustainable utilization and conservation are not taken, these

species may reach to the status of threatened in the area. CONCLUSION It is evident from the investigation that the local people have great familiarity with the plants of their ambient environment which has immense importance in advancement of modern sustainable technology. The occurrence of a number of economically important medicinal plant species demands the conservation of these species as the day-to-day need of forest resources particularly for medicine has increased the pressure in the area and may lead to reduction of these species. Therefore, there is a need to develop adequate strategy and action plan for the conservation and management of habitats and species. ACKNOWLEDGEMENTS The authors are grateful to the inhabitants of the Lohba range of the Kedarnath Forest Division (KFD) for providing the information about the ethno-medicinal uses of the plant resources.

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REFERENCES Badoni AK, Badoni K (2001). Ethnobotanical heritage. In: Kandari OP, Gusain OP (eds) Garhwal Himalaya: Nature, Culture and Society Transmedia, Media House, Srinagar, Garhwal, pp 127–147. Duthie JF (1906). Catalogue of plants of Kumaon and of the adjacent portions of Garhwal and Tibet based on the collections made by Strachey and Winterbottom during the years 1846– 1849. London. Reprint 1994, Bishan Singh Mahendra Pal Singh, Dehradun. Gaur RD (1999). Flora of the District Garhwal, North West Himalaya (With Ethnobotanical Notes). Trasmedia: Srinagar (Garhwal), Uttarakhand, India. Gaur RD, Semwal JK, Tiwari JK (1984). A survey of high altitude medicinal plants of Garhwal Himalaya. Bull. Medic. Ethnobot. Res., 3: 102–116. Gaur RD, Tiwari JK (1987). Some little known medicinal plants of Garhwal Himalaya: An ethnobotanical study. In: Leeuwenberg AGM (ed). Medicinal and Poisonous Plants of the Tropics Pudoc, Wageningen, Netherlands, pp 139–142. Gaur RD, Tiwari JK, Negi KS (1985). Plants used for magico-religious practices by tribals of Garhwal Himalayas. Proceedings of 2nd Annual Workshop on MAB Projects. Dept. of Environment, New Delhi, pp 88–95. Hodges S, Bennett BC (2006). The ethnobotany of Pluchea carolinesis (Jacq.) G. Don (Asteraceae) in the Botanicas of Miami, Florida. Econ. Bot., 60(1): 5–84.

Maikhuri RK, Nauityal S, Rao KS, Saxena KG (1998). Role of medicinal plants in the traditional health care system: a case study from Nanda Devi Biosphere Reserve. Current Science, 75(2): 152– 157. Naithani BD (1984-85). Flora of Chamoli. 2 Vols. Botanical Survey of India, Howrah. Negi CS, Nautiyal S, Dasila L, Rao KS, Maihkuri RK (2002). Ethnomedicinal Plant Uses in a Small Tribal Community in a Part of Central Himalaya, India. J. Hum. Ecol., 14(1): 23–31. Negi KS, Tiwari JK, Gaur RD, Pant KC (1993). Notes on ethnobotany of five districts of Garhwal Himalaya, U.P., India. Ethnobot., 5: 73–81. Osmaston AE (1927). A Forest Flora for Kumaun. Government Press, Allahabad. Reprint 1990, Bishan Singh Mahendra Pal Singh, Dehradun. Rau M A (1961). Flowering plants and ferns of north Garhwal, Uttar Pradesh, India. Bull. Bot. Surv. India, 3: 215–251. Semwal DP, Kala CP, Bhatt AB (2010). Medicinal Plants and Traditional Health Care Knowledge of Vaidyas, Palsi and Others: a Case Study from Kedarnath Valley of Uttarakhand, India. Medicinal Plants, 2(1): 51–57. Tiwari JK (1986). Medicinal Plants of Garhwal Himalaya: An Ethnobotanical Survey. Unpublished D. Phil. Thesis University of Garhwal, Srinagar Garhwal (U.P.), India.

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Tiwari JK, Radha Ballabha, Tiwari P (2010a). Ethnopaediatrics in Garhwal Himalaya, Uttarakhand, India (Psychomedicine and Medicine). New York Science Journal, 3(4): 123–126.

Himalayan Medicinal Plants. In: Samant SS, Dhar U, Palni LMS (eds) Himalayan Medicinal Plants Potential and Prospects, Gyanodaya Prakashan, Nainital, pp 151–156.

Tiwari JK, Radha Ballabha, Tiwari P (2010b). Diversity and Present Status of Medicinal Plants in and around Srinagar Hydroelectric Power Project in Garhwal Himalaya, India: Needs for Conservation. Researcher, 2(2): 50–60.

Tripathi S, Varma S, Goldey P (2000). Using plants for health: indigenous knowledge in health care in a tribal region of Bihar, India. Int. J. Sust. Develop. & World Ecol., 7: 321–332.

Tripathi G (2002). Indigenous Knowledge and Traditional Practices of Some

Source of Support: Nil

WHO (2002). WHO Traditional Medicine Strategy 2002–2005. World Health Organization, Geneva.

Conflict of Interest: None Declared

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Research article ANTICATARACT ACTIVITY OF ERVATAMIA CORONARIA LEAF EXTRACT ON CHEMICALLY INDUCED CATARACTOGENESIS IN RATS Rathnakumar K1*, Jaikumar S2, Duraisami R3, Sengottuvelu S4 1, 2

Department of Ophthalmology, Sri Lakshminarayana Institute of Medical Sciences, Pondicherry, India Department of Pharmacology, Nandha College of Pharmacy and Research Institute, Erode- 638052 *Corresponding Author: Email: sehejan@gmail.com 3, 4

Received: 20/02/2013; Revised: 25/02/2013; Accepted: 27/03/2013

ABSTRACT The anticataract activity of the Ervatamia coronaria leaf extract was evaluated against naphthalene induced cataract in Wistar albino rats. Thirty rats in five groups were used for the study. Ervatamia coronaria leaf extract at dose levels (200 and 400 mg/kg) respectively and Vitamin E (50 mg/kg) were used as standard drugs while liquid paraffin was used for control. The test drugs were administered simultaneously with naphthalenefor 25 days. Naphthalene (0.5 g/kg for first 3 days and 1 g/kg thereafter for a period of 25 days) was used to induce cataract. The percentage of cataract incidence and opacity index were examined using ophthalmoscope. Naphthalene produced a marked mature cataract and an increase in the opacity index at various stages. The extract treated animals showed decrease in the onset and maturation of cataract against naphthalene challenge. From the results it was concluded that Ervatamia coronaria leaf extract protected the cataract maturation induced by naphthalene and it exhibited anticataract activity. KEYWORDS: Ervatamia coronaria, Cataract, Naphthalene

Cite this article: Rathnakumar K, Jaikumar S, Duraisami R and Sengottuvelu S (2013), ANTICATARACT ACTIVITY OF ERVATAMIA CORONARIA LEAF EXTRACT ON CHEMICALLY INDUCED CATARACTOGENESIS IN RATS, Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 213–218

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INTRODUCTION Cataract is opacity of the lens that interferes with vision, and is the most frequent cause of visual impairment worldwide, especially for the elderly, because the incidence of cataracts increases with increasing age. It is the leading cause of blindness and contributes to 50% of blindness worldwide (Haque and Gilani, 2005). The only present remedy for cataract is surgery. The opacity of lens occurs as a result of oxidation which is augmented by the free radical generation. The intensity of opacification of lens can be reduced by the antioxidants which scavenge the generation of free radicals. Previous studies confirmed that diet rich in vitamins, carotenoids and flavonoids may reduce the cataract intensity (Bunce et al., 1990). Normal levels of the antioxidantâ&#x20AC;&#x2122;s defense mechanism are not sufficient for the eradication of free radical induced injury. Therefore, the administration of antioxidants from a natural origin has a promising role to play. Several antioxidants of plant materials have been experimentally proven and widely used as more effective agents against oxidative stress. Ervatamia coronaria Stapf Local name : Adukkunandiyavattai (In Thamizh Language) (Synonym: Tabernaemontana divaricata) belongs to the family Apocynaceae, is a glabrous, evergreen tree indigenous to India and is cultivated in gardens for its ornamental and fragrant flowers. This species has been extensively investigated and a number of chemical constituents such as alkaloids (Pawelka and Stoeckigt., 1983), triterpenoids, steroids (Sharma and Cordell., 1988), flavonoids (Daniel and Sabnis., 1978) and phenolic acids (Henriques et al., 1996) were isolated from leaves, roots and stems of the plant. In Indian traditional system of medicine the plant material is widely used as a purgative, tonic to the brain, spleen and the liver (Kirtikar and Basu., 1975). Also used in the treatment of cancer, wounds and inflammations (Kirtikar and Basu., 1975). The plant extract was also

found to possess analgesic, antipyretic, vasodilator and CNS depressant effects (Taesotikul., 1989), antispasmodic, hypotensive activity (Dhar et al., 1968), anti-inflammatory (Henriques et al., 1996), uterine stimulant effect (Da Sil Va et al., 1984), cytotoxic (Yamamoto et al., 1997) and anti oxidant activity (Malaya Gupta et al., 2004). Traditionally the plant is also used in ophthalmic disorders. Hence the present study was aimed to evaluate the anti-cataract activity of the leaf extract of E. coronaria.

MATERIALS AND METHODS Drugs and chemicals Naphthalene and vitamin E were obtained from SD fine chemicals, Mumbai, India. All other drugs and chemicals used in the study were of analytical grade. Plant material Leaves of Ervatamia coronaria were collected from outskirts of of Erode, Tamilnadu. Authentication has been done by Prof. V. S. Kumar, Scientists (F) and Head of the Office, Tamilnadu Agriculture University, Coimbatore (Tamilnadu).The voucher specimen (No.: BSI/ SRC/ 5/ 23/ 12-13/ Tech. 816) has been deposited in the herbarium for future references. Preparation of extract The leaves were washed with fresh water to remove adhering dirt and foreign particles. The leaves were shade dried, crushed and grinded to get coarse powder. The coarse powder was then placed with 90% ethanolic solution in a round bottomed flask. 500 g of the coarse powder of the leaves of Ervatamia coronaria in 1.0 liter of 90% ethanolic solution were macerated for 7 days. The mensturm was collected, concentrated by vacuum distillation and then air dried in an evaporating dish till constant weight was obtained.

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Animals Wistar albino rats of either sex weighing 150–200 g were used for this study. The animals were placed randomly and allocated to treatment groups in polypropylene cages with paddy husk as bedding. Animals were housed at a temperature of 24 ± 2oC and relative humidity of 30–70%. A 12:12 light: day cycle was followed. All the animals were allowed to free access to water and fed with standard commercial pelleted chaw (M/s.Hindustan Lever Ltd., Mumbai). All the experimental procedures and protocols used in this study were reviewed by (IAEC) Institutional Animal Ethics Committee (932/a/06/CPCSEA) of Sri Lakshminarayana Institute of Medical Sciences, Pondicherry and were in accordance with the guidelines of the IAEC. Experimental protocol Experimental model of cataractogenesis was induced in rats by feeding naphthalene (Umamaheswari et al., 2011) at a dose of 0.5 g/kg orally for the first 3 days followed by 1 g/kg thereafter for a period of 25 days. Animals were divided into 5 groups consisting of six animals each. Group I received liquid paraffin (5 ml/kg b.w. orally) and served as the solvent control. Group II received naphthalene (0.5 g/kg b.w., orally for first 3 days and 1 g/kg

thereafter for 25 days) and served as the cataract control. Group III received the standard drug vitamin E at a dose of 50 mg/kg b.w., orally along with naphthalene and served as the positive control. Groups IV and V received the E. coronaria leaf extract orally at a dose of 200 and 400 mg/kg b.w. respectively simultaneously with naphthalene. All the test drugs were administered for a period of 28 days. Examination of the eyes The eyes of the rats were examined using an ophthalmoscope for morphological changes in the lens. Examination was performed after dilatation of the pupil with 1% tropicamide solution. Cataract formation was scored according to different stages. Stage 1: Clear normal lens, Stage 2: Peripheral vesicles, Stage 3: Peripheral vesicles with cortical opacities, Stage 4: Diffuse central opacities, Stage 5: Opacity involving the entire lens (Mature cataract). Cataract formation was considered complete (stage 5) when the red fundus reflex was no longer visible through any part of the lens and the lens appeared dull white to the naked eye. Percentage incidence of cataract was calculated using the following formula (Vats et al., 2004).

% Incidence = No of animals in each stage × 100 Total no. of animals

Opacity index was calculated using the following formula (Fukushi et al., 1980), Opacity Index = No. of eyes in each stage × Stage of the eye Total no. of eyes

RESULTS Anticataract activity of Ervatamia coronaria leaf extract on naphthalene induced cataractogenesis in rats was studied and the percentage incidences of cataract and opacity index were observed. Effect of E. coronaria leaf extract on % incidence of cataract on 28th day in naphthalene induced cataract rats are shown in table. 1.

Ophthalmic examinations of the normal control lenses in the eyes l were normal throughout the duration of experimental period. The animals treated with naphthalene showed varying degree of cataractogenic changes as indicated by about 72.54% in stage 4 and 45.66% in stage 5 on the 28th day of drug treatment. At the end of drug treatment, the animals treated with the extract at a dose of 200 mg/kg b.w. showed 50% of animals in stage 2,

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18.22% in stage 3 and 45.16% in stage 4 cataract, whereas the group treated with the extract at a dose of 400 mg/kg b.w. showed 50% of animals in stage 2, 25.11% in stage 3 and 18.14% in stage 4 cataract. Vitamin E the standard drug showed 18.19% of animals in

stage 1, 60.23% in stage 2 and 35.32% in stage 3. None of the animals treated with the E. coronaria leaf extract and the standard drug vitamin E showed stage 5 mature cataract at the end of the experiment.

Table 1. Effect of Ervatamia coronaria leaf extract on % incidence of cataract on 28th day in naphthalene induced cataract rats Drug Treatment Stage 1 Normal Control Liquid Paraffin (10 ml/kg) Cataract Control Naphthalene (1 g/kg) Vitamin E (50 mg/kg) Ervatamia Coronaria Extract (200 mg/kg) Ervatamia Coronaria Extract (400 mg/kg)

% Incidence of Cataract Stage 2 Stage 3 Stage 4

Stage 5

100

72.54

45.66

18.19

60.23

35.32

50.00

18.12

45.16

50.00

25.11

18.14

Table 2. Effect of Ervatamia coronaria leaf extract on opacity index in naphthalene induced cataract rats Opacity Index th th Drug Treatment 4 day 7 day 14th day 21st day 28th day Normal Control Liquid Paraffin (10 ml/kg) Cataract Control Naphthalene (1 g/kg) Vitamin E (50 mg/kg) Ervatamia Coronaria Extract (200 mg/kg) Ervatamia Coronaria Extract (400 mg/kg)

0.45

0.62

2.47

3.55

4.98

0.18

0.72

1.55

2.24

2.55

1.72

1.54

1.22

1.56

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Effect of E. coronaria leaf extract on opacity index in naphthalene induced cataract rats are shown in table 2. Treatment with naphthalene showed an increase in the opacity index from 0.45 on the 4th day, 0.62 on the 7th day, 2.47 on the 14th day, and 3.55 on the 21st day followed by complete opacification (opacity index 4.98) on the 28th day. The groups treated with the E. coronaria leaf extracts at a dose of 200 and 400 mg/kg b.w. showed a decrease in opacity index (1.72 and 1.56 respectively) when compared to naphthalene control. There was a marked reduction in opacity index (1.55) of the vitamin E treated group when compared to the naphthalene control.

rats. The cataract induced by naphthalene treatment in rats were confirmed by varying degree of cataractogenic changes and an increase in opacity index with complete opacification at the end of the 4th week. The free radical involvements in the generation of cataract have been evidenced (Kothadia et al., 2011). The in vitro antioxidant and free radical scavenging activities of E. coronaria has already been reported (Malaya Gupta et al., 2004). The anti cataract activity possessed by E. coronaria leaf extract may be due to the presence of flavonoids the well known free radical scavengers.

DISCUSSION

It can be concluded that the Ervatamia coronaria leaf extract possess anticatractogenesis activity against naphthalene induced cataract in rats. According to the results obtained from the study it may be inferred that E. coronaria reversed the cataract induced by naphthalene in rats. To the best of our knowledge this is the first report about in vivo activity of E. coronaria in a specific eye disorder and seems to raise some concern about the traditional indication of this species. A certainly further study needs to be carried out in order to prove its actual mechanism of action.

Cataract is a visual impairment that occurs due to the opacification of crystalline lens. It affects around 17 billion people worldwide, although incidence of cataracts is increasing day by day among the elderly persons. Still today except surgery no other effective treatments have been successfully developed for cataract (Piyush Patel et al., 2012). From a public health perspective, it is important to identify the risk factors that affect the development and progression of cataract. Ervatamia coronaria possess significant amount of flavonoids and a potent antioxidant has been studied against naphthalene induced cataract in

CONCLUSION

REFERENCES Bunce GE, Kinoshita J, Horwitz J (1990). Nutritional factors in cataract. Annu Rev Nutr. 10: 233–254. Da Sil Va NH, De Melo AA, Casado MMCV, Hendriques AT, Wandscheer DC, Da Sil Va OE (1984). Indole alkaloids with potential endocrine activity. Rev. Inst. Antibiot. Univ. Fed. Pernambuco. Recife. 22: 27–32.

Daniel

M, Sabnis SD (1978). Chemotaxonomical studies on Apocynaceae. Indian J Exp Biol. 16: 512–513.

Dhar ML, Dhar MM, Dhawan BN, Mehrotra BN, Ray C (1968). Screening of Indian plants for biological activity: Part - I. Ind J Exp Biol. 6: 232–247.

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Fukushi S, Merola LO, Kinoshita JH (1980). Altering the course of cataracts in diabetic rats. Invest Ophthamol Vis Sci. 19: 313–315. Haque SE, Gilani KM (2005). Effect of Ambroxol, Spirulina and Vitamin E in naphthalene induced cataract in female rats. Ind J Physiol Pharmacol. 49: 57– 64.

Pawelka KH, Stoeckigt J (1983). Indole alkaloids from cell suspension cultures of Tabernaemontana divaricata and Tabernaemontana iboga. Plant Cell Rep. 22: 105–107. Piyush

Patel, Nurudin Jivani, Shailesh Malaviya, Tushar Gohil, Yagnik Bhalodia (2012). Cataract: A major secondary diabetic complication. Int Cur Pharm J. 1: 180–185.

Henriques AT, Melo AA, Moreno PRH, Ene LL, Henriques JAP, Schapoval EES (1996). Ervatamia coronaria: Chemical constituents and some pharmacological activities. J Ethnopharmacol. 50: 19– 25.

Sharma P, Cordell GA (1988). Heyneanine hydroxyindolenine, A new indole alkaloid from Ervatamia coronaria var. plena. J Nat Prod. 51: 528–531.

Hsu YT (1967). Study on the Chinese drugs used as cancer remedy. J Southeast Asian Res. 3: 63–66.

Taesotikul T, Panthong A, Kanjanapothi D, Verpoorte R, Scheffer JJC (1989). Hippocratic screening of ethanolic extracts from two Tabernaemontana species. J Ethnopharmacol. 27: 99–106.

Kirtikar KR, Basu BD (1975). Indian Medicinal Plants, V 2, Bishen Mahendra Pal Singh Dehradun, India. 842–844. Kothadia AD, shenoy AM, Shabaraya AR, Rajan MS, Viradia UM, Patel NH, (2011). Evaluation of cataract prevention action of phycocyanin. Int J Pharm Sci and Drug Res. 3: 42–44. Malaya Gupta, Mazumdar UK, Gomathi P, Sambath Kumar R (2004). Antioxidant and Free Radical Scavenging Activities of Ervatamia coronaria Stapf. Leaves. Iran J Pharm Res. 2: 119–126.

Umamaheswari M, Asokkumar K, Lalitha V, Sivashanmugam AT, Subhadradevi V (2011). Anticataract and antioxidant activities of Citrus aurantium L. peel extract against naphthalene induced cataractogenesis in rats. J Phar Res. 4: 680–682. Vats V, Yadav SP, Biswas NR, Grover JK (2004). Anticataract activity of Pterocarpus marsupium bark and Trigonella foenum-graecum seeds extract in alloxan diabetic rats. J Ethnopharmacol. 93: 289–294. Yamamoto T, Takahashi H, Sakai K, Kowithayakorn T, Koyano T(1997). Screening of Thai plants for anti-HIV-1 activity. Natural Med. 51: 541–546.

Source of Support: Nil

Conflict of Interest: None Declared

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Research article STUDIES ON TRADITIONAL HERBAL PEDIATRICS PRACTICES IN JAISINGHPUR, DISTRICT KANGRA (HIMACHAL PRADESH, INDIA) Rawat Dhiraj S1, Kharwal Anjna D2 Department of Botany, R.K.M.V. – Shimla, Himachal Pradesh, India Department of Botany, Govt. Degree College- Dharamshala. Himachal Pradesh, India *Corresponding Author: Email: rawatdhirajhpu@gmail.com 1 2

Received: 26/02/2013; Revised: 01/04/2013; Accepted: 02/04/2013

ABSTRACT Plants are the basis of life on earth and are central to people‟s livelihood. Glimpses of our knowledge in ethnomedicine are available in vedic texts and there is an inextricable link between indigenous culture and biodiversity as areas of high biodiversity are often found on indigenous community‟s lands. The local communities and rural populace of Jaisinghpur is highly dependent on nature for meeting their healthcare needs and has a repository of accumulated experience and knowledge of prevailing vegetation of the region. Medical ethno–botany forms a major part of medicinal aspects of aboriginal child care. 70 % of world population uses herbal traditional remedies in treatment of sick and injured children. Indigenous herbal practices related to child–care provide invaluable knowledge and aid in making best use of natural resources as it is dynamic in dissemination and scientific in indigenous experimentation. Present study includes 21 plants (15 dicots and 6 monocots) belonging to 16 families used as herbal remedies for child–care, while 2 plant spp. are used along with other plant resources in herbal preparations. Among various plants, fruits of 7 species (32%), seeds of 6 species (29%), leaves of 5 species (24%), peduncle, bark and rhizome of one plant (5%) each is used predominantly for child–care by the rural populace of the study area. Mostly, the people of age groups 41–60 years (AG–3) and >60 (AG–4) years mostly ladies, are aware of these herbal practices. Local communities not only use these plants but also care for their conservation and protection; thus contributing towards sustainable development. KEY–WORDS: Ethnobotany, Biodiversity, Child–care, Herbal remedies, ODA (Observed density & availability), Phenological pattern, TIV (Total importance value), Pediatrics. ABBREVIATIONS: (AG– Age group/s)

Cite this article: Rawat Dhiraj S, Kharwal Anjna D (2013), STUDIES ON TRADITIONAL HERBAL PEDIATRICS PRACTICES IN JAISINGHPUR, DISTRICT KANGRA (HIMACHAL PRADESH, INDIA), Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 219–230

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 219–230

INTRODUCTION The term “Ethno–botany” was coined by J.W. Harshberger in 1895, meaning “the study of plants used by primitive and aboriginal people” (Anonymous, 1895). Since then, the subject has been variously defined and interpreted by different workers as its discipline began to follow multidisciplinary approach combining a diversity of knowledge bases and methods through the use of anthropological methods (Robbins et al., 1916; Schultes & Reis, 1995). Plants are the basis of life on earth and are central to people‟s livelihood. Glimpses of our knowledge in ethno–medicine are available in vedic texts (Jain, 1987). Undeniably, there is an inextricable link between indigenous culture and biodiversity as areas of high biodiversity are often found on indigenous community‟s lands and in their water bodies (Alcorn, 1996). The 15th session of the General Assembly of IUCN held in Christchurch, New Zealand, in October 1991, recognized the importance of the cultural heritage of mankind and the role of traditional cultures in conservation of nature (McNeely & Pit, 1985). Agenda 21 of the Rio Earth Summit (1992) stated that indigenous people have a vital role in environmental management and development because of their knowledge and traditional practices. Ethno–botanical information in the form of folklore is passed through generations in certain restricted and remote habitations (Chauhan, 1999; Choudhary et al., 2008; Ganesan, 2008; Saini, 1996). “Jaisinghpur” (592m), one of the tehsils of district Kangra in Himachal Pradesh has common boundaries with districts Mandi and Hamirpur (Fig. 1). The word “Jaisinghpur” is derived from the name of a famous king Raja Jai Singh who is believed to be a great warrior of “Rajgir” dynasty. Still the name of the legislative constituency is “Rajgir” (reserved for S.C.) which comprises most of the areas of tehsil “Jaisinghpur”. The town “Jaisighpur” is located on the bank of river “beas” with a population of 1,273 while the population of tehsil is 58,623. Tehsil “Jaisinghpur” is full of natural water resources and is a combination of

greenery and water, thus, has given the tehsil a distinctive look, located at an altitude between 500–1800 m above MSL between 31°53′55′′N/76°35′58′′E latitudes. The area is a combination of the plains and the hills and blessed with remarkable natural beauty and high ranges of Dhauladhar mountains at the backdrop with tops remain snow covered for most part of the year. The natives are the Kangri people and the local language is „Kangri‟. The majority of the people are „Hindu‟. Traditional dresses of men are „kurta‟, „pyjamas‟ with a woolen jacket in winter. Women generally wear „salwaar‟, „kameez‟ along with „chunni‟ („chaddru‟ in local language). Maize, wheat and paddy are the main staple foods of the rural populace and the villagers are very fond of butter, milk, curry preparations and pickles. The place unfolds four broad seasons with winters spreading generally from December– February, summers from March–June, rainy season extending from July–September with landscape becoming lush green, and autumns from October–November. Agriculture is the main stay of the inhabitants of the area. Soil varies from sandy loams to clay. The agro– climatic conditions favour the growth of food crops such as wheat, paddy, maize, potatoes, etc. Agricultural operations are carried out in two spells. Spring crops popularly called as Rabi („Harri‟) comprise of wheat, barley, gram and oil seeds (linseed) whereas autumn crops (Kharif / „Savani‟) are maize, paddy, pulses, spices and potatoes (Balokhra, 2002). The area on the bank of river “beas” is highly fertile and is famous for vegetable cultivation. This area is known as “shukdi ka bag”. The region is also famous for its “holi” and “dussehra” festivals. “Chaugan” of “Jaisinghpur”, “Harsi”, “Laddi”, “Lambagaon”, “Ashapuri” temple, “Nagban” and “Sai” are the places of interest. The local communities and rural populace of Jaisinghpur is highly dependent on nature for meeting their healthcare needs. The rural populace of the region has a repository of accumulated experience and knowledge of prevailing vegetation of the region. They have

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a deep belief on their nature folklore medicines for remedies and they rely exclusively on their herbal cure. Recently, considerable attention has been paid to utilize eco–friendly plant based products for the prevention and care of different human diseases. In India over 6,000 plants are in use in traditional folk and herbal medicine, which constitutes about 75% of the medical needs of third world countries (Rajshekharan, 2002). Similarly, herbal

medicines for infants and child–care are not exceptions in the study area. The women folks of the region play a vital role in use and mobilization of biodiversity based knowledge system. Medical ethnobotany forms a major part of medicinal aspects of aboriginal child care. 70 % of world population uses herbal traditional remedies in treatment of sick and injured children (Pearn, 2005).

Fig 1. Map of tehsil Jaisinghpur (district Kangra, H.P.) showing main locations visited for survey.

SIGNS 1- Lambagaon block -Locations

There is an inextricable nexus between aboriginal men, women and land in which they live. Aboriginal women in traditional communities use a sophisticated botanical material media in the treatments of sick and injured children. Drugs and medicaments used in treatments are obtained from various plant parts usually as fresh preparations in the form of infusions, macerations, decoctions etc. and are rarely stored (Ganesan, 2008). The multipurpose and broad–spectrum use of plants to treat symptoms and symptom complexes in context of preventive medicine for child care is the heart of discussion. Internationally, one of the best works is that of Pearn, 2005, which throws a light on traditional pediatric practices in Australia and the work of Allen & Hatfield,

Source www.google.co.in

2004 which emphasized on ethnic studies of Britain and Ireland and that of Salah & Nyunda, 2012 which emphasized on pre-natal care. The information on floristic and ethno– botanical studies related to child–care in India is scattered meager (Babu, 1998; Borthakur, 1993; Choudhary et al., 2008; Ganesan, 2008; Goyal et al., 2011; Joshi, 1989; Pal et al., 2000; Qureshi, 2007; Rajshekharan, 2002; Robbins et al., 1916; Saini, 1996; Sen et al., 2008). Ethnobotanical information on child care in Himachal Pradesh (H.P.) and district Kangra is meagre in literature (Chauhan, 1999; Dhiman, 1976). Inspite of rich floristic diversity and cultural values, nothing is available in literature on the floristic and ethnobotanical information of the region (study area). Keeping this in

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mind, the present study had been undertaken with the following objectives – (a) to collect and identify the plant spp. used for child–care along with their flowering and fruiting seasons (b) to study the phonological pattern of collected plants (c) to calculate total importance value (TIV) and observed density availability (ODA) of plant spp. (d) to know about the effective age groups involved in herbal practices. Since time immemorial, the Himalaya has influenced the life and culture of the diverse ethnic communities living all along the length of its mountainous chain. Keeping this in mind it is pertinent to document this knowledge for future generations. METHODOLOGY Intensive ethnobotanical exploration were undertaken in the rural pockets of tehsil Jaisinghpur, district Kangra (H.P.). The villages selected for study are: Draman (900 m), Dhupkyara (720 m), Laddi (1290 m), Nee (910 m), Langa (845 m), Arth (835 m), Bhedu (790 m), Bhaati (750 m), Jalag (840 m), Nahlana (785 m), Saped (810 m), Nakki (930 m), Tamru (920 m), Bhodi (890 m), Hadot (915 m), Kosri (760 m), Ropari (750 m), Suan (750 m), Tarapad (925 m), Tikri (620 m), Kamanda (630 m), Dwata (980 m), Sai (800 m), Harsi (550 m), Kathla (590 m) and Bardama (690 m). The field tours were planned in such a way so as to collect the ethnobotanically interesting species used for infants and child–care either in flowering or fruiting stage. Herbarium of collected plants was prepared following Jain & Rao, 1978. For a better understanding of local beliefs, habits and uses of plants, different categories of people like family heads, healers, old experienced and knowledgeable informants, especially old ladies were repeatedly interviewed. Specific questions based upon Proforma designed by Jain & Rao, 1978 were asked and the resultant information was recorded in the ethno-botanical field notebook along with the name of locality and local name. Botanical identification of the selected species

was first done with the help of regional floras (Chauhan, 1999; Chowdhery & Wadhwa, 1984; Dhiman, 1976; Hooker, 1897). For more information three basic approaches were adopted following Phondani et al., 2010:  An interview based approach– Questions from informants on infant and child–care plants mainly from old experienced people, especially old ladies as they were more aware of the child–care plants.  An inventory based approach– An inventory based approach is followed on following questions:  Whether whole plant or plant parts are used?  Which age group is more aware to these herbal remedies?  The season of flowering and fruiting  TIV (Total importance value) of these plants  The density of plants in the region  Whether the plant is used for one disease or for more than one disease?  An interactive discussions approach with communities–  How to use plants?  Are they better than market products or not?  Are they used singly or in combination?  Are all the plants or plant parts available in nature or some of them are taken from market? While collecting the plant specimens, their uses related to child–care and their local names were also ascertained and recorded carefully in the field notebook with the assistance of local informants. The data were verified in different regions among the interviewers and showing the same plant sample, and even with the same informants on different occasions. Ethno– botanical lore was considered valid if at least three informants made similar comments. Four age groups (AG) were investigated i.e. AG–1 (0–20 years), AG–2 (21–40 years), AG–3 (41–

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60years) and AG–4 (>60 years) to find out the impact of ethno–botanical lore. Men and women of local communities are interviewed separately to find out the gender based herbal knowledge. Phenological pattern of the plants were observed to find out the seasonal variation while ODA (Observed density availability) was observed according to Sood et al., 2012 in which plants were classified into abundant, considerable and rare extent. Nomenclature of these taxa were confirmed from Bennet, 1986 and Wielgorskaya,1995. Economic valuation of all the presently recorded ethno–botanical species was also carried out to calculate the total importance values (TIV) on the sum basis of parameters like life cycle strategy, periodicity of use, habit, availability throughout year and uses as per detailed methodology outlined by Belal & Springuel, 1996. OBSERVATIONS The local communities of tehsil Jaisinghpur of district Kangra (H.P.) use 21 plant spp. in 18 different types of herbal practices related to child–care. These local communities are a rich repository of traditional knowledge, so a sincere effort has been made to get the information on these herbal practices which are: Bark of Ficus religiosa is cut into pieces and to this seeds of “puthkanda” Achyranthes bidentata are added along with peduncles of “challi” maize (Zea mays). The mixture is burned to ash. Ash is mixed with borax (Suhaga) and honey. Half teaspoon is recommended thrice a day (one week) for bronchitis, cough and congestion. (AG–3, 4). 1–2 small holes are created in unripe fruits of “rada” (Randia dumetorum) with the help of thorns of the same and these holes are filled gently with the milk (latex) of Ficus palmata. These fruits are sealed with kneaded “kanak” wheat flour and finally the sealed fruits are roasted in a fire–place “chullah”. Powder of

roasted fruits along with honey (½ teaspoon twice a day for a week) is highly recommended against bronchitis, cough and congestion. (AG– 3, 4). Leaves of “Kouru” Roylea cinerea are crushed with the help of a clean pastel and mortar. Juice is filtered with a fine clean cloth piece and 1–2 drops of it are used twice as nasal drops for three consecutive days. It is considered good for cough. (AG–3, 4). A longitudinal fine cut is made into “chhuara” (Fruit of Phoenix dactylifera) and the seed is taken out. The left out cavity so created, is filled up with powdered seeds of “chhoti ellaichi” (Elettaria cardamomum) and the fruit is sealed with thread. It is roasted in a fire–place “chullah”. ½ of the fruit is taken with milk at bed time for 1–2 months for checking frequent urination in infants and children. (AG–3, 4). Rhizomes of “barain” Acorus calamus are washed with water and its paste along with honey is taken thrice a day for a week against cold, cough and bronchitis. (AG–2, 3). 4–5 small stones having smooth surface are thoroughly washed and are dumped at fire– place “Chullah” for 20–30 min. About 100 ml of drinking water is taken in a clean bowl and 1 tablespoon full of “ajwain” seeds (Trachyspermum ammi) is added to it. Hot stones are taken out from fireplace with the help of “chimtah”– a tong like household article and ash is removed with the help of cotton cloth. The stones are dipped in water taken in the bowl. The fluid in the bowl is filtered and the filtrate so obtained is locally known as “gitt– juanhe” as the word “gitt” is used for stones and “juanhe” for ajwain (Trachyspermum ammi) in local „Kangri‟ language. 2–3 teaspoons of “gitt–juanhe” is prescribed thrice a day for 3–4 days to check stomach ailments in infants. (AG–4). Fruits of “jaiphal” (Myristica fragrans) are rubbed on a clean stone with water and the paste so obtained is prescribed for children to check cough and congestion thrice a day for 4

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days and the same is also applied over nose for the same. (AG–2, 3). Decoction of “ajwain” seeds (Trachyspermum ammi) is highly prescribed for infants and children for stomachache. Usually 5–10 tablespoons are given to infants and children after every 2–3 hours until relief. (AG–1, 2, 3, 4). Decoction of aerial parts (leaves, stem, inflorescence and even seeds) of “tulsi” (Ocimum sanctum) is highly prescribed for couch and fever. (AG–2, 3, 4). The fruits of “harad” (Terminalia chebula) are rubbed over a clean stone or in “kundi"– a mortar type household article and a pinch of its paste along with lukewarm water is prescribed thrice a day against constipation until relief. (AG–2, 3, 4). Decoction of “kadwi–saunf ” seeds (Foeniculum vulgare) is prescribed for infants and children for stomachache. Usually 1–2 tablespoons are given to infants. (AG–1, 2, 3, 4). Decoction of “chhoti ellaichi” (Elettaria cardamomum) along with sugar is considered good for lung ailments. (AG–3, 4). Fruit poultice of “dodey” (Sapindus mukorosii) is made and a thick layer of it is applied over a clean cotton cloth and this cloth is tied around mumps and considered one of the best traditional remedy against it. (AG–3, 4). Seeds of “til” (Sesamum orientale) along with jaggery and “soya” (Anethum graveolens L.) are used to make traditional “laddu” which are given to infants to check bed wetting. Similarly ripe dried fruits of “Chhuara” (Phoenix dactylifera) are prescribed for the same. (AG–3, 4). Onion “pyaz” (Allium cepa) juice is mixed with mustard “sarson” (Brassica campestris L.) oil. 1–2 drops thrice a day (for a week) is recommended for ear itching. (AG–2, 3, 4).

Patients exposed to the smoke on burning of “naule ra lingna” (aerial parts of Verbascum thapsus) are considered to have relief from measles. (AG–4). Pieces of leaves of “kwarya” (Aloe vera) are heated gently. Each piece is cut gently into two pieces to expose the gel of leaves. Each piece singly or in combination with “haldi” i.e. turmeric powder (Curcuma angustifolia Roxb.) is used against muscle pull. (AG– 1, 2, 3, 4). “Jaiphal” Myristica fragrans” is rubbed over a soft clean stone and the paste of it is applied over cotton plugs. These cotton plugs are placed in mustard (“sarson”– Brassica campestris) oil taken in an earthen pot “diya” and enlightened with fire. The pot is placed in the corner of a dark room and a brass plate is placed on it for 3–4 hours. The deposited carbon on brass plate is collected. It is mixed with cow‟s ghee and small quantity of “jaiphal” paste. The obtained product is locally called as “kajjal” which is applied in eyes to check any infection. It is also used for healthy and beautiful eyes. (AG– 3, 4). RESULTS & DISCUSSION: The study of ethno–medical systems and herbal medicines as therapeutic agents of a paramount importance in addressing health problems of traditional communities and third world countries as well as industrialized societies (Rajshekharan, 2002; Saini, 1996). The present study yielded interesting data which provides information of the 21 plants used for child–care in tehsil Jaisinghpur of district Kangra (Himachal Pradesh). The plants are used in 18 different herbal practices of the region in which these plants are used either singly or in combination. These plants are arranged in alphabetical order; with their local name, flowering and fruiting seasons, ODA (Observed density & availability) and part / parts used as in table 1. Present study includes 21 plants (16 dicots and 5 monocots) (Table 1), used as herbal remedies for child–care belonging to 16 families. The predominant families are

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Liliaceae with 3 spp, Moraceae, Apiaceae and Poaceae with 2 spp each. Ficus with 2 spp is the dominant genus. Among various plants, fruits of 7spp (32%), seeds of 6 spp (29%), leaves of 5 spp (24%), peduncle, bark and rhizome of one plant (5%) each is used predominantly for childâ&#x20AC;&#x201C;care by the rural populace of the study area (Figure 2). 12 plant spp are used singly while 5 plant spp. are used in combination. One plant i.e. Ellettaria cardamomum is used along with sugar for lung ailments.

ODA (Observed Density Availability) reveals that 8 plant spp are in abundant extent while 9 spp. are in considerable extent. One cultivated plant (Gossypium arboreum) is in rare extent and the cotton (seed surface hairs) of this can be purchased from the market so that the plant resource can be used in a sustainable manner in the study area. Phenological pattern of plants suggest that most of the plants are in flowering and fruiting stage during rainy, summer and spring seasons (Fig. 3).

Fig.2. Various plant parts used in herbal remedies in tehsil Jaisinghpur of district Kangra (H.P.). Plant parts in herbal rem edies

Fruit s

32%

Seeds 24%

Leaves 29%

Bark Rhizome Peduncle

Fig. 3. Phenological pattern of ethnobotanical plants in tehsil Jaisinghpur of district Kangra (H.P.). Phenological pattern

No. of plants

12 10 8 6 4 2 0 Winter

Spring

Summer

Rainy

Seasons

Table 1

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 219–230 Table -1 List Of Plants Used as Herbal Remedies for Child–Care in Tehsil Jaisinghpur of District Kangra (Himachal Pradesh) S.N.

Botanical Names and Family

Local Names

Flowering Seasons

Achyranthes bidentata Blume (Amaranthaceae)

Puthkanda

Acorus calamus L. (Liliaceae)

Fruiting

ODA

Part/s Used

Mar.–Oct.

+++

Seeds

Barain

June–July

Rhizome

Allium cepa L. (Liliaceae)

Pyaz

Dec.–Mar.

Scales

Aloe vera (L.) Webb. & Berthel. (Liliaceae)

Kwarya

Throughout year

Leaves

Brassica campestris L. (Brassicaceae)

Sarson

Oct.–Mar.

+++

Seeds

Foeniculum vulgare Mill. (Apiaceae)

Kadwi Saunf

Oct.–Mar.

Seeds

Ficus palmata Forsk. (Moraceae)

Dhuda, Phegda

April–Sept.

+++

Latex

Ficus religiosa L. (Moraceae)

Peepal

Mar.–Oct.

+++

Bark

Gossypium arboreum L. (Malvaceae)

Kapaa

Jul.–Sept.

Seed–hairs

10.

Ocimum sanctum L. (Lamiaceae)

Tulsi

Aug.–Sept.

+++

Aerial Parts

11.

Randia dumetorum (Retz.) Lam. (Rubiaceae)

Rada

April–Sept.

Fruits

Roylea cinerea (D. Don) Baill. (Verbenaceae)

Kouru

Mar.–Nov.

Leaves

Sapindus mukorosii Gaertn. (Sapindaceae)

Dodan, Reetha

April–Sept.

+++

Fruits

14.

Sesamum orientale L. (Pedaliaceae)

Til

July–Sept.

Seeds

15.

Terminalia chebula Retz. (Combretaceae)

Harad

April–June

+++

Fruits

16.

Trachyspermum ammi Sprague (Apiaceae)

Ajwain

Oct.–Mar.

Seeds

17.

Verbascum thapsus L. (Scrophularaceae)

Naule leengna

Jan.–April

Aerial parts

18.

Zea mays L. (Poaceae)

Challi

June–Sept.

+++

Peduncle

19.

Phoenix dactyllifera L. (Arecaceae)

Khajur

From market

Fruits

20.

Myristica fragrans Houtt. (Myristicaceae)

Jaiphal

From market

Fruits

21.

Elettaria cardamomum (Zingiberaceae)

Chhoti–ellaichi

From market

Fruits

12.

13.

(L)

Maton

ODA – Observed Density Availability + + + – Abundant + + – Considerable extent + – Rare extent Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 219â&#x20AC;&#x201C;230 Table 2 - Total importance value (TIV) of ethnobotanical plants in tehsil Jaisinghpur of District Kangra (Himachal Pradesh)

S.N. Botanical Names and Family

Life Periodicity Habit Availability cycle of use throughout strategy year

Uses TIV (%)

Achyranthes bidentata Blume (Amaranthaceae)

Acorus calamus L. (Liliaceae)

Allium cepa L. (Liliaceae)

Aloe vera (L.) Webb. & Berthel. 2 (Liliaceae)

Brassica campestris L. (Brassicaceae)

Foeniculum vulgare Mill. (Apiaceae)

Ficus palmata Forsk. (Moraceae)

Ficus religiosa L. (Moraceae)

Gossypium arboreum L. (Malvaceae)

10.

Ocimum sanctum L. (Lamiaceae)

11.

Randia dumetorum (Retz.) Lam. (Rubiaceae)

12.

Roylea cinerea (D. Don) Baill. (Verbenaceae)

13.

Sapindus mukorosii Gaertn. (Sapindaceae)

14.

Sesamum orientale L. (Pedaliaceae)

15.

Terminalia (Combretaceae)

chebula

Retz. 4

16.

Trachyspermum (Apiaceae)

ammi

Sprague 2

17.

Verbascum thapsus L. (Scrophularaceae)

18.

Zea mays L. (Poaceae)

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Fig. 4. Herbal practices in different age groups in Tehsil Jaisinghpur. Herbal practices in different age groups 18 16 14 12 No. of herbal 10 practices 8 6 4 2 0 AG-1

AG-2

AG-3

AG-4

>60 only

Age groups

Out of discussed 18 herbal practices, 3 herbal practices were known to the population of 0–20 years i.e. age group1 (AG–1); while 9 to age group 2 of 21–40 years (AG–2). Mostly, the people of age groups 41–60 years (AG–3) and >60 years were aware to 17 practices while 2 practices are restricted to age group 4 (AG–4) i.e. > 60 years (Fig. 4). From this it is crystal clear that these practices are mostly restricted to AG–3 and AG–4 while few of them are restricted to old ladies i.e. this knowledge is fast depleting in younger generations so it is pertinent to document this invaluable eco– friendly herbal remedies. Gender wise analysis reveals that 14 practices (5–18) are known to both men and women of the study area while first 4 practices (1–4) are restricted only to women of different communities of the region (Fig.4). Two plant spp. i.e. Curcuma angustifolia roxb. (powdered rhizome) and Anethum graveolens L. (seeds) are used with other plant resources in some of the herbal practices. These plants are also cultivated in the study area and can be purchased from the market. Due to the utmost importance of these plants in the region, local communities not only use these plants but

also care for their conservation and protection; thus contributing towards sustainable development. Statistically, the total importance value (TIV) reveals that Terminalia chebula tops the list with TIV of 85%. Ficus reliogosa, Ficus palmata have 80% TIV while Roylea cinearea and Sapindus mukorosii have TIV of 75%. Gossypium arboretum and Zea mays have lowest TIV of 40% with respect to medicinal values for child–care. (Table II) CONCLUSION Indigenous herbal practices related to child–care provide invaluable knowledge and aid in making best use of natural resources as it is dynamic in dissemination and scientific in indigenous experimentation. In the modern days of technological advancement, this knowledge is falling prey to the lure of modernization and urbanization. Negligible efforts have been undertaken to understand the scientific basis of the knowledge. It is recommended that the documentation of indigenous herbal practices should be included in the curricula of environment and sustainable development as a cross cutting issue.

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REFERENCES Alcorn, JB (1996). Is biodiversity conserved by indigenous people? In: Ethnobiology in HumanWelfare. SK Jain (Ed.), New Delhi: Deep Publ., pp 233–238. Allen, DE & Hatfield, G (2004). Medicinal plants in folk tradition : An ethnobotany of Britain & Ireland. UK: Timber Press. Anonymous (1895). Some new ideas, The plants cultivated by aboriginal people and how used in primitive commerce. Philadelphia: The Daily Evening Telegraph, vol. 64, no 134. Babu R (1998). Mother & Child Care. J. Amruth 2(6) : 98–102. Balokhra, JM (2002). The Wonderland Himachal Pradesh. New Dehli: H.G. Publication. Belal, Ahmed E & Springuel I (1996). Economic value of plant diversity in arid environments. Nature & Resources 32 (1): 33–39. Bennet, SSR (1986). Name Changes in Flowering Plants of India and Adjacent Regions. Dehradun, India: Triseas Publ. Borthakur, SK (1993). Native phytotherapy for child and women diseases from Assam. Ethnobotany 5 (87): 87–91. Chauhan, NS(1999). Medicinal and Aromatic Plants of Himachal Pradesh. New Delhi: Indus Publ. Co. Choudhary, K, Singh, M & Pillai, U (2008). Ethnobotanical survey of Rajasthan – An uptade. Am Eurasian J. Bot. 1(2): 38–45. Chowdhery, H J & Wadhwa , BM (1984). Flora of Himachal Pradesh. Calcutta: Vol. 1–3. Bot. Surv. India.

Dhiman, DR (1976). Himachal Pradesh Ki Vanoshdhiya Sampada. Dharamsala, H.P.: Imperial Printing Press. Ganesan, S (2008). Traditional oral care medicinal plants survey of Tamil Nadu, Natural Product Radiance 7: 166–172. Goyal, M, Sasmal, D &Nagori, BP (2011). Review on medicinal plants used by local community of Jodhpur district of Thar desert. Int. J. Pharmacology 7: 333–339. Hooker, JD (1872–1897). The Flora of British India. Allahabad: Vol. I–VIII, Lalit Mohan Basu. Jain, SK & Rao, RR (1977). A Handbook of Field and Harbarium Methods. New Delhi: Today‟s & Tomorrow‟s Printers & Publ. Jain, SK (1987). A Manual of Ethnobotany. Jodhpur : Sci. Publ. Joshi, P (1989). Herbal drugs in Rajasthan– from childbirth to child–care, Ethnobot. 1:77–87. McNeely, JA & Pit, D (Eds.) (1985). Culture and Conservation, The Human Dimension in Environmental Planning. London, New York, Sydney: Croom Helm. Pal, DC, Guha, A & Sen, R (2000). Medicinal Plants and Plant Products Used in Children Diseases among Aboriginal in India. In: Ethnobotany and Medicinal Plants of Indian Subcontinent. JK Maheshwari (Ed.), Jodhpur: Scientific Publishers. Pearn, J (2005). The world‟s largest surviving pediatric practices: some themes of aboriginal medical ethnobotany in Australia. J. Paediatr. Child Health 41(5–6): 284–290.

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Phondani, PC, Maiknuri, RK, Rawat, LS, Farooquee, NA, Kala, CP, Vishwakarma, SCR, Rao, KS & Saxena, KG (2010). Ethnobotanical uses of plants among the Bhotiyal tribal communities of Niti valley in Central Himalaya, India. Ethnobot. Res. & Application 18 : 233–244. Qureshi, RA, Ghufran, MA, Gilani, SY, Sultane, K & Ashraf. M (2007). Ethnobotanical survey of selected medicinal plants of Sudhan gali and Ganga chotti hills, district Bagh, Kashmir. Pak. J Bot. 39 (1) : 2275– 2283.

Saini VK (1996). Plants in the Welfare of Tribal Women and Children in Certain Areas of Central India. In: Ethnobiology in Human Welfare. SK Jain (Ed), New Delhi: Deep. Publ., pp 140–144. Salah, AM & Nyunda, LA (2012). Effect of Ruellia praetermissa extracts on erythropoiesis in pregnant women. GJMRI. 1(8): 309–314. Schultes, RE & Reis, SV (1995). Ethnobotany – Evolution of a Discipline. London:Chapman.

Rajshekharan, PE (2002). Herbal medicine, in world of science. Employment News (21–27), 3.

Sen, S, Chakraborty, R, Biplab D & Devanna , N (2008). An ethnobotanical survey of medicinal plants used by ethnic people in west and south district of Tripura, India, J. For. Res. 22 (3) : 417–426.

Robbins, WW, Herrin, JP & Freire –Marreco, B (1916). Ethnobotany of the Tewa Indians, Bureau of American Ethnology. Washington, D.C. : Bulletin, no 55. Smithsonian Institutions.

Sood, S K, Rawat, D, Rawat, S & Kumar, S (2012). A Handbook of Wild Edible Plants. Jaipur: Pointer publishers.

Source of Support: Nil

Wielgorskaya, T (1995). Dictionary of Generic Names of Seed Plants. Dehradun: B.S.M.P.S.

Conflict of Interest: None Declared

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Research article LIVER ENZYMES AND ITS ASSOCIATION WITH AGE AND SEX IN SICKLE CELL ANAEMIA PATIENTS AND HAEMOGLOBIN S TRAIT CARRIERS. Chuku L C1, Chinaka N C2 1, 2

Department of Biochemistry, University of Port Harcourt, P.M.B. 5323, Choba, Port Harcourt, Rivers State, Nigeria *Corresponding Author: cn_chinaka@yahoo.com

Received: 28/02/2013; Revised: 04/04/2013; Accepted: 05/04/2013

ABSTRACT The activities of the liver enzymes, alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase were compared between males and females of various age groups (0–25+) during sickle cell crisis and in steady state. Results show that enzyme activities increased during painful crisis. The difference in activities of the enzymes in normal homozygous HbAA and heterozygous (HbAS) blood was not statistically significant (p ≥ 0.05). The activities of the enzyme increased with age in all the genotypes studied. There was no significant difference (p ≥ 0.05) in the activities of ALT, AST and ALP between males and females for all the age groups studied. KEY WORDS: Alanine transaminase, alkaline phospatase, anaemia, aspartate transaminase, genotype, heterozygous, homozygous, sickle cell.

Cite this article: Chuku L C, Chinaka N C (2013), LIVER ENZYMES AND ITS ASSOCIATION WITH AGE AND SEX IN SICKLE CELL ANAEMIA PATIENTS AND HAEMOGLOBIN S TRAIT CARRIERS., Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 231–237

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INTRODUCTION

Electrophoresis

Sickle cell anaemia is a form of sickle cell disease in which both abnormal genes are for the formation of HbS (Konotey-Ahulu, 1974). Sickle cell haemoglobin (HbS) is the major abnormal haemoglobin. Its solubility is sufficiently altered to produce a serious disease when present in the homozygotes (White, et al., 1978). Sickle cell anaemia (SCA) has been a great clinical problem in the history of West Africans since the 7th century. The disease was first described by a Chicago physician, J.B. Herrick (Herrick, 1910). Herrick observed peculiar elongated and sickled-shaped red cells in a case of severe anaemia of an ailing West Indian student.

In all cases, blood samples from different individuals were genotyped. Different molecular species of Hb were separated from each other by electrophoresis at pH 8.4 on cellulose acetate. The separated haemoglobin bands were then stained by Ponceau S dye and identified by comparison with known haemoglobin standards separated stained in the same manner.

The term “sickle cell trait” is used to describe a person who has inherited one normal haemoglobin gene (A) from one parent and one abnormal gene (S) from the other parent. Sickle cell disease arose as a result of mutation which caused a single amino acid substitution (valine for glutamic acid) at the sixth position of the βglobin chain of the haemoglobin molecule (Acquaye, et al., 1981). This change has an adverse effect on the haemoglobin molecule. Hence, the research was undertaken to study any variation that might occur in the enzymes in normal and sickle cell subjects over a wide age range and sexes (male and female).

Assay of enzyme activity

MATERIALS AND METHODS Materials, test kits and equipments used were of laboratory standards. Blood from normal and heterozygous individuals was obtained at the University of Port Harcourt Teaching Hospital and LABMEDICA laboratory, Port Harcourt. Samples were then grouped according to their ages and sex. The normal subjects (HbAA and HbAS) were aged between 0–70 years and of both sexes. It was not possible to obtain HbSS blood from patients beyond 20 years of age. Blood was also collected from HbSS patients who were in haemolytic crisis.

Different types of Hb were separated from one another by electrophoresis on cellulose acetate paper. Separation was clear at the buffer pH of 8.4. Sixty four (64) blood samples were separated into 24 AA, 16 AS and 24 SS.

Alanine transaminase (ALT) activity: By Ratman and Frankel, (1957). Alanine transaminase activity was estimated by the procedure described in the RANDOX kit for the determination of alanine transaminase activity in serum or plasma at 546 nm. The optical density (O.D) of the reaction mixture was taken colorimetrically at 546 nm as a measure of the enzyme activity. Aspartate transaminase (AST) activity: By Ratman and Frankel, (1957). Aspartate transaminase catalyses the reaction (transamination) between αketoglutarate and L-aspartate. The oxaloacetate formed in the reaction reacts with 2, 4dinitrophenylhydrazine (DNPH), which in alkaline medium gives a red-brown colour. This is measured in a colorimeter at 546 nm. This measures the activity of aspartate transaminase. The enzyme activity was measured by a procedure described in the RANDOX GOT kit. Alkaline phosphatase (ALP) activity: By Ratman and Frankel, (1957). Alkaline phosphatase (ALP) hydrolyses pnitrophenyl phosphate in alkaline conditions to

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yield phosphate and p-nitrophenol. The nitrophenol is yellow in colour and absorbs maximally at 405 nm. The intensity of the colour due to p-nitrophenol during a fixed time is measured at 405 nm colometrically and is proportional to the ALP activity in the sample. This was estimated by the procedure described in the “BESSEY LOWRY” colour method kit for the determination of ALP activity in plasma or serum at 420 nm. RESULTS Enzyme Activity The effect of age on ALT, AST and ALP activities of HbAA, HbAS and HbSS subjects are shown in tables 3.1 as well as their steady and crisis state as shown in table 3.2.1, 3.2.2 and 3.2.3 respectively. The effect of sex on ALT, AST and ALP activities in HbAA and HbAS subjects are is illustrated in table 3.3.1 respectively. The effect of sex on AST and ALP activities of HbSS and its crisis state is shown on table 3.3.2 and 3.3.3 respectively.

DISCUSSION Alanine transaminase (ALT): The activity of ALT of sickle cell subjects was found to be higher than normal. There was however no significant difference (p ≥ 0.05) in the specific activity of ALT for normal (AA and AS) subjects. It was also observed that the activity of ALT did not increase steadily with age and did not also depend on sex. Serum ALT is highly variable in sickle cell disease. Values obtained for subjects between 11–25 years were far lower than those obtained for subjects between 6–10 years although higher than the values obtained for subjects between 0–5 years. This was observed in all the genotypes. In crisis and steady state, ALT activity in HbSS increased with age (11–20 years) in steady state. However, with the age range studied there was no significant difference (p ≥ 0.05) in mean enzyme activity values when compared with normals. The pattern could change in older subjects, but for age 0–25 years at least hepatic dysfunction based on this enzyme alone could not be definitely deduced (see tables 3.1, 3.2.1).

Table 3.1: Effects of age on ALT, AST, and ALP activities in HbAA, HbAS and HbSS subjects.

Age (yrs)

ALT Activty (µL) AS

0-5 n=5

5.3 ± 1.0

5.5 ± 1.2

30.1 ± 2.0

7.5 ± 0.2

15.75 ± 0.4

19.75 ± 0.6

137.66 ± 23.1

102.8 ± 18.2

148.75 ± 14.9

6-10 n=5

11.7 ± 0.8

13.2 ± 0.9

36.9 ±1.8

13.33 ± 0.4

14.70 ± 0.3

10.5 ± 0.5

100.33 ± 15.2

102.8 ± 12.6

126.0 ± 8.49

11-15 n=5 16-20 n=5 21-25 n=5 >25 n=5

7.8 v 1.2

7.5 ± 1.0

26.8 ± 1.4

5.5 ± 0.2

11.0 ± 0.1

31.0 ± 1.9

66.75 ± 6.7

70.0 ± 5.6

88.7 ± 26.6

7.2 ± 0.6

10.5 ± 0.8

43.5 ± 1.2

9.67 ± 0.7

14.5 ± 0.2

39.1 ± 2.0

40.8 ± 13.3

49.5 ± 0.71

72.4 ± 13.6

8.0 ± 1.0

7.5 ± 0.9

12.0 ± 0.2

13.5 ± 0.4

31.5 ± 0.7

28.0 ± 2.8

8.0 ± 1.0

8.8 ± 1.0

10.25 ± 0.7

10.67 ± 0.5

29.8 ± 2.4

31.0 ± 16.2

AST Activty (µL) AS

n = number of samples analysed. Values are Mean ± SD.

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ALP Activty (µL) AS

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Table 3.2.1: Effect of age on ALT activity in HbSS subjects (Crisis and steady state).

Steady state Age (yrs) 15.3 ± 0.5 0–5 n=5 15.3 ± 0.6 6–10 n=4 23.7 ± 0.4 11–15 n=4 28.4 ± 0.2 16–20 n=5 n = number of samples analysed. Values are Mean ± SD.

ALT Activity (µL) Age (yrs) 0–5 n=3 6–10 n=3 11–15 n=3 16–20 n=3

Crisis 30.0 ± 0.7 36.8 ± 1.4 26.8 ± 0.4 43.5 ± 0.8

Table 3.2.2: Effect of age on AST activity in HbSS subjects (Crisis and steady state). AST Activity (µL) Steady state 19.75 ± 0.6

Age (yrs) 0–5 n=5 10.5 ± 0.5 6–10 n=5 31.0 ± 0.4 11–15 n=5 33.2 ± 0.6 16–20 n=5 n = number of samples analysed. Values are Mean ± SD.

Crisis 30.0 ± 0.7 36.8 ± 1.4 26.8 ± 0.4 43.5 ± 0.8

Table 3.2.3: Effect of age on ALP activity in HbSS subjects (Crisis and steady state). ALP Activity (µL) Steady state 148.75 ± 14.9

Age (yrs) 0–5 n=5 126.0 ± 8.49 6–10 n=5 88.7 ± 26.6 11–15 n=5 72.4 ± 13.6 16–20 n=5 n = number of samples analysed. Values are Mean ± SD.

Crisis 164.4 ± 4.3 155.6 ± 7.6 150.0 ± 9.9 119.0 ± 4.4

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Table 3.3.1: Effect of sex on ALT, AST and ALP activities in HbAA and HbAS subjects ALT Activity AST Activity AA AS Sex AA AS Sex 6.8 ± 7.6 ± Males 9.0 ± 13.0 ± Males 0.4 0.4 n=5 4.7 4.0 n = 15 8.0 ± Females 10.0 ± 12.0 ± Females 9.0 ± 0.52 0.4 n=9 6.85 5.6 n=9 n = number of samples analysed. Values are Mean ± SD.

Sex Males n=5 Females n=9

ALP Activity AA AS 57.5 ± 68.0 ± 3.9 5.9 56.8 ± 62.0 ± 2.9 4.6

SS 125.6 ± 3.8 121.3 ± 2.9

Table 3.3.2: Effect of sex on AST and ALP activity in HbSS subjects (Crisis and steady state). AST Activity Steady State Sex Sex 23.8 ± Males Males 13.54 n=8 n=5 23.5 ± Females Females 12.18 n=7 n=5 n = number of samples analysed. Values are Mean ± SD.

Crisis 43.88 ± 11.56 35.14 ± 6.36

Aspartate tarnsaminase (AST): The level of AST activity in sickle cell anaemia (HbSS) subjects was higher than normal. In HbAS there was a slight decrease in activity as age increased. For HbSS subjects, AST activity was high at age 15. Subjects in the age bracket 6–10 years exhibited a decrease in AST activity when compared to those between 0–5 years. In steady and crisis situation, AST activity was highest in HbSS subjects when compared to HbAA and HbAS subjects. However, during crisis a higher activity was observed for all age groups studied which appears to be no age dependent trend in AST activity for HbSS subjects under crisis situation with mean values for each age group increase progressively (see table 3.2.2). AST activity in steady state increased with age up to age 20. Subjects in steady state between 6–10 years exhibited a low AST activity when compared to subjects between 0–5 years. However, there was no significant difference (p ≥ 0.05) in AST activity

Sex Males n=5 Females n=5

ALP Activity Steady state Crisis 125.6 ± 3.8 144.6 ± 2.7 10.0 ± 6.85

157.3 ± 7.8

in HbSS subjects between 11–15 years and 16– 20 years (see table 3.1). On the effect of sex, statistical analysis showed no significant difference (p≥ 0.05) between both sexes in AST activity for HbAA, HbAS and HbSS (crisis and steady state) subjects. Alkaline phosphatase (ALP): From table 3.1 above, ALP activity of heterozygous (AS) is not significantly different (p ≥ 0.05) from that of homozygous (AA) subjects. As shown in table 3.2.3, ALP activity increased during crisis compared to steady state condition. The effect of ALP activity in normal (AA) and heterozygous (AS) subjects decreased with age significantly (p ≤ 0.05), while a steady decrease in ALP activity was observed with increase in age for HbSS subjects. However, the difference in activity between the various age groups is statistically significant (p ≤ 0.05).

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Table 3.3.1 shows the effect of sex on ALP activity of normal and sickle cell subjects. The mean value for each group did not depend on sex. There was no significant difference (p ≥ 0.05) in ALP activity between both sexes.

calcification of bone and teeth. Also, raised serum alkaline phosphatase accompanies rickets of various etiologies.

Table 3.3.2 shows the effect of sex on ALP activity in HbSS subjects in steady state and painful crisis. Sex did not seem to affect the enzyme activity whether in steady state or crisis.

In conclusion, the ALT and AST activities in sickle cell disease as regards age are raised from childhood to adolescence and that in the steady state of sickle cell, there were elevations which were further increased during painful crisis. The sexes may not lead to any major hepatic dysfunction, but should be considered as a diagnostic parameter in sickle cell management.

In this work, ALP had the highest levels of activity, especially in growing children – in both normal and sickle cell subjects. This may be as a result of its involvement in the

CONCLUSION

REFERENCES Acquaye, C., Wilchek, M. and Gorecki, M. (1981). Strategies for tackling sickle cell disease. Trends in Biochemical Sciences, 146–148. Attah, E.B. (1975). Pathology of sickle cell anaemia. Dokita. 7:19–21. Dacie, J.V. (1960). The haemolytic anaemia, congenital and acquired anaemia. (2nd ed.). J.A. Churchill Ltd. Part 1. London: 200–230. Dean, J. and Schechter, A.N. (1978). Sickle cell anaemia. Molecular and cellular basis of therapeutic approaches. The New England Journal of Medicine. 229:753– 755. Ekeke, G.I. and Ibeh, G.O. (1990). Liver function enzymes and serum inorganic phosphate levels in sickle cell disease. 2: 6–9.

Emmel, V.E. (1917). A study of the erythrocyte in the case of severe anaemia with elongated and sickle shape red blood corpuscles. Arch. Intern. Med. 20: 586. Hahn, E.V. and Gillepspie, G.E.G. (1927). Sickle cell anaemia. Arch. Intern. Med. 39: 233–236. Herrick, J.B. (1910). Peculiar elongated and sickle shaped red blood corpuscle in a state of severe anaemia. Arch. Intern. Med. 6: 517–521. Konotey-Ahulu, F.I.D. (1974). The sickle cell diseases, clinical manifestations including the “sickle crisis”. Arch. Intern. Med. 133: 611–619. Lehmann, H. and Raper, A.B. (1949). Distribution of sickle cell trait in Uganda, and its ethnological significance. Nature, 164: 494–495.

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Nygren, A. (1967). SGOT in chronic alcoholism. Act. Med. Scand. 182–383.

Sofowora, A.O. (1993). Medicinal plants and traditional medicine in Africa. 2nd ed. Ibadan: Spectrum books limited; pp 150.

Smith, C.H. (1972). Blood disease of infancy and childhood (3rd ed.). C.V. mosby and Co., St. Louis, 353–449.

White, A., Handler, P., Smith, E.L., Hill, R.L. and Lehmann, I.R. (1978). Principles of Biochem., 6th ed. 947–1003.

Source of Support: Nil

Conflict of Interest: None Declared

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Review article CHELIDONIUM MAJUS L. - A REVIEW ON PHARMACOLOGICAL ACTIVITIES AND CLINICAL EFFECTS Biswas Surjyo Jyoti1* 1

Department of Zoology, Midnapore College, Midnapore, West Bengal, India-721101 *Corresponding Author: E-mail: surjyo@rediffmail.com

Received: 01/03/2013; Revised: 25/03/2013; Accepted: 30/03/ 2013

ABSTRACT Chelidonium majus L. (Papaveraceae) is a plant that has been used for centuries in treating many diseases in European and Asian countries. Crude extracts from various parts of the plant contain isoquinoline alkaloids. The alkaloids derived from C. majus have not yet much studied; however, some reports are available on toxicity studies of alkaloids of this plant. In such a scenario there is need for understanding its therapeutic potential and its toxic actions. This review summarizes scientific findings and suggests areas where further research is needed. KEY WORDS: Chelidonium majus, alkaloids, pharmacology, antioxidant

Cite this article: Biswas S J (2013), CHELIDONIUM MAJUS: A REVIEW ON PHARMACOLOGICAL ACTIVITIES AND CLINICAL EFFECTS., Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 238–245

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INTRODUCTION:

Uses in traditional medicine systems

Chelidonium majus commonly known as swallow-wort, rock poppy or greater celandine belongs to Family-Papaveraceae. This plant is distributed across the globe viz. Europe, Asia, North America and in northwest Africa, particularly in soils rich in nitrogen. The name â&#x20AC;&#x2DC;Chelidoniumâ&#x20AC;&#x2122; came from Chelidon-a greek word which means swallow bird, as the plant begins to flower when the swallows return. The plant is widely regarded for its therapeutic potential in Western and Asian countries particularly in Chinese traditional medicine and homeopathy. Crude extracts of C. majus and isolated compounds exhibit numerous biological activities (Colombo and Bosisio, 1996; Gilca et al., 2010). Though many diseased conditions even today are being treated with C. majus both in traditional and homeopathic medical systems but it has some self limitations therefore, its therapeutic efficacy needs critical evaluation. The current review summarizes scientific findings of other investigators on C. majus and suggests areas where further investigations/research is needed.

In many European, Asian and African countries C. majus latex was used for bile and liver disorders, for treatment of warts, corns, eczema and solid tumors. It has traditionally being used to treat liver diseases, gastric ulcer, tuberculosis, skin eruptions and oral infections. In Chinese traditional medicine and in homeopathy C. majus is used to treat blockage of blood circulation, to relieve pain edema and jaundice. Phyto-constituents (Figure. 1) Extracts of Chelidonium has been found to contain three types of benzyl isoquinoline alkaloids viz. protoberberine, protopine, benzophenanthredine. Sanuinarine and chelerythrine are the prominent compounds obtained from roots while coptisine, chelidonine and berberine are obtained from the aerial parts (Colombo and Bosisio, 1996). Other constituents include malic, citric, gentisic, and hydrobenzoic acids. It also contains hydroxycinnamic acid derivatives, sparteine, saponin, carotenoids, chelidocystatin and flavonoids.

Figure. 1 Chemical structure of Phyto-constituents

PHARMACOLOGICAL ACTIVITIES Hepatoprotective effects It has been demonstrated that Chelidonium majus favourably modulates carbon tetrachloride induced toxicity in rats. The treatment with C. majus considerably reduced the number of necrotic cells and decreased the activities of transaminases and bilirubin (Mitra et al., 1992; Mitra et al., 1996). Biswas et al. (2008) have

reported that ethanolic whole plant extract of Chelidonium majus, has been tested for its possible anti-tumor, hepato-protective and antigenotoxic effects in p-dimethylaminoazobenzene (p-DAB) induced hepatocarcinogenesis in mice through multiple assays: cytogenetical, biochemical, histological and electron microscopical. Data of several cytogenetical endpoints and biochemical assay of some toxicity marker enzymes at all fixation intervals and histology of liver sections through ordinary,

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scanning and transmission electron microscopy at certain fixation intervals were critically analyzed. The results suggest anti-tumor, antigenotoxic and hepato-protective effects of the plant extract, showing potentials for use in cancer therapy. Chung et al., (2004) demonstrated that C. majus enhances nitric oxide and TNF-α production via NF kappa B activation in mouse. Effects on enzymes Mazzanti et al., (2009) reported that there was a significant reduction in glutathione level and SOD activity in liver after high oral dose of C. majus. It was reported by others that C. majus has a strong antioxidant activity as revealed from FRAP assay (Then et al., 2003). Biswas et al., (2008) also reported that LPO and transaminases activity reduced significantly after treatment with C. majus extract against p-DAB induced hepatocarcinogenesis. Antimicrobial, antiviral and antiparasitic effects

Alkaloids extract showed antiviral efficacy against human adenoviruses type 5 and 12, herpes simplex virus, and RNA polio virus (Zuo et al., 2008; Horvath et al., 1983; Kery et al., 1987). Zhu and Ahrens (1982) investigated that berberine successfully controlled the intestinal secretion enhanced by E. coli enterotoxin, the effect of which was dose dependent and it may be due to quaternary ammonium group which is responsible for anti-bactericidal property of berberine and protoberberine was found active against reverse transcriptase enzyme of RNA tumor viruses. Chelidocystatin decreases the activity of cysteine proteinases but further in depth research are necessary especially in vivo conditions. Cardiovascular effects Sanguinarine has been involved in suppression of angiogenesis by inhibition of VEGF signaling, this has been experimentally proved in pig granulosa cells and in porcine endothelial cells (Basini et al., 2007). Immuno-modulatory activity

The modulatory effect of C. majus extract against virus was evaluated in various in vitro and in vivo studies C. majus showed antimicrobial effect on gram positive bacteria and on Candida albicans (Lendfeld et al., 1981). Crude extracts of several alkaloids extracted from C. majus exhibited antimicrobial, antiviral and antifungal properties (Lozyuk, 1977; Gerencer, et al., 2006; Parvu et al., 2008; Meng et al., 2009; Monavari et al., 2012). Growth of Alternaria, Aspergillus flavus, Candida albicans, Rhizopus orizae and Scopulariopsis was inhibited by berberine at 10–25 μg/ml concentration (Mahajan et al., 1982). Ma et al., 2000 demonstrated that chelidonine, dihydrochelerythrine and dihydrosanguinarine isolated from C. majus roots have activity against Cladosporium herbarum at 4-10 μg/ml concentration. It has been experimentally proved that compounds (8-hydroxydihydrosanguinarine, dihydro-sanguinarine, dihydrochelerythrine, 8-hydroxydihydro-chelerythrine) isolated from aerial parts of the plant showed anti-bacterial effect against methicillin resistant Staphylococcus aureus (Zuo et al., 2008).

Immuno-modulatory properties of C. majus have been investigated by Song et al., 2002, where he obtained a protein bound to polysaccharide from water extracts of the plant, (CM-Ala) which showed mitogenic activity on spleen, bone marrow cells, it also increased the number of granulocyte macrophage colony forming cells, further it suppressed immune response locally by decreasing epidermal Langerhans cells (Song et al., 2002). It has been demonstrated that C. majus extract improved overall humoral and cellular immunity response and decreased incidence of recurrences of tonsillitis in children (Khmel’nitskaia et al., 1998). Anti-inflammatory and Analgesic activity Stylopine is a major component of leaf of C. majus and it suppresses NO and PGE2 production in macrophages by inhibiting iNOS and COX 2 expressions. It has been demonstrated that 5 and 12 lipoxygenase were inhibited by sanguinarine and chelerythrine because these enzymes are

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involved in leukotriene B4 and 12 hydroxyeicosatetranoic acid syntheses. As compared to chelerythrine, sanguinarine showed higher anti-inflammatory activity due to different oxygen electron donating constituents (Lendfeld et al., 1981). It has been reported that C. majus extract increases TNF α production due to NF κB production. It has also been reported that Ukrain induces depolarization of mitochondrial membrane potential and activates caspase in Jurkat T lymphoma cell model (Habermehl et al., 2006).

(2011) reported that berberine inhibits mitochondria function and decreases intracellular ATP in streptozotocin induced diabetes in rats. This leads to a reduction in transcription factors such as FoxO1, SREBP1, and ChREBP. As a result, expression of gluconeogenic genes (PEPCK and G6Pase) and lipogenic gene (FAS) decreases. These molecular changes represent a signaling pathway for improvement of fasting glucose in the berberine treated diabetic rats (Xia et al., 2011). Anti-cancer efficacy

Choleretic effects Vahlensieck et al., (1995) used phenolic and alkaloid fractions of C. majus for their choleretic activity using perfused rat livers. He demonstrated that total extract induced choleresis i.e. the bile flow was significantly elevated and the amount of the bile was more than double the quantity. Though it was not ascertained which fraction of the extract was responsible for the increased bile flow. Effects on reproductive systems The feeding of ethanolic extract of C. majus showed that it could combat the spermatotoxic effects to some extent in induced p-DAB induced carcinogenesis. As benzophenanthridine alkaloids have marked nucleophilic properties, they might intercept the reactive metabolites; thereby preventing their attack on nucleophilic sites on DNA, and hence blocking adduct formation (Vavreckova et al., 1996 a, b). Further it has been suggested that many enzymatic functions are essential for the normal integrity and function of testis i.e. synthesis, development and maintenance of normal sperm. Therefore, the protective role of C. majus on sperm head could also be attributed to its regulatory effect on protein metabolism and repair activities in the germinal cells (Biswas and Khuda-Bukhsh, 2002). Antihyperglycemic and Hypoglycemic activity Berberine an isoquinoline alkaloid obtained from C. majus is used widely in China to reduce blood glucose, in type II diabetes. Xuan et al.,

The anti-leukaemic activity of protoberberine alkaloids has been reported and Smekal et al. (1984) demonstrated that sanguinarine intercalates partially as well as totally into the DNA double helix. It has been demonstrated by circular dichroism that the spectrum of DNA is similar to ethidium binding to DNA. C. majus had antiproliferative effect on human keratinocyte cell lines (Vavreckova et al., 1996a, b). Berberine intercalation to DNA might be due to the planes of intercalated molecules which lie parallel to those of purine-pyrimidine pairs. An important constituent berberine has been shown to interact with nucleic acids by various optical methods. It was tested that administration of 350 μg/kg of protopine intra-peritoneally inhibited very less regression of Erlich carcinoma and application of 50 μg/kg b.w. of chelidonine regressed sarcoma 180 (Sokoloff, 1968). UkrainTM an anticancer drug whose components include most of C. majus compounds exerts multiple effects on cancer cell lines (Cordes et al., 2003). Several reports have been obtained in both animal and human models regarding anticancer efficacy of Ukrain, against various types of induced stomach carcinogenesis, induced hepatocarcinogenesis, in patients suffering from pancreatic cancer, Kaposis sarcoma (Kim et al., 1997; Biswas and KhudaBukhsh, 2002; Lohninger et al., 1996; Gansauge et al., 2002; Ernst and Schmidt, 2005). Chelidonine inhibits telomerase in tumor cells strongly and may provide a basis for probable anticancer agent also this alkaloid arrest mitosis as a result of inhibition of tubulin polymerization and activation of SAPK/JNK pathway.

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Central Nervous system

Cytotoxic effects

An alkaloid obtained from C. majus, thiophosphoric acid has been tested on rodents regarding it action on CNS, it was found that it depresses spontaneous motor activity; it seems to stimulate dopaminergic system and depresses the serotoninergic system (Kleinrok et al., 1992).

There were spontaneous reports of adverse drug reactions associated with C. majus preparation. Incidences of hepatotoxicity have been reported by several authors (Moro et al., 2002, Kaminsky et al., 2006). It has been reported that C. majus showed cytotoxicity towards lymphoma cells and murine cell lines. In some countries Complementary Evaluation Committee recommended that products containing alkaloids obtained from C. majus must have a warning label and it should be administered under medical supervision only.

CLINICAL STUDIES: Dysentry or gastroenteritis Ardjah (1991) studied action of celandine on upper abdominal symptoms in human subjects such as cholinergic and spasmolytic effects using panchelidon®. In case of patients with postcholecystectomy 29 patients out of 35 showed clear improvement. A similar study using 21 patients with dyspeptic complaints with alcohol toxic liver parenchyma damage, 20 reported improvement after two weeks of treatment. Limited numbers of clinical studies have been carried out with total extracts in patients with epigastric complications and the sample size was small and definite conclusion could not be ascertained from the study. Periodontal effects Benzophenanthridine alkaloids are routinely used for the treatment of periodontal diseases, Boulware et al., (1985) investigated that sanguinaria extract was helpful in lowering of volatile sulphur present in the oral cavity. Southard et al., (1987) reported that benzophenanthridine alkaloids act as an anticaries thereby preventing tooth decay. Radioprotective effects

CONCLUSION We have reported hepatoprotective ability of C. majus crude extract and various potencies of it in induced hepatocarcinogenesis. It would be prudent to investigate its constituents singly and in combination, how they modulate pathological changes and which form is more potent or effective. Time of collection of plant materials, place of collection, extraction procedures, and its storage might affect its active compounds both quantitatively and qualitatively. The information summarizes here concerning C. majus is intended to serve as a reference to researchers involved in ethnopharmacological research. ACKNOWLEDGEMENT Grateful acknowledgements are made to Professor A. R Khuda-Bukhsh, Department of Zoology, University of Kalyani and Dr. Prabir De, Scientist, CCMB, Hyderabad for encouragements.

Song et al., 2003 demonstrated that extracts of C majus have certain radioprotective effects.

REFERENCES Amoros M, Fauconnier B, Girre L (1977). Propriétés antivirales de quelques extraits de plantes indigenes. Annals Pharm Françaises. 35:371–376.

Ardah H (1991). Therapeutic aspects of functional symptoms of the upper abdomen in biliary tract diseases Fortschr Med.Med. 109 (S 115): 2–8.

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Basini G, Santini S, Bussolati S, Grasselli F (2007). The plant alkaloid sanguinarine is a potential inhibitor of follicular angiogenesis. J. Reproduc. Dev. 53(3):573–79. Biswas SJ, Bhattacharjee N, Khuda Bukhsh AR (2008). Efficacy of a plant extract (Chelidonium majus L) in combating induced hepatocarcinogenesis in mice. Food Chem. Toxicol 46(5):1474–87. Biswas SJ, Khuda-Bukhsh AR (2002). Effect of a homeopathic drug, Chelidonium, in amelioration of p-DAB induced hepatocarcinogenesis in mice. BMC Complement. Altern. Med. Vol. 2/4: 1– 16. Boulware RT, Southard GL, Yankell SL (1985). Sanguinaria extract, a new agent for the control of volatile sulphur compounds in the oral cavity. J Soc. Cosmetic Chem. 36(4):297–302 Chung HS, An HJ, Jeong HJ, Won JH, Hong SH, Kim HM (2004). Water extract isolated from Chelidonium majus enhances nitric oxide and tumor necrosis factor alpha production via nuclear factor kappa B activation in mouse peritoneal macrophages. J. Pham. Pharmacol. 56(1): 129–134. Colombo M, Bosisio E (1996). Pharmacological activities of Chelidonium majus L. (Papaveraceae). Pharmacol Res. 33(2): 127–134 Cordes N, Blaese MA, Plasswilm L, Rodemann HP, Van Beuningen D (2003). Fibronectin and laminin increase resistance to ionizing radiation and the cytotoxic drug Ukrain in human tumour and normal cells in vitro. Int. J. Radiat. Biol. 79:709–20. Ernst E, Schmidt K (2005). Ukrain – a new cancer cure? A systematic review of randomized clinical trials. BMC Cancer 5: 69.

Gansauge F, Ramadani M, Pressmar J, Gansauge S, Muehling B, Stecker K, Cammerer G, Leder G, Beger HG (2002). NSC-631570 (Ukrain) in the palliative treatment of pancreatic cancer. Results of a phase II trial. Langenbecks Arch. Surg. 386:570–4. Gerencer M, Turecek PL, Kistner O, Mitterer A, Savidis-Dacho H, Barrett NP (2006). In vitro and in vivo anti-retroviral activity of the substance purified from the aqueous extract of Chelidonium majus L. Antiviral Res. 72 (2): 153–6. Gilca ML, Gaman E, Panait I, Stoian, Atanasiu V (2010). Chelidonium majus- an intergrative review: Traditional knowledge versus modern findings. Forsch Komplementmed 17(5): 241–8. Habermehl D, Kammerer B, Handrick R, Eldh T, Gruber C, Cordes N, Daniel PT, Plasswilm L, Bamberg M, Belka C, Jendrossek V (2006). Proapoptotic activity of Ukrain is based on Chelidonium majus L alkaloids and mediated via a mitochondrial death pathway. BMC Cancer. 6:14. Horvath J (1983). Antiviral effect of Chelidonium extracts. Proc. Int. Congr. Chemother. 9:124/106–124/110. Kaminsky V, Lootsik M, Stoika R (2006). Correlation of the cytotoxic activity of four different alkaloids, from Chelidonium majus (greater celandine), with their DNA intercalating properties and ability to induce breaks in the DNA of NK/Ly murine lymphoma cells. Cent. Eur. J. Biol. 191: 2–15. Kery RY, Horvath J, Nasz I, Verzar-Petri G, Kulcsar G, Dan P (1987). Antiviral alkaloid in Chelidonium majus L. Acta Pharmaceu. Hungerica. 57: 19–25.

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Khmel’nitskaia NM, Vorob’ev KV, Kliachko LL, Ankhimova ES, Kosenko VA, Tymova EV, Mal’seva GS, Medvedev EA (1998). A comparative study of conservative treatment schemes in chronic tonsillitis in children. Vest. Otorinolaringol. 4: 39–42. Kim DJ, Ahn. B, Han BS, Tsuda H. (1997). Potential preventive effects of Chelidonium majus L. (Papaveraceae) herb extract on glandular stomach tumor development in rats treated with N-methyl- N0-nitro-N-nitrosoguanidine (MNNG) and hypertonic sodium chloride. Cancer Lett. 112: 203–8. Kleinrok Z, Jagiello-Wojtowicz E, Matuszek B, Chodkowska A. (1992). A basic central pharmacological properties of thiophosphoric acid alkaloid derivatives from Chelidonium majus L. Pol. J. Pharmacol. Pharm. 44: 227–239. Lendfeld J, Kroutil M, Marsalek E, Slavik J, Preininger V, Simanek V (1981). Isolation, chemistry and biology of alkaloids from plants of the papaveraceae: anti-inflammatory activity of quaternary benzophenanthridine alkaloids from Chelidonium majus. Planta Medica 43(2): 161–5. Lohninger A, Korsh OB, Melnyk A (1996). Combined therapy with Ukrain and chemotherapy in ovarian cancer (case report). Drugs Exp. Clin. Res. 22: 259– 62. Lozyuk LV (1977). Antiviral properties of some compounds of plant origin. Mikrobiol. Zh (Kiev). 39: 343–8. Ma WG, Fukushi Y, Tahara S, Osawa T (2000). Fungitoxic alkaloids from Hokkaido Papaveraceae. Fitoterapia. 71(5): 527–34.

Mahajan VM. (1982). Antimycotic activity of berberine sulphate: an alkaloid from an Indian Medicinal herb. Sabouraudia 20: 79–81. Mazzanti G, Sotto A Di, Franchitto A, Mammola CL, Mariani P, Mastrangelo S, Menniti-Ippolito F, Vitalone A. (2009). Chelidonium majus is not hepatotoxic in Wistar rats, in a 4 weeks feeding experiment. J. Ethnopharmacol. 126 (3): 518–24. Meng F, Zuo G, Hao X, Wang G, Xiao H, Zhang J, Xu G (2009). Antifungal activity of the benzo[c]phenanthridine alkaloids from Chelidonium majus Linn against resistant clinical yeast isolates. J. Ethnopharmacol. 125(3): 494–6. Mitra S, Gole M, Samajdar K, Sur RK, Chakraborty BN (1992) Antihepatotoxic activity of Chelidonium majus. Int J Pharmacognosy 30:125–8. Mitra S, Sur RK, Roy A, Mukherjee AS (1996). Effect of Chelidonium majus L. on experimental hepatic tissue injury. Phytother Res. 10: 354–356. Monavari SH, Shahrabadi MS, Keyvani H, Bokharaei-Salim F (2012). Evaluation of in vitro antiviral activity of Chelidonium majus L. against herpes simplex virus type-1. African. J. Microbiol. Res. 6(20): 4360–64. Moro PA, Cassetti F, Giugliano G, Falce MT, Mazzanti G, Menniti-Ippolito F,Raschetti R, Santuccio C (2009).Hepatitis from Greater celandine (Chelidonium majusL.): review of literature and report of a new case. J Ethnopharmacol.124(2):328–32.

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Parvu M, Parvu AE, Cranium C, BarbuTudoran L, Tamas M (2008). Antifungal activities of Chelidonium majus extract on Botrytis cinerea in vitro and ultrastructural changes in its conidia. J. Phytopath. 156(9): 550–2.

Vahlensieck U, Hahn R, Winterhoff H, Gumbinger HG, Nahrstedt A, Kemper FH (1995). The effect of Chelidonium majus herb extract on choleresis in the isolated perfused rat liver. Planta Med. 61:267–271.

Smekal E, Kubova N, Kleinwachter V (1984). Interaction of benzophenanthridiene alkaloid sanguinarine with DNA. Stud. Biophys. 101:125–32.

Vavreckova C, Gawlik I, Muller K.1996b. Benzophenanthridine alkaloids of Chelidonium majus: 2. Potent inhibitory action against the growth of human keratinocytes. Planta Med. 62 (6):491– 4.

Sokoloff B (1968). Oncostatic and oncolytic factors present in certain plants. Oncology 22(1): 49–60. Song JY, Yang HO, Pyo SN, Jung IS, Yi SY, Yun YS. (2002). Immunomodulatory activity of protein-bound polysaccharide extracted from Chelidonium majus. Archives. Pharmacol. Res. 25(2):158–164. Song JY, Yang HO, Shim JY, Ji-Yeon-Ahn, Han YS, Jung IS, Yun YS (2003). Radiation protective effect of an extract from Chelidonium majus. Int. J. Hematol. 78(3):226–32. Southard GL, Parsons LG, Thomas LG, Woodall IR, Jones BJ. (1987) Effect of sanguinaria extract on development of plaque and gingivitis when supragingivally delivered as a manual rinse or under pressure in an oral irrigator. J. Clin. Periodontol. 14(7): 377–80. Then M, Szentmihalyi K, Sarkozi A, Varga IS (2003). Examination of antioxidant activity in greater celandine (Chelidonium majus L.) extracts by FRAP method. Acta Biol. Szeged 4791– (4):115–7.

Source of Support: Nil

Vavreckova C, Gawlik I, Muller K. 1996a. Benzophenanthridine alkaloids of Chelidonium majus:1 Inhibition of 5and 12-lipoxygenase by a non redox mehanism. Planta Med. 62 (5):397–401. Xia X, Yan J, Shen Y, Tang K, Yin J, Zhang Y, Yang D, Liang H, Ye J, Weng J (2011). Berberine improves glucose metabolism in diabetic rats by inhibition of hepatic gluconeogenesis. PLoS One. 3; 6(2):e16556. doi:10.1371/journal.pone.0016556. Zhu B, Ahrens FA (1982). Effect of berberine on intestinal secretion mediated by Escherichia-coli heat stable enterotoxin in jejunum of pigs. American J. Vet. Res. 43(9):1594–8. Zuo GY, Meng FY,. Hao XY, Zhang YL, Wang GCH,. Xu. GL (2008). Antibacterial alkaloids from Chelidonium majus Linn (Papaveraceae) against clinical isolates of methicillin-resistant Staphylococcus aureaus. J. Pharm. Pharmaceut. Sci. 11(4): 90–4.

Conflict of Interest: None Declared

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Research article A DETAILED PHARMACOGNOSTICAL EVALUATION ON LEAF OF OLAX SCANDENS ROXB. Naik Raghavendra1*, Borkar Sneha D2, Harisha C R3, Acharya R N4 1,2

P G Scholar, Department of Dravyaguna, IPGT & RA, Gujarat Ayurved University, Jamnagar, Gujarat, India 3 Head, Pharmacognosy Laboratory, IPGT & RA, Gujarat Ayurved University, Jamnagar, Gujarat, India 4 Associate professor, Department of Dravyaguna, IPGT & RA, Gujarat Ayurved University, Jamnagar, Gujarat, India *Corresponding Author: Email: ayuraghu@gmail.com

Received: 11/02/2013; Revised: 22/02/2013; Accepted: 28/03/ 2013

ABSTRACT Leaves of Olax scandens Roxb. (Olacaceae) are edible and used for cure of headache. A detailed pharmacognostical character of its leaf is lacking. In the present study, its leaves evaluated for their morphological, microscopical and quantitative microscopic characters following standard procedures. The overall study showed the leaves are 3.5–9 × 2.5–3.2 cm in size, reticulate venation, petiole 0.2–0.5 cm. Leaves showed the presence of trichomes, collenchyma, and vascular bundles. Powder microscopy of the dried leaves shows paracytic stomata of lower epidermis, epidermal cells of upper epidermis, and unicellular trichomes of epidermis, rosette and prismatic crystals of calcium oxalate. KEY WORDS: Olax scandens, microscopy, quantitative methods, stomata.

Cite this article: Naik R, Borkar S D, Harisha C R, Acharya R N (2013), A DETAILED PHARMACOGNOSTICAL EVALUATION ON LEAF OF OLAX SCANDENS ROXB., Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 246–253

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INTRODUCTION

Morphological study.

Plant and various plant products are being used by human and animals either directly or indirectly, since the existence of life, for food, medicine, clothing, shelter etc. Olax scandens Roxb. (Olacaceae), known as Badru, is a shrub or small tree, distributed throughout tropical India Flowers white, 6–7.5 mm long, in short racemes; Fruit yellow, or orange fleshy, subglobose, 0.8–1.5 cm in diameter, more than half enclosed in accresent calyx. (Saxena H.O, 1995). Leaves of Olax scandens Roxb. (Olacaceae) is being used by tribal people of Odisha for medicinal and food purposes (Tribhubana Panda et al., 2007). Decoction of stem bark is taken internally to cure fever and cough, (Veeramuthu et al., 2006) (Kirtikar & Basu, 2003), boiled leaves fomentation is applied to cure headache (Anonymous, 1990). Review of literature reveals that its leaves have not been studied in detail for pharmacognostical characters, which is an essential parameter for identification of a crude drug (Anonymous, 1999). Hence, the present study was undertaken to establish certain botanical standards for identification and standardization of O. scandens leaf.

The morphological study includes size, shape, apex, margin, venation, base, petiole, surface, color of leaves of O. scandens.

MATERIALS AND METHODS:

Macroscopic investigation showed that the leaves are alternate, exstipulate, petiolate, petiole 0.2–0.5 cm, somewhat twisted, base equal, leaf measures about 3.5–9 × 2.5–3.2 cm, dark green above, light green below, ovate to lanceolate, margin simple, apex obtuse, strong midrib, 6–7 pairs of nerves with reticulate venation. (Plate A1–2).

Collection and preservation of the sample Leaves of Olax scandens Roxb. were collected from its natural habitat, Balangir, Odisha, during September 2012 and identified with the help of botanical texts and flora (Saxena H.O, 1995). A sample specimen was deposited to Pharmacognosy lab (SPECIMEN NO- PHM 6062/21/09/2012) for future references. The leaves were washed, shade dried, powdered, sieved through 80 no. mesh and preserved in an air-tight glass vessel. For microscopical evaluation, fresh sample was preserved in a solution prepared from 70% ethyl alcohol : glacial acetic acid : formalin (AAF) in the ratio of 90:5:5 (Johnson Alexander Donald, 1940).

Microscopical study: Microscopical examinations were carried out by taking transverse section of petiole, leaf through midrib (Khandelwal K.R et al., 1996), type and distribution of stomata, epidermal cell and trichomes (Anonymous, 1999) following standard guidelines. Quantitative microscopy: Quantitative microscopy was carried out to determine epidermal cell number, stomatal number, stomatal index and size of the stomata (Wallis, 1985). Powder microscopy: Dried leaf powder was studied following standard procedures (Trease GE et al., 2002). The micro photographs were taken by using Carl zeiss trinocular microscope. RESULT AND DISCUSSION: Morphological study:

Microscopical study: Petiole: The T.S of petiole is half-moon shaped in outline. It showed, outer single layer of epidermis with numerous simple, horn shaped trichomes, followed by cortex, endodermis, pericyclic fibers, phloem and centrally located xylem. (Plate A 3–8).

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PLATE A

1. Morphology of leaf

3. T.S through petiole

2. Measurement of leaf

4. Rosette crystal & tannin content

5. Epidermal layer with trichomes and rosette crystal

Prismatic crystals & tannin

7. T.S showing cortex and vascular bundle

8. Pericyclic fiber, phloem and xylem.

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Epidermis is single layered, barrel shaped, compactly arranged cells, filled with yellow colored material. Some of the epidermal cells showed simple, unicellular, horn shaped trichomes. Epidermis is covered with thick cuticle. Cortex is made up of 8–10 layers of loosely arranged parenchyma cells. Lower side of the parenchyma cells filled with rosette crystals, prismatic crystals of calcium oxalate and some tannin contents as compared to the upper side. Some of the cells contained oil globules. Endodermis is Inner to the cortex, single layered, somewhat elongated thin walled cells forming endodermis. Around the endodermis 4–6 layers of pericyclic fibers forming a ring like structure covering the vascular bundle. Vascular bundles are open and bi-collateral type. Phloem present around the xylem with some phloem fibers and sieve elements forming a ring like structure. The metaxylem facing towards lower epidermis and protoxylem facing towards upper epidermis. Xylem bundles were separated by uniserrate medullary rays along with xylem parenchyma and fibers. T.S Through mid rib The T. S of leaf showed upper and lower epidermis with mesophyll tissue having upper pallisade and lower spongy parenchyma cells. Section through midrib showed centrally located vascular bundle covered with ground tissue. On the lower side of the transverse section 1–3 layers of collenchymatous cells were present. (Plate B1–4) Epidermis was Single layered, barrel shaped epidermal cells both on upper and lower epidermis with unicellular trichomes. Epidermis was covered with cuticle. Trichomes were more in lower epidermis than upper epidermis. Stomata found only at the lower epidermis.

Mesophyll tissue was differentiated into two layers. Upper 1–2 layers of compactly arranged pallisade parenchyma with oil globules, and rich in chloroplast pigments. Rarely with some rosette crystals of calcium oxalate. Lower 3–5 layers of spongy parenchyma cells, loosely arranged with air spaces and loaded with prismatic, rosette crystals of calcium oxalate. Section through mid-rib showed a large vascular bundle located at the centre, 1–3 layers of collenchyma tissue present at the lower epidermis surrounding the ground tissue. Ground tissue was made up of thin walled compactly arranged parenchyma cells heavily loaded by rosette crystals, prismatic crystal of calcium oxalate and some oil globules. Inner to the ground tissue, single layered, somewhat elongated thin walled cells forming endodermis. Inner to the endodermis 4–5 layers of pericyclic fibers forming a ring like structure covering the vascular bundle. Vascular bundle was open and bi-collateral in type. Phloem’s present around the xylem with some phloem fibers and sieve elements formed a ring like structure. The metaxylem facing towards lower epidermis and protoxylem facing towards upper epidermis. The xylem bundles were separated by uniserrate medullary rays along with xylem parenchyma and fibers. Surface study of epidermis: (Plate B5–8) Surface study of epidermis was carried out to determine type and distribution of stomata, epidermal cell and trichomes. Stomata were absent in the upper epidermis and consists only wavy epidermal cells. Some of the trichomes and cicatrix were also observed. The lower epidermis consists of numerous stomata of paracytic type. Some of the trichomes and cicatrix were also observed.

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PLATE-B

1. T.S through mid rib

Prismatic crystals

3. Vascular bundles with ground tissue

Xylem and phloem

Lower epidermis with paracytic stomata

7. Upper epidermis with cicatrix

6. Stomata & cicatrix with epidermal cells

8. Upper epidermis with trichome

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PLATE- C

1. Lower epidermis with stomata

3. Simple trichome

2. Measurements of stomata

4. Crystal fibers

5. Prismatic crystal

6. Epidermal cells

7. Paracytic stomata

8. Annular & spiral vessels

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TABLE 1: Quantitative microscopy of O. scandens leaf Sr. No

Parameter

Result

Type of the stomata

Paracytic

Length of the stomata

29.99 μm

Width of the stomata

22.43 μm

Outline of the stomata

673.19 μm2

Number of the stomata

Number of epidermal cells

Stomatal index

Quantitative microscopy: (Plate C1–2)

CONCLUSION

Quantitative microscopy of leaves was carried out to determine epidermal cell number, stomatal number, stomatal index and size of the stomata.

Leaf of Olax scandens Roxb. (Olacaceae) can be identified on the basis of key microscopical characters like paracytic stomata, unicellular, simple horn shaped trichomes, rosette and prismatic crystals of calcium oxalate, bi collateral vascular bundles, annular and spiral vessels, tannin, crystal fibers and lignified fibers. The quantitative surface microscopy study showed 24 numbers of epidermal cells, and stomatal index 33. These observed parameters could be useful to establish certain botanical standards for identification and standardization of O. scandens leaf.

The stomatal number, stomatal index, stomatal size, epidermal cell size were calculated by trial and error method (by taking 3–5 successive readings. Mean value was taken into consideration.) Results are tabulated in table – 1 Powder microscopy (Plate C3–8) Powder microscopy of the dried leaf powder was carried out following standard guidelines. Organoleptic characters showed the presence of greenish color with leafy odour and bitter taste. Microscopic characters Diagnostic characters of powder microscopy showed the paracytic stomata from lower epidermis, epidermal cells of upper epidermis, unicellular, simple horn shaped trichomes from epidermis, rosette and prismatic crystals of calcium oxalate from mesophyll and ground tissue. Annular and spiral vessels from vascular bundles, some of the brown content (tannin) from ground tissue, crystal fibers and lignified fibers.

ACKNOWLEDGEMENT The authors are thankful to Director, IPGT&RA, Gujarat Ayurved University, Jamnagar and Department of AYUSH, for providing financial support and other facilities to carry out the research work. We express our thankfulness to Mr. B. N. Hota, Rtd. DFO, Govt. of Odisha; Mr. Pareswar Sahoo Pharmacognosy expert; Mr. Malaya Das, Forest Range Officer, Govt. of Odisha and other traditional healer who helped us during drug collection at Gandhamardan Hills, Balangir and Bargarh, Odisha.

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REFERENCES Anonymous, (1990), Glimpses of medicobotany of Bastar district, Madhya Pradesh, CCRAS publication, pp. 114.

Saxena H.O, (1995), The Flora of Orissa, Regional Research Laboratory, Bhubaneshwar, 1st edition, pp. 288.

Anonymous, (1999), The Ayurvdic Pharmacopoeia of India, Govt. of India publication New Delhi, 1st edition, Vol. I, Appendix 2.

Trease GE, Evans WC (2002), Trease and Evans Pharmacognosy, Harcourt brace & Co. 99, 512, 547.

Asia, Pvt. Ltd. and W.B. Saunders Company Ltd., Edition 15, pp. 32, 33, 95–Johnson Alexander Donald, (1940), Plant Micro technique. New York, London, Maccgrow Hill Book Company, pp. 105. Khandelwal K.R., Kokate C.K., Gokhale S.B. (1996). Practical pharmacognosy techniques and experiments. Nirali Prakashan, Pune, pp. 10–39. Kirtikar & Basu, (2003), Indian Medicinal Plants, Bishen Singh Mahendra Pal Singh, Dehra Dun Vol. I, pp 568.

Tribhubana Panda, Rabindra N Pandhy, (2007), Sustainable food habits of the hill dwelling kandha tribe in kalanandi district of Orissa, Indian journal of traditional maedicine, vol. 6(1), pp. 103–105. Veeramuthu Duraipandiyan, Muniappan Ayyanar, Savarimuthu Ignacimuthu, (2006), Antimicrobial activity of some ethnomedicinal plants used by Paliyar tribe from Tamil Nadu, India, BMC Complementary and Alternative Medicine, 6:35 doi:10.1186/1472-68826-35 Wallis T E. (1985), Textbook of Pharmacognosy, London Churchill Publication, pp.572–82

Source of Support: Department of AYUSH,

Conflict of Interest: None Declared

Govt. of India

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Research article A CLINICAL EVALUATION ON RUJAKARA MARMA WITH SPECIAL REFERENCE TO PAIN THRESHOLD Benjwal Shobha1* 1

Asst. Prof. Rachana Sharir Dept., M.S.M. Institute of Ayurveda, Khanpur Kalan, Sonipat Haryana *Corresponding Author: E-mail: shobha.benijwal@yahoo.in

Received: 10/03/2013; Revised: 18/02/2013; Accepted: 20/03/2013

ABSTRACT Marmas are vulnerable spots, constituting the essential aspect of surgico-anatomical knowledge. While defining the type of marmas based on the effect or prognosis of the trauma, Acharya Sushruta has classified five types of marmas. Among them he has appreciated pain as a residual effect of trauma in Rujakara marma. This study was planned to analyze the tissues which were responsible for pain in Rujakara marma & to evaluate pain-threshold relation with this marma. The study involved 60 healthy individuals who volunteered. According to anatomical sites, in Group I was taken to evaluate pain threshold at 8 (Eight) Rujakara marmas (vital spots which have pain due to trauma) sites of body and In Group II the same 60 individuals were taken to measure pain threshold at sites other than Rujakara marmas. The results have shown that eight sites of Rujakara marma has different types of fibrous Scleratogenic tissues having high pain threshold noceceptive impulses, which might be the probable reason for Rujakara marmas to have a high pain-threshold in comparison to other sites of the body. KEYWORDS: Ruja, Painthreshold, noceceptive impulse

Cite this article: Benjwal S (2013), A CLINICAL EVALUATION ON RUJAKARA MARMA WITH SPECIAL REFERENCE TO PAIN THRESHOLD, Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 254â&#x20AC;&#x201C;258

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 3 | March 2013 | 254–258

INTRODUCTION The Ayurvedic Science of Marma is itself a treatise on Surgico-anatomical learning. The concept of marma is a great contribution of Sushruta, who mentioned 107 vital points in various parts of the body, which should be carefully dealt with during surgery & should always be protected from injury, as the essence of life (prana) rest in them (Sushruta 200 B.C). Though general definition of Marma signifies that every marma is the confluence of five types of tissue, namely mamsa (muscle) Sira (vessels), Snayu (ligaments) Asthi (bone) & Sandhi (joints) (Charaka 200 B.C); but it is evident from the description of injuries that the traumatic effect or prognosis entirely depends on the predominance of the tissue type at the marma. Sushruta has classified Parinam prakar marmas (residual effect of trauma) into sadhyo pranhara (death on the spot after trauma) Kalantara pranhara,(death occuring after a short period post-trauma) Vishalyghana (death occuring after removal of foreign body from a traumatic wound) Vaikalyakara (there will be a stable deformity in the body structure post-trauma) & Rujakara. The vitiation of Rujakara marma creates only the feeling of pain and there is no condition of death or morbidity (Sushruta 200 B.C). There are eight points of Rujakara marma in the body out of which Gulfa (Ankle joint) and Manibandh (wrist joint) are sandhi & Kurchshira (brush like structure) are of Snayu predominance. The Rujakara marmas possess properties of Agni S.No

Inclusion Criteria

(Fire) & Vayu (Air) Mahabhutas both of which causes pain as a residual effect of trauma (Sushruta 200 B.C). The perception of pain due to trauma depends upon many factors like pain receptors in the skin (mechanical, Chemical & Thermal receptors) & rate of tissue damage (Snell S.Richard, 1992). The differentiation without discrimination was assessed on the basis of gradation of pain-threshold & in the present work this criterion was adopted for differentiating tissues at different anatomical sites to understand ‘Rujakara marma’ in a better way. Hence a study was planned to differentiate out the tissues responsible for Rujakara marma and to study the pain threshold at Rujakara marma sites & compare it with pain threshold in other body parts. MATERIALS The research work was conducted on 60 healthy individuals selected from Indira Gandhi Girls Hostel, P.G.Hostel of State Ayurvedic College, Lucknow. Some of the observations have been made by the researcher itself at the neighbourhood. All the 60 healthy individuals were taken in two groups: Group I-Measurement of Pain threshold at site of Rujakara marma sthana Group II- Measurement of Pain threashold 5 cms proximal to the site of Rujakara marma sthana

Exclusion Criteria

Individual between age of 18 – 40 years Individual below 18 or above 40 years.

Either Sex

Pregnant women

Having no disease

Any medical surgical co-morbidity

For this study a Proforma was prepared. The information regarding name, age, sex and occupation were noted down. The blood

If taking any medicine

pressure, temperature, respiratory rate & pulse rate were also recorded. For the purpose of measuring pain threshold following instruments

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 3 | March 2013 | 254–258

were used: Sphygmomanometer, Specially designed blunt conical wooden object, Scale, Stethoscope, Watch. (River.J, 1999). METHOD Measuring of Pain Threshold: The subjects were asked to sit on a chair in erect posture placing forearm in supinated position on a table. A specially designed blunt conical wooden object with cuff of sphygmomanometer was kept on the site of Rujakara marma i.e. at the wrist joint (for manibandh marma) & just medial to the tubercle of scaphoid at the palmar surface (for kurchashira marma) (Solanki J.C.1982). Thereafter, cuff was wrapped & air was gradually pumped to produce, pressure pain. As soon as the subject complained the pain, the

pressure necessary to produce that degree of pain was recorded in terms of mm Hg. These readings were utilized as the parameter of painthreshold. For the measurement of pain threshold in lower extremities the wooden conical object was placed on Ankle joint (For Gulfa marma) & at the cross of line drawn horizontally on the planter surface by joining the medial conversity of medial cuniform bone and base of the 5th metatarsal bone and vertically following the junction of 1st and 2nd toe (For Kurcha Shira marma) (Solanki J.C.1982). For accuracy three readings were taken at the same place at a 10 min interval. For comparative studies, measurement of pain threshold was done on other body parts i.e. 5 cm proximal to Rujakara marma site. Average of all these readings of pain-threshold and observations were statistically calculated.

Table-1 Statistical analysis of Pain Threshold in Group I and Group II S.n o

Group I

Group II

‘t’ value

‘p’ value

Name of Mean value S.D Rujakara of pain marma threshold

5cm proximal Mean S.D to Rujakara value of marma pain threshold

Manibandh (Wrist joint)

132.02

± 46.23

Manibandh (Wrist joint)

128.99

± 48.58

18.52

<0.001

Gulfa Marma 151.71 (Ankle Joint)

± 52.07

Gulfa Marma 118.25 (Ankle Joint)

± 49.60

13.30

<0.001

Kurcha shira 130.27 (Upper extremity)

± 48.64

Kurcha shira 91.78 (Upper extremity)

± 45.68

7.14

<0.001

Kurcha shira 170.65 (lower extremity)

± 68.88

Kurcha shira 120.95 (lower extremity)

± 60.18

9.68

<0.001

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 3 | March 2013 | 254–258

Table-2 Comparative Assessment of Pain threshold in Group I and Group II S.No. Name of the Site of Pain Threshold Mean value of Pain Threshold S.D 1.

Rujakara marma

136

± 6.23

Control site of Rujakara marma

128.99

± 48.85

t value = 4.237 p value = <0.001

RESULTS Table-1 indicates that the average pain threshold at Manibandha (wrist joint) was 190.36 mm Hg with a SD of ± 56.40 and 124.22 mm Hg with a SD of ± 46.60 was found at a site, 5 cm proximal from Manibandha Marma (wrist joint). Comparative assessment was statistically significant with ‘t’ value of 18.52 and p value < 0.001. The average of pain threshold at Gulfa (ankle joint) was 151.71 mm Hg with a S.D. of ± 52.07 and for sites other than Gulfa (ankle joint) i.e., 5 cm proximal to the Gulfa Marma (ankle joint) was lower with a mean value 118.25 mm Hg with a S.D of ± 49.60. The difference observed was highly significant, with ‘t’ value being 13.30 and p value <0.001. The mean value of pain threshold was 130.27 mm Hg with a S.D. of ± 48.48 and for site other than Kurchshira Marma of Upper extremity while it was 91.78 mm Hg with a S.D. of ± 45.68 for the other sites on the palm i.e. 5 cm distal from the Kurchashira Marma of Upper extremity. The comparative assessment shows the ‘t’ value being – 7.149 and p value < 0.001, which is highly significant. The mean value of pain threshold at Kurchshira Marma of lower extremity was 170.65 mm Hg with a S.D. of ± 68.88 and other site on planter surface 5 cm distol to Kurchshira Marma of lower extremity while it was 120.95 mm Hg with a S.D. of ± 60.18. The comparative assessment shows the value were statistical significant with a ‘t’ value of 9.685 and p value < 0.001. In Table-2 the mean value of pain threshold at Rujakara marma site was

136.02 mm Hg with S.D. of ± 46.23. The mean value of pain threshold of control group of Rujakara marma was 128.88 mm Hg with S.D of ± 48.85. So it was observed that the average pain – threshold of control site of Rujakara marma was lower than the average pain threshold at Rujakara marma sites. The comparison between the two groups is highly significant with a ‘t’ value of 4.237 and p value < 0.001. The mean value of pain threshold at Rujakara marma site was 136.02 mm Hg with S.D. of ± 46.23. The mean value of pain threshold of control group of Rujakara marma was 128.88 mm Hg with S.D ± 48.85. So it was observed that the average pain – threshold of control site of Rujakara marma was lower than the average pain threshold at Rujakara marma sites. The comparison between the two groups is highly significant with a ‘t’ value of 4.237 and p value <0.001. DISCUSSION Based on the result of vitiation of marma sites, the marmas are divided into 5 types in which the Rujakara marma belongs to least morbidity (Sushruta, 200 B.C). Ruja (pain) is a expression of body related to damage of tissue. This is a psychosomatic phenomenon which is different in every individual (www.iasppain.org), which has been made the criteria in this study to differentiate one site of tissue to another site of tissue. This phenomenon has been used as a basic tool by school of Sushruta to classify Marmas (vital spot) on the basis of result of the traumatic effect. All the tissues

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such as Mansa (Muscle) Sira (vessels) Snayu (ligament) Asthi (bone) Sandhi (joints) carry noceceptor a biological sensor which is related to noxious stimuli which is cast by mechanical (direct trauma) biological irritation & thermal The nonceceptive receptors are specialized nerve endings in skin and deep tissues and unlike other sensory receptors they are activated at high threshold by a range of potentially damaging stimuli (Grey’s, 1980). This reveals that school of sushruta has concerned nonceceptive pain as a criterion for Rajukar marma. There are eight Rajukar marmas being comparatively high nonceceptive pain-threshold which also indicates that it belongs to sclerotogenous pain rather dermatogenic pain. The sclerotogenic pain pattern is selective of ligament, tendon, disc, periosteum & apophysial joint (Grey’s 1980) the anatomical places of Rujakara marma carry one or more than one type of sclerotogenous tissue like manibandh (wrist joint) and gulfa (ankle joint) carry abundant ligaments and kurch shira consists largely tendons and all these have sclerogenatic pain

receptors having high pain threshold. (Snell S Richard 1992). This discussion lead to Sushruta’s observation in the form of Rujakara marma that the vital parts are precisely of sclerotogenous type of nonceceptive pain receptor, bearing high threshold as compared to other places where tendon, ligament & other fibrous structures are lesser in quantum. The Group II are largely consisting of muscle & vascular tissue which are definitely having lesser pain-threshold as compared to Group I. CONCLUSION The classified Rujakara marmas according to school of Sushruta anatomically placed at eight sites carrying different fibrous sclerotogenic tissues, have comparatively high threshold of noceceptive impulse. This observation further draws the attention to the subject of inquiry that whyManibandha (wrist) has the highest pain threshold than Kurchashira of upper extremity. This study opens the gate of further research to prove these specific observations made in present study.

REFERENCES: (200B.C) Chakrapani Tika commentary by Tripathy Brahmanand (2002) on Charak Samhita 6th edition, published by Chaukhambha Subharti Prakashan 1999, Varanasi, Vol. II Chikitsasthan Chapter-26th page 720– 734.

Solanki J.C, A Study on Rujakara marma with Special Reference ot their anatomical & surgical significance. Thesis M.D Lucknow University, 1982.

Grey Henery (1980) Grey’s Anatomy by Peter L Williams and Roger Warwick 36th edition, Churchill Livingstone, Published by Jarrold and Sons Ltd.

Sushruta (200 B.C.) Sushruta Samhita hindi commentary by Ghanekar B.G.(1999), 14th edition, published by Mehar Chand Luxmi Chand Publication, Chapter-6th page-184–189.

Charak

Snell.S.Richard (1992) Clinical Anatomy for Medical student 4th edition Chapter-9, page- 487–495. Source of Support: Nil

Source: //www..iasp-pain.org/. Basic concept of Pain physiology.

River J (1999) Adolescence pain measurement, pain threshold. Journal of paediatrics 75(4):244–248 Conflict of Interest: None Declared

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Research article PHARMACOGNOSTICAL AND PRELIMINARY PHYTOCHEMICAL INVESTIGATIONS ON DIFFERENT PARTS OF BULBOPHYLLUM NEILGHERRENSE WIGHT. -AN ORCHID USED IN FOLK MEDICINE. Kumari Harshitha1*, Nishteswar K2, Harisha C R3 1

Ph D Scholar, Department of Dravyaguna, IPGT&RA, GAU, Jamnagar.Gujarat, India Professor and HOD, Department of Dravyaguna, IPGT&RA, GAU, Jamnagar. Gujarat, India 3 Head, Pharmacognosy, IPGT&RA, GAU, Jamnagar. Gujarat, India *Corresponding author: drharshitharai@gmail.com, Ph: 8460832302. 2

Received: 11/02/2013; Revised: 22/02/2013; Accepted: 28/03/2013

ABSTRACT Bulbophyllum neilgherrense Wight. is an epiphyte belonging to the family Orchidaceae found growing on medium to large sized host trees endemic to the forests of Western ghats which is yet to be scientifically explored. A few published reports are available with regards to its medicinal uses and scientific studies. In the present study, systematic pharmacognostic evaluation of leaf, stem and root of the plant were carried out with respect to macroscopy, microscopy and preliminary phytochemical screening. Macroscopic study detailed the structure of leaf, stem and root. Microscopic study demonstrated the presence of mucilage cells and calcium oxalate crystals in leaf; oil globules, tannin and mucilage cells in stem; velamen tissue, passage cells and lignified parenchyma cells in root. Qualitative phytochemical investigations showed the presence of alkaloids, tannin and phenol in methanol extract of stem and root whereas in leaf, constituents like alkaloids, saponin glycosides, tannin, phenols and reducing sugar were observed. The morphological, histological and phytochemical investigations reported in the paper may become supportive to establish the authenticity of the plant simultaneously giving a wide scope for further pharmacological and clinical researches. KEY WORDS: Bulbophyllum neilgherrense, leaf, stem, root, pharmacognosy, phytochemical studies.

Cite this article: Kumari H, Nishteswar K, Harisha C R (2013), PHARMACOGNOSTICAL AND PRELIMINARY PHYTOCHEMICAL INVESTIGATIONS ON DIFFERENT PARTS OF BULBOPHYLLUM NEILGHERRENSE WIGHT. - AN ORCHID USED IN FOLK MEDICINE., Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 259–269

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INTRODUCTION Bulbophyllum neilgherrens Wight. belonging to Orchidaceae family, commonly known as Pottlekai in Kannada (Bhat Gopalkrishna, 2003), Kalmel pullurvi in Malayalam is abundantly available in Western ghats (Rajendran A et al., 1997) and sparsely distributed in Eastern ghats (Jadhav S N, 2003). Forests of Karnataka, mainly Udupi (Bhat Gopalkrishna, 2003), Belgaum (Gamble J.S, 2011), Malabar (Hooker J D, 1885) are the places from where the species can be traced. The plant is endemic to South India, which occurs in plains and in higher elevations up to 900 m (Abraham and Vatsala, 1981). The special characteristic of the plant is the presence of pseudobulb for the preservation of water and nutrients. Pseudobulbs are 3–3.5 cm long and 2 cm across, smooth, green, four angled. Progressing yellowing of pseudobulbs is observed on ageing. Leaves 10–15cm long, 2–3 cm broad, coriaceous, elliptic to broadly oblong, obtuse at apex, base narrowed tapering into short petiole attached to the pseudobulb (Bhat Gopalkrishna, 2003, Abraham and Vatsala, 1981). Scape stout, from the base of the pseudobulb, sheathed at the base, jointed and with bract-like sheaths at the joints. Flowers in racemes, petals small, pale yellow, lip purple (Theodore cooke, 2006). Karyomorphological study of this orchid species has shown the chromosomal number to be 19 (Abraham and Vatsala, 1981). The plant is used by the folk people for restoration of adolescence and as tonic in the form of juice extracted from pseudobulb (Hossain MM, 2011). The paste prepared from pseudobulb and leaf is consumed along with cow’s milk to treat leucoderma (Rajendran A et al., 1997). In certain regions of Karnataka, various parts of this orchid are used by the villagers in the management of heart diseases (Kumari Harshitha, 2011). Scarce information is available regarding the phytochemical and pharmacological profiles of this drug. The methanolic extract of leaf showed the presence of flavonoids and

cyanogenic glycosides (Maridass M et al., 2008). In vitro study of ethanol extract of leaf and pseudobulb showed pronounced antibacterial effect as compared to the effect produced by chloroform and aqueous extracts (Priya K et al., 2005). No information is available regarding the microscopical characters of the plant. In this regard, pharmacognosy including transverse section, powder microscopy, histochemical tests of leaf, stem and roots with their preliminary phytochemical investigations were carried out with a view to establish purity and standard of the sample.

MATERIAL AND METHODS Plant material Whole plant was collected from its natural habitat in Puttur TQ, Karnataka, India during the month of April 2012. The botanical identity was confirmed by the botanist Dr. K. Gopalakrishna Bhat, Professor of Botany (Rtd.), Poorna prajna college, Udupi, India. A herbarium specimen was preserved in the Pharmacognosy lab, IPGT&RA, Gujarat Ayurved University, Jamnagar with voucher specimen number 6025\2012. Macroscopical evaluation (Kokate et al. 2005): The sample was cleaned and macroscopic evaluation of whole plant was carried out. The leaf, stem and root were then separated and individual macroscopic characters like size, shape, texture were noted in detail. Microscopical evaluation (Trease and Evans 2009, Wallis 1985): Free hand sections of leaf, stem and root were taken and washed with chloral hydrate solution. Sections were first observed in distilled water then stained with phloroglucinol and conc. HCl. Powder microscopy of shadedried powder was also carried out. Photomicrographs were taken by Carl zeiss trinocular microscope.

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Histochemical tests (Krishnamurty 1988): Thick sections were treated with various reagents to locate chemical constituents i.e. Tannin, mucilage, lignin and calcium. Pharmaceutical 2000):

evaluation

(Anonymous,

Physicochemical parameters and preliminary phytochemical investigations were conducted on shade dried powders of leaf, stem and root. The presence or absence of different phyto-constituents in aqueous & methanol extracts was detected.

RESULTS Macroscopical characters- [Fig 1] Bulbophyllum neilgherrens Wight. is an epiphytic rhizomatous orchid with greenish angled pseudobulbs bearing a single leaf at its apex. Roots arise from the base of pseudobulb. Scape is longer than the leaf which emerges from the base of the pseudobulb. It is sheathed at the base and sheath is also seen at the joints. Inflorescence is raceme, drooping with many flowers. Fruit is a capsule, green, globularelongate, 2 cm long, 5 angled with minute seeds. [Fig 1A & 1B]

Description of leaf- Leaves are 8 cm–15 cm long, 2–3 cm broad, coriaceous, elliptic to broadly oblong, flattened, succulent, obtuse at apex with narrow base. Midrib is prominent in the ventral surface where as grooved on the dorsal surface. Leaf margins are simple. Leaf blade has parallel venation, tapers into short petiole attached to the pseudobulb. A single leaf emerges from the top of each pseudobulb. [Fig 1C] Description of stem- Horizontally creeping stout somewhat rhizomatic, measuring about 6– 8 × 0.2–0.3 cm, rounded with slight ridges and grooves, hard, light brown in colour with dark brown scaly rings formed at successive intervals. Nodal region is bulged, flattened gives cone shaped structure inside the bulb. Tuft of roots arise from the lower side of the node. [Fig 1D] Description of root- Tuft of thin roots arise from the nodal region of the stem (adventitious) measuring about 6–10 cm long, greenish to light brown in colour, covered with delicate fibrous type of absorptive tissue called velamen, which is dead and perforated. When the root becomes dry velamen part detaches from the thin wiry central part which is strongly attached to the stem. [Fig 1E]

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Microscopical characters-

Stem [Fig 3]

Leaf [Fig 2]

TS of stem showed the outer epidermis is followed by hypodermis and the ground tissue [Fig 3A]. Epidermis is made of single layered compactly arranged barrel shaped cells without intercellular spaces and is covered with thick cuticle. Hypodermis made of 6–7 layers of compactly arranged lignified sclerenchyma cells seen below epidermis [Fig 3B]. Endodermis is made of abruptly placed barrel to uneven shaped cells with thick lignified cells followed by ground tissue. Ground tissue is consisting of outer thin walled parenchymatous zone, then pericyclic fibre zone and the central region with scattered vascular bundles. Ground tissue occupied 2/3rd portion of the section lying beneath the endodermis. 5–7 layers of loosely arranged parenchyma cells without intercellular spaces are seen consisting oil globules, tannin [Fig 3C]. Some of the inner parenchyma cells were lignified and pitted. 2–3 layers of lignified pericyclic fibres forming a ring like structure were noticed. Number of vascular bundles are scattered all over the zone. Vascular bundles in peripheral zone are larger in size than those in the central zone. Vascular bundles are collateral and closed, each bundle surrounded by a sheath which is more conspicuous towards upper and lower side of the bundle. The Vascular bundle consists of xylem and phloem. Xylem consisting 2–3 large metaxylem and small protoxylem with tracheids and lysogenous cavity is present inner to the protoxylem. Phloem consists of conspicuous sieve tubes and companion cells. Phloem parenchyma is not found. Mostly the ground tissue is parenchymatous; some of the lignified pitted parenchyma surrounds the vascular bundles. Rest of the parenchyma cells were also lignified and are thick walled [Fig 3D].

Leaf was isobilateral in nature. The upper and lower epidermis was undifferentiated, mesophyll tissue is filled with chloroplast pigments consisting number of secretory cells. Vascular bundles were centrally located. Upper epidermis was single layered with compactly arranged barrel shaped cells. At the midrib portion the barrel shaped cells were morphologically modified into tangentially elongated compactly arranged cells resulting in the formation of motor cells or hinge cells. These epidermal cells were covered with thick, ridged cuticle. Lower epidermis has only the barrel shaped cells without motor cells, with thick cuticle [Fig 2A]. Both the epidermis possesses sunken stomata. Mesophyll was spirally thickened near the epidermis with banded parenchyma cells. Rest of the mesophyll tissue was filled with undifferentiated isodiametric parenchyma cells containing numerous choloroplast, and was compactly arranged. The mesophyll also consists of some mucilage cavities. The mesophyll also consists of raphide idioblast. In midrib portion there was a centrally located large vascular bundle and smaller vascular bundles are passing through the main nerves [Fig 2B]. Vascular bundle consists of phloem towards lower epidermis, xylem towards upper epidermis with few xylem elements with xylem parenchyma and its fibres, where as phloem with few sieve elements and fibres [Fig 2C]. Some of the mesophyll parenchyma consists of reddish brown colour contents. The thick cuticle and sunken stomata shows the xerophytic nature of the plant, where as the parenchyma cells, large mucilage cells show the hydrophytic nature of the plant. Powder microscopy: Diagnostic characters of powder showed anisocytic stomata in epidermis [Fig 2D], fibers from vascular bundles [Fig 2E], acicular crystals of calcium oxalate [Fig 2F], raphides, mucilage cells [Fig 2G], annular thickened parenchyma cells, tannin from mesophyll tissue and epidermal cells in surface view.

Powder microscopy: Powder showed the presence of acicular crystals, tannin, fragments of annular vessel, tracheids with oil globules [Fig 3E], fibers [Fig 3F], fragments of parenchyma cells, annular pitted vessel[Fig 3G] and lignified parenchyma cells [Fig 3H].

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 259â&#x20AC;&#x201C;269

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 259â&#x20AC;&#x201C;269

Root [Fig 4] TS of root showed outer velamen followed by single layer of exodermis, leading into a wide zone of cortex. Inner to it there was circularly arranged single layer of endodermis followed by pericycle and the vascular bundles

with alternatively arranged xylem and phloem, forming central large pith [Fig 4A]. The outer layer of the root showed velamen tissue. These are dead cells, variously elongated, thick walled and compactly arranged. Exodermis is the outermost layer of the cortex, tangentially

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arranged, thick walled and suberized. Few cells which are unthickened are passage cells. Beneath the exodermis many layers of (5–6) loosely arranged parenchyma cells, consisting chloroplast pigments, along with some air chambers are present. Some of the parenchyma cells have raphides, oil globules, starch grains or yellowish brown tannin material [Fig 4B, Fig 4C]. Endodermis is single layered with compactly and circularly arranged barrel shaped cells forming a ring, followed by single layer of thin walled pericycle. Some of the endodermal cells opposite to phloem are thickened. Vascular bundles form polyarch, radially arranged and exarch. Xylem vessels with metaxylem towards pith and protoxylem towards the endodermis. Xylem alternate with the phloem, xylem consists of xylem parenchyma and tracheids. The xylem parenchyma is angular. Phloem consists of sieve tube and companion cells. Pith is the

central most part of the root being angular parenchymatous, thick walled and lignified [Fig 4D]. Powder microscopy: Diagnostic characters of powder showed the presence of lignified fiber [Fig 4E], oil globules [Fig 4F], tannin and starch grains [Fig 4G], acicular crystals, tracheids [Fig 4H], mucilage containing cell, fragments of pitted vessel [Fig 4I]. Histochemical testsThe results of various histochemical tests conducted on the leaf, stem and root powder are depicted in Table I. Pharmaceutical evaluation The physicochemical parameters and qualitative analysis results are enumerated in Table II and Table III respectively.

Table: I showing Histochemical test results Sl. no

Reagent

Observation

Characteristics

Result Leaf Stem Root

Red

Lignified cells

Blue Dissolved

Starch grains Calcium oxalate crystals Tannin cells

− ++

++ ++

Phloroglucinol+Conc. HCl Iodine Phloroglucinol+Conc. HCl Fecl3 solution

Ruthenium red

Mucilage

2. 3.

Dark blue to black Red

Table: II showing results of physicochemical parameters Sl. no 1 2 3 4 5 6 7

Parameters Foreign Matter Loss on Drying % w/w Total Ash Content% w/w Acid Insoluble Ash % w/w Water Soluble Extractive Value % w/w Alcohol Soluble Extractive Value % w/w PH

Leaf Nil 5.844 6.05 0.1 33 5.9 5

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Stem Nil 10.009 0.499 0.0998 3.7 6.8 5

Root Nil 5.638 2.497 0.4995 4.7 3.7 5

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Table : III showing results of phytochemical evaluation Sl. no Parameters Leaf Stem Alkaloids (M.E.)* + + 1 Alkaloids (W.E.)* − − 2 Saponin Glycosides (W.E.) + − 3 Tannins &Phenols (M.E.) + + 4 Tannins &Phenols (W.E.) + − 5 Flavanoid (M.E.) − − 6 Steroids (M.E.) − − 7 Reducing sugar(W.E.) + − 8 * M.E- Methanol extract; W.E- Water extract

DISCUSSION B. neilgherrens Wight. is an epiphytic orchid with stout rhizome modified into pseudobulb to store moisture in excess so that it can survive in unfavorable seasons. The plant usually found growing in humid conditions of moist deciduous and evergreen forests on tree trunks of Anacardium occidentale (Cashew), Syzigium cumini (Jamun) or other medium sized to large trees. A very few indigenous medicinal claims have been recorded on leaves and pseudobulb of the plant. Limited number of scientific studies has been reported on this species with regard to structural, chemical, pharmacological or clinical evaluation. Pharmacognostical study reveals that the leaf is isobilateral. Epidermis has some motor cells and the mesophyll is undifferentiated with spirally thickened parenchyma cells, which shows that the cells are able to retain water for long duration. The motor cells help the leaf to roll due to the changes in their turgidity there by reducing the stomatal transpiration under xeric conditions. Presence of mucilage cells, calcium oxalate crystals, sunken stomata and the absence of trichomes are the important characters of leaf. Oil globules and tannin containing cells, calcium oxalate crystals, mucilage containing cells are the important characters found in rhizomatic stem. Velamen tissues in roots during dry weather remains filled with air and during rain quickly absorb water. Passage cells serve as channels for flow of water absorbed by the velamen. Lignified

Root + − − + − − − −

parenchyma cells in the pith region of the root are the other special characters. These may help the species to adapt itself in the stress of climatic variations. Microscopic study of the plant powder shows the presence of large quantities of fibers, oil globules, tannin, and mucilage containing cells. Histochemical tests of leaf, stem and root shows the presence of lignin, tannin, calcium and mucilage. Starch was detected only in root. The physicochemical parameters shows that loss on drying is more in stem as compared to leaf and root which shows that higher moisture content is in stem followed by leaf. Total ash is more in leaf followed by root and less in stem. Water soluble extractive value is more than five times higher than alcohol soluble extractive value in leaf showing the presence of more water soluble constituents. Root also shows higher extractive value in water where as stem has higher extractive value in alcohol. PH value of leaf, stem and root did not show any difference. Among the qualitative assessment, alkaloids were detected in methanolic extracts of all three parts of the plant taken for investigation, where as it could not be detected in water extract. Stem and root shows the presence of tannin and phenols only in methanol extract while leaf contains tannin and phenols in both water and methanol extract. Water soluble extractives being highest in leaf, also shows the presence of saponin glycosides and reducing sugar in water extract.

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Natural products with reservoirs of structural and chemical entities will have definite therapeutic relevance. Tannins are reported as potential antiviral, antibacterial, antiparasitic and hypolipidemic (Tannin.2013), Pengelly Andrew 2004). Tannin and saponin containing drugs demonstrated anti diabetic activity (Akhlaghi Farideh et al., 2012, Pengelly Andrew 2004). Saponins, tannin and phenols exhibit antibacterial activity (Doughari JH et al., 2007). Research studies also supported the view that phyto-constituents like phenols, saponins, tannins, alkaloids exhibit antioxidant, adaptogenic and antimicrobial activities (Sukh Dev, 2006).

CONCLUSION The studies carried out on the sample not only established the appropriate data that may be utilized for identification, but also established the purity and standard of the plant sample. Based on the reported phytoconstituents some more pharmacological as well as clinical studies may be carried out for producing a proper scientific validation of the folk orchid Bulbophyllum neilgherrense wight. ACKNOWLEDGEMENT The authors acknowledge Dr. K. Gopalakrishna Bhat, Professor of Botany (Rtd.) for his help in identification of this orchid species and its authentication, Villagers of Sullia TQ for sharing their knowledge and experience on the plant.

REFERENCES Abraham and Vatsala. (1981), Introduction to Orchids. Tropical Botanic Garden and Research Institute, Pp 533. Akhlaghi Farideh, Rajaei Ziba, Mousa-Al-Reza Hadjzadeh, Mehrdad Iranshahi and Mahadi Elizabeth. (2012), Antihyperglycemic effect of Asafoetida (Ferula assafoetida Oleo gum resin) in streptozotocin- induced diabetic rats, World Applied Sciences Journal, 17 (2): 157–162. Anonymous. (2000), Protocol for testing of Ayurveda, Siddha and Unani medicines.Ghaziabad: pharmacopoeial laboratory for Indian medicines, Department of Ayush, ministry of health and family welfare, Government of India. Bhat Gopalakrishna K. (2003), Flora of Udupi. Indian naturalist, Pp 913.

Doughari JH, Pukuma MS, De N. (2007), Antibacterial effects of Balanites aegyptica L. Drel. and Moringa oleifera Lam. on salmonella typhi, Afr J Biotechnol;6:2212–5. Gamble J.S. (2011), Flora of the presidency of Madras, Shiva offset press, Dehradun. Pp 2017. Hooker J D (1885), The Flora of British India, L. Reeve & Co., Ltd.D-2, vol5, Pp 910. Hossain M M. (2011), Therapeutic orchids: traditional uses and recent advances — An overview, Fitoterapia (82) pp 104. Jadhav

S N. (2003), Medicinal plants conservation and sustainable utilization project, EPTRI-ENVIS news letter, vol.9 No.2.

Kokate C K, Purohit AP, Gokhale SB (2008) Pharmacognosy (42nd Edn), Nirali Prakashan, Pune, pp 99.

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Krishnamurty K.V. (1988), Methods in the plant histochemistry. Vishwanadhan Pvt Limited, Madras, Pp 1–77. Kumari Harshitha. (2011), personal communication with villagers of Sullia TQ, Karnataka, India. Maridass M, Zahir Hussain M I, Raju (2008), Phytochemical survey orchids in the Thirunelveli hills South India, Ethnobotanical leaflets 705–12.

orchids of Southern India. Ancient Science of Life, Vol. No. 17 (1). Sukh Dev. (2006), A selection of prime Ayurvedic plant drugs Ancient-modern concordance. Anamaya Publishers, Pp 501.

G. of of 12:

Tannin. (2013). Wikipedia, the free encyclopedia.Retrieved February, 2013, from http://en.wikipedia.org/wiki/Tannin.

Pengelly Andrew. (2004), Constituents of Medicinal Plants. CABI publishing, Pp 31.

Theodore cooke. (2006), Flora of the presidency of Bombay, Bishen singh Mahendrapal Singh, Dehradun, Vol 2, Pp 1083.

Priya

Trease

G. E and Evans W.C. (2009), Pharmacognosy, 16th Ed. Saunders, Elsevier. Pp 537–562.

Wallis

T.E. (1985), Text book of Pharmacognosy, 5th Ed, CBS Publishers, New Delhi, 1985. Pp.571– 578.

K and Krishnaveni C. (2005), Antibacterial effect of Bulbophyllum neilgherrense Wt. (Orchidaceae). An invitro study. Ancient science of life, Vol XXV (2).

Rajendran A, Rama Rao N, Ravikumar K and Henry A N. (1997), Some medicinal

Source of Support: Nil

Conflict of Interest: None Declared

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Research article A COMPARATIVE ACUTE TOXICITY EVALUATION OF ASHOKA KSHEERAPAKA PREPARED FROM TWO DIFFERENT SPECIES OF SARACA (S. ASOCA & S. THAIPINGENSIS) Chavan Sulakshan S1*, Gamit R V2, Ashok B K3, Shukla V J4, Das P5, Ravishankar B6 1

Ph D Scholar in Ayurvedic Pharmacology, IPGT&RA, Gujarat Ayurveda University, Jamnagar, Gujarat, India 2 Laboratory Assistant, IPGT&RA, Gujarat Ayurveda University, Jamnagar.Gujarat, India 3 Drug discovery lab, R & D, Himalaya drug company, Bangalore, Karnataka, India 4 Head, Pharmaceutical Chemistry Lab, IPGT & RA, Gujarat Ayurveda University, Jamnagar, Gujarat, India 5 Chairman, The Science Foundation For Tribal & Rural Resource Development, Bhubaneswar, Odisha, India 6 Director, SDM Research Centre for Ayurveda and Allied Sciences, Kuthpady, Udupi, Karnataka, India *Corresponding Author: E-mail: sulakshan_2004@yahoo.co.in

Received: 10/03/2013; Revised: 25/03/2013; Accepted: 30/03/ 2013

ABSTRACT Ashoka (Saraca asoca) is an important Ayurvedic drug for treating gynecological disorders. Hence it is economically important. There were reports that it has become quite scarce in several localities and reported to be threatened in North Eastern Region of India. Ashoka bark widely adulterated with other barks & or from same genus of different species. Ashoka Ksheerapaka is one among clinical formulations of the plant Ashoka. Literature review revealed that no toxicity studies have been undertaken on this formulation especially on Ashoka Ksheerapaka made from S. asoca & S. thaipingensis. Because of this the present study was designed to evaluate Ashoka Ksheerapaka made from two species for acute toxicity in Wistar strain albino rats as per OECD (Organization for Economic Co-operation and Development) guideline 425 with 2000 mg/kg as limit test. On 1st day test formulations were administered & observed for any toxicity changes for next 14 days. On 15th day serum biochemical and hematological parameters were estimated. In all the three groups normal weight gain was observed. Both the formulations did not produce any mortality up to the dose of 2000 mg/kg on oral administration. S. asoca increased the Monocyte percentage & Blood Sugar Level (BSL) significantly, while in S. thaipingensis significant increase in Monocyte percentage & significant decrease in the Platelet count in comparison to Normal Control group.The significant increase in Monocyte percentage is in accordance with the property of estrogens to mediate its effect through estrogen receptor in monocytes. KEY WORDS: Ashoka, Ashoka Ksheerapaka, Acute toxicity, Saraca asoca, Saraca thaipingensis

Cite this article: Chavan Sulakshan S, Gamit R V, Ashok B K, Shukla V J, Das P, Ravishankar B (2013), A COMPARATIVE ACUTE TOXICITY EVALUATION OF ASHOKA KSHEERAPAKA PREPARED FROM TWO DIFFERENT SPECIES OF SARACA (S. ASOCA & S. THAIPINGENSIS), Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 270â&#x20AC;&#x201C;277 Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

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INTRODUCTION Ashoka [Saraca asoca (Roxb.) Willd.] is one of the most important Ayurvedic drug for the treatment of various feminine disorders especially in menorrhagia (Kirtikar & Basu, 2001). The word Ashoka means “without sorrow”, a reference to reputation of it‟s bark for keeping a woman healthy and youthful (Shashikant Patwardhan, 2013). The natives and traditional healers of Chhattisgarh use SitaAshoka (the name given to Saraca asoca) mainly in treatment of gynecological disorders (Nalin, 2005). Its bark is bitter, astringent and sweet in taste. It has stimulating effect on endometrial and the ovarian tissue. It is useful in internal bleeding, hemorrhoids, ulcers, uterine affections, menorrhagia especially due to uterine fibroids, meno-metrorrhagia, leucorrhoea and pimples (Govind Das, 1970). Dried stem bark of Saraca asoca (Roxb.) Willd. is a genuine drug collected from wild or cultivated trees, found in Central and Eastern Himalayas, Western Ghats and Deccan (Anonymous, 1986). Number of studies has shown the adulteration of Ashoka bark with barks of other trees, but less on the trees of the same genus. Ashoka bark is widely adulterated with barks of Polyalthia longifolia, ocassionally bark of Ashoka is mixed with Rohitaka bark (Aphanamixis polystachya (Wall.) R.Parker) and Caesalpinia pulcherrima (L.) Sw (Pradhan P et al., 2009). Raw material from wild is mostly collected by local people. There are high and unintentional chances for a mistaken identity of various other species of the same Genus in the name of Ashoka. The drug from the same genus but of different species is difficult to identify. Such close resemblances of Saraca asoca plant is observed with Saraca thaipingensis. So it is imperative to carry out the toxicological study of these drugs (spp.) before being used in therapeutics. Since ancient times, Ashoka is being used in Ayurvedic preparations but comprehensive data on majority of them is not available. Till date no reports on the toxicological study on Ashoka Ksheerapaka (a medicament prepared with

milk, water & plant drug) made from S. asoca & S. thaipingensis Prain. are available. Hence, this study was designed to evaluate Ashoka Ksheerapaka made from two species of Saraca i.e. S. asoca & S. thaipingensis for acute toxicity in Wistar strain albino rats. MATERIAL AND METHODS Plant material The dried bark of S. asoca and S. thaipengensis were procured from Orissa from authentic source and also correct identification was made in Pharmacognosy laboratory attached to the Institute. The herbarium samples of these two species were deposited in the laboratory (S. asoca- voucher specimen no.6024 & S. thaipingensis- voucher specimen no.6023). Dried barks were coarsely powdered and stored in dry air tight container. Preparation of Ksheerpaka The Ksheerapaka (a medicament prepared with milk, water & plant drug) was prepared according to Acharya Sharangdhara by taking one part drug material, adding cow‟s milk 8 times of bark then adding water 32 times of bark, (i.e. 1:8:32). Ashoka bark powder was weighed. Then, in stainless steel pot weighed coarsely powdered bark of Ashoka was taken as one part, then eight times of the bark cow milk was added & 32 times of bark, water was added in it. Then it was boiled till only milk remained & water was evaporated (Tripathi Brahmanand, 2004). Animals Twenty Wistar strain female albino rats, weighing 160 ± 20 g were taken from the animal house attached to the institute (Institute of PG Teaching and Research in Ayurveda, Jamnagar). They were housed in polypropylene cages with stainless steel cover meshes, at 22 ± 3°C with relative humidity of 50–60 %, on a 12 h natural day and night cycle. They were fed with Amrut brand rat pellet feed supplied by Pranav Agro Industries and with tap water ad libitum. The experiments were carried out in

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accordance with the norms of the Institutional Animal Ethics Committee (IAEC), after obtaining its permission (IAEC -04/09-10/PhD1). Study protocol Acute oral toxicity study for both the samples was carried out as per OECD (Organization for Economic Co-operation and Development) guideline 425 with 2000 mg/kg as limit test. Out of twenty animals 6 animals were allotted to normal control (NC) group. In both the test drug groups (i.e. S. asoca & S. thaipingensis), single animals were dosed in sequence usually at 48 h intervals. Using the default progression factor, doses were selected from the sequence 175, 550, and 2000 mg/kg (because no estimate of the substance‟s lethality was available, dosing was initiated at 175 mg/kg) as recommended in OECD Guidelines 425. Food, but not water was withheld for overnight before the experiment and further 2 h after administration of test drug. As there was no mortality observed even at 2000 mg/kg, additional 4 more animals were dosed with 2000 mg/kg and observed for 14 days with different parameters. The animals were observed continuously for 6 hours after the dosing. The careful cage side observation was done without disturbing the animal attention and at the end of every hour the animals were individually exposed to open arena for recording the behavioural changes. On 14th day evening the rats were kept in metabolic cages for fasting. On 15th day body weight of each animal was recorded. Blood was collected by supra-orbital puncture with the help of micro capillary tubes under mild ether anesthesia for estimation of serum biochemical and hematological parameters. To estimate haematological parameters 0.08 ml blood was mixed with 0.02 ml of EDTA (33.33 mg/ml) and fed to the auto analyzer (Sismes KX-21, Trans Asia). The parameters measured were; Total WBC count, differential leucocyte count, Total RBC count,

haemoglobin content, PCV, MCV, MCH, MCHC and platelet count. For estimation of biochemical parameters, serum was separated from collected blood and requisite quantity of serum was fed to the auto analyzer (Fully automated Biochemical Random Access Analyzer, BS-200; Lilac Medicare Pvt. Ltd., Mumbai) which was automatically drawn in to the instrument for estimating different parameters. Biochemical parameters like blood sugar (BSL) (Pennock CA et al., 1973), serum cholesterol (Roeschlau P et al., 1974), serum triglyceride (Fossati P & Prencipe L, 1982), HDL cholesterol, blood urea, serum creatinine (Slot C, 1965), serum glutamic pyruvic transaminase (SGPT) (Burtis CA & Ashwood ER, 1999), serum glutamic oxaloacetic transaminase (SGOT) (Tietz NW, 1995), serum total protein (Tietz NW, 1986), serum albumin and serum globulin (Doumas BT, 1972), serum alkaline phosphatase (Wilkinson JH, 1969), total billirubin (Pearlman PC & Lee RT, 1974), uric acid (Kabasakalian P, 1973), were estimated. Statistical analysis The results are presented as Mean ± SEM. The generated data were analyzed by employing student‟s t test for unpaired data. One way ANOVA was also employed with Dunnets‟ multiple t test (DMTT) as post-hoc test. For this purpose Sigma-stat software (version 3.1) was employed. RESULTS In all the three groups normal weight gain was observed (Table no. 1). Data pertaining to the effect of test drug on WBC related parameters are given in Table no.2. Both the test drugs increased the Monocyte percentage significantly in comparison to Normal Control group. While a non-significant increase in Total WBC count, Neutrophils percentage & decrease in Lymphocyte percentage, Eosinophils percentage was observed.

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Group NC S. asoca S. thaipingensis

Table - 1 : Effect of test drug on Body weight (BW) Body weight at different time duration Initial (g) On 7th day (g) Final (g) 161.67 ± 5.85 171.00 ± 6.98 173.00 ± 4.31 166.40 ± 8.45 174.40 ± 3.49 188.00 ± 4.20 162.80 ± 7.79 172.40 ± 4.45 180.80 ± 7.97

% change in (BW) 07.01 ↑ 12.98 ↑ 11.06 ↑

(Data: Mean ± SEM, ↑ :- Increase, ↓ :- Decrease)

Table – 2 : Effect of test drug on WBC related parameters WBC related Group Parameter NC S. asoca S. thaipingensis 5660.00 ± 647.77 6880.00 ± 561.61 (21.55↑) 7060.00 ± 553.72 (24.73↑) Total WBC(/cumm) 22.40 ± 3.61 24.60 ± 3.83 (9.82↑) 26.00 ± 3.85 (16.07↑) Neutrophils(%) 71.40 ± 3.25 68.40 ± 3.96 (4.20↓) 67.20 ± 3.74 (5.88↓) Lymphocytes(%) 03.80 ± 0.58 03.60 ± 0.25 (5.26↓) 03.60 ± 0.25 (5.26↓) Eosinophils(%) * 02.40 ± 0.25 03.40 ± 0.25 (41.67↑) 03.20 ± 0.20* (33.33↑) Monocytes(%) (Data: Mean ± SEM, group. ↑ :- Increase,

The values in parenthesis are the percentage change in comparison to Normal Control ↓ :- Decrease, * :- P< 0.05 by student‟s „t‟ test for unpaired data)

Table – 3 : Effect of test drug on RBC and Platelet related parameters RBC and Platelet Group related Parameter NC S. asoca S. thaipingensis 7.58 ± 0.19 7.57 ± 0.18 (0.05↓) 7.66 ± 0.26 (1.11↑) Total RBC count(10e6/µl) 14.24 ± 0.32 14.80 ± 0.57 (3.93↑) 13.90 ± 0.33 (2.39↓) Haemoglobin (g %) 44.34 ± 1.14 44.22 ± 0.77 (0.27↓) 44.18 ± 1.13 (0.36↓) P.C.V. (%) 58.50 ± 0.53 58.44 ± 0.91 (0.10↓) 57.76 ± 0.75 (1.26↓) MCV (fl) 18.82 ± 0.20 18.78 ± 0.25 (0.21↓) 18.18 ± 0.28 (3.40↓) MCH (pg) 32.14 ± 0.45 32.14 ± 0.22 (0.00) 31.48 ± 0.24 (2.05↓) MCHC (g/dl) 3 1049.60 ± 67.01 1056.20 ± 62.54 663.00 ± 102.64*† (36.83↓) Platelet count (10e /µl) (0.63↑) (Data: Mean ± SEM, The values in parenthesis are the percentage change in comparison to Normal Control group. ↑ :- Increase, ↓ :- Decrease, * :- P< 0.05 by student‟s „t‟ test for unpaired data, † :- P< 0.05 by ONE WAY ANOVA & Dunnet‟s Multiple „t‟ test as post-hoc test.)

Table – 4 : Effect of test drug on BSL and lipid profile Parameter Group NC S. asoca S. thaipingensis *† 80.20 ± 4.76 100.00 ± 5.02 (24.69↑) 94.80 ± 5.72 (18.20↑) BSL (mg/dl) 65.20 ± 5.32 63.40 ± 2.16 (2.76↓) 67.40 ± 7.33 (3.37↑) S.Cholesterol (mg/dl) 89.25 ± 6.47 92.20 ± 3.73 (3.31↑) 106.25 ± 7.16 (19.05↑) S. Triglyceride (mg/dl) 38.80 ± 2.31 36.80 ± 2.75 (5.15↓) 33.00 ± 4.01 (14.95↓) HDL Cholesterol (mg/dl) (Data: Mean ± SEM, The values in parenthesis are the percentage change in comparison to Normal Control group. ↑ :- Increase, ↓ :- Decrease, * :- P< 0.05 by student‟s „t‟ test for unpaired data, † :- P< 0.05 by ONE WAY ANOVA & Dunnet‟s Multiple „t‟ test as post-hoc test.)

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Table – 5 : Effect of test drug on serum biochemical parameters serum biochemical Group Parameter NC S. asoca S. thaipingensis 54.60 ± 2.93 50.60 ± 1.50 (7.33↓) 65.60 ± 5.19 (20.15↑) Blood Urea(mg/dl) 0.62 ± 0.05 0.56 ± 0.07 (9.68↓) 0.58 ± 0.06 (6.45↓) S.Creatinine(mg/dl) 42.60 ± 3.86 47.60 ± 4.62 (11.74↑) 36.40 ± 3.96 (14.55↓) S.G.P.T.(IU/L) 126.20 ± 8.00 147.80 ± 11.16 (17.12↑) 121.60 ± 10.52 (3.65↓) S.G.O.T.(IU/L) 7.90 ± 0.19 7.80 ± 0.15 (1.27↓) 7.66 ± 0.39 (3.04↓) Total Protein(g/dl) 4.38 ± 0.07 4.38 ± 0.16 (0.00) 4.12 ± 0.26 (5.94↓) Albumin(g/dl) 3.52 ± 0.12 3.42 ± 0.10 (2.84↓) 3.54 ± 0.31 (0.57↑) Globulin(g/dl) 1.26 ± 0.03 1.26 ± 0.07 (0.00) 1.20 ± 0.13 (4.76↓) A/G ratio Alkaline Phosphatase(IU/L) 134.60 ± 32.19 192.40 ± 19.51 (42.94↑) 169.60 ± 38.65 (26.00↑) 0.38 ± 0.05 0.34 ± 0.04 (10.53↓) 0.34 ± 0.12 (10.53↓) Bilirubin(T)(mg/dl) 0.88 ± 0.09 1.26 ± 0.17 (43.18↑) 1.08 ± 0.20 (22.73↑) Uric Acid(mg/dl) (Data: Mean ± SEM, The values in parenthesis are the percentage change in comparison to Normal Control group. ↑ :- Increase, ↓ :- Decrease)

The effect of test drugs on RBC related parameters is shown in Table no. 3. Both the test drug treated groups did not show any significant changes in RBC related parameters. Test drug B decreases the Platelet count significantly in comparison to NC group, while the test drug S. asoca did not show any significant changes. Effect of test drug on BSL, lipid profile is presented in Table no. 4. Animals from S. asoca group exhibit significant elevation in the BSL while non-significant up & downs were observed in lipid profile in comparison to NC group. Test drug S. thaipingensis did not produce any significant changes in these parameters. Both the test drugs did not produce any significant change in Blood Urea, S. Creatinine, S.G.P.T., S.G.O.T. activity, Total Protein, Albumin, Globulin, A/G ratio, Alkaline Phosphatase activity & Bilirubin. (Table no. – 5) DISCUSSION Ashoka, which is an economically important plant, has become quite scarce in several localities and is reported to be threatened in North Eastern Region of India (Sharma PC et al., 2005; Alok Sharma, 2008).

Hence, it has become the target for adulteration of its bark. A drug is taken for the beneficial effect so it must not produce any toxicological changes in the recipient‟s body. But till date no study has been reported on the safety aspect of Ashoka Ksheerapaka of S. asoca and S. thaipingensis. 15 days after drug administration, no mortality was observed hence both the drugs are not lethal even at the limited test dose. Normal weight gain shows there are no serious effects on the body weight which was common side effect in synthetic estrogenic compound which were mostly used in menstrual disorders (Olinda rola, 2012). Both the drugs elevate the WBC count nonsignificantly and no significant changes were observed in Neutrophils percentage in comparison to NC group values i.e. both the drugs restricted the increase in WBC count and Lymphocyte percentage which was observed significantly in earlier studies on oral contraceptive pills. Further, Monocyte percentage in S. asoca group (41.64%) & S. thaipingensis group (33.33%) was significantly increased which is also in accordance with property of estrogens, this finding suggests that test drugs modulate the monocyte numbers and its effect may be mediated through estrogen receptor in monocytes (Sajida SH et al., 2006).

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Both the test groups did not produce any significant change in RBC related parameters i.e. RBC count, Hb, MCV, MCH, MCHC. Platelet count in S. thaipingensis group was decreased significantly (36.83%), while in S. asoca group no significant changes were observed in comparison to NC group values. It can be suggested that observed no significant effect on Hb in both the test drugs was in accordance with the earlier studies on oral contraceptive pills (Sajida SH et al., 2006). In S. asoca group significant increase in BSL (24.69%) was observed in comparison to NC group value while on lipid profile no significant changes were observed. S. thaipingensis did not produce any significant changes on glycemic control (BSL) value and on lipid profile. S. asoca lowers S. cholesterol (2.76%) and HDL cholesterol (5.15%) nonsignificantly in comparison to NC group values. Both the test formulations did not produce any significant changes in Blood urea, S. creatinine, SGPT, SGOT, Total protein, Albumin, Globulin, A/G ratio, Alkaline phosphatase, Bilirubin levels. This indicates that they do not have any toxicological

implication for acute administration even at very high dose levels. CONCLUSION Both the species of Saraca i.e. S. asoca & S. thaipingensis are safe at limited test dose when administered orally in the form of Ksheerapaka. However further toxicological evaluation like chronic toxicity studies etc. are required to provide complete safety profile. Both the drugs modulate the Monocytes percentage which may produce its estrogenic effect by affecting the estrogenic receptor on Monocytes which may be evaluated only after detailed efficacy related study on these plants. ACKNOWLEDGEMENTS The authors are thankful to the authorities of IPGT and RA, Gujarat Ayurved University for providing facilities to carry out the research work. One of the authors extends his deep gratitude to Director General, CCRAS, New Delhi & Dr. Sudesh Gaidhani, Dy. Director (Pharmacology) CCRAS, New Delhi, for providing fellowship.

REFERENCES Alok Sharma, Shanker C, Lalit Kumar Tyagi, Mahendra Singh, Ch.V.Rao (2008). Herbal Medicine for Market Potential in India: An Overview. Academic J. Plant Sci; 1 (2): 26–36. Anonymous (1986). The Ayurvedic Pharmacopoeia of India, part I, Vol. I. New Delhi: Ministry of Health and Family Welfare, Department of AYUSH, Govt. of India; pp. 14 Burtis CA, Ashwood ER, (eds) (1999). Tietz Textbook of Clinical Chemisry, 3rd edition, Philadelphia PA: Moss DW, Henderson AR; pp. 652.

Doumas BT, Arends RL, Pinto PC (1972). In: Standard methods of clinical chemistry. Chicago: Academic Press; Vol 7. pp. 175–89. Fossati

P, Prencipe L (1982). Serum triglycerides determined colorimetrically with an enzyme that produces hydrogen peroxide. Clin Chem; 28:2077–80.

Govind Das (1970). Bhaishajya Ratnavali, Streerogadhikara, 5/17–23, 114–116. In: Pandit Shree Lalchandraji Vaidya (ed.). Delhi: D.P.B. publications; pp. 698, 699, 704.

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 270–277

Kabasakalian P, Kalliney S, Wescott A (1973). Determination of uric acid in serum, with use of uricase and tribromophenolaminoantipyrine chromogen. Clin Chem; 19:522. Kirtikar KR, Basu BD (2001). Indian Medicinal Plants. In: Blatter E, Caius J, Mhaskar KS (eds), 2nd ed. Dehra Dun: Bishen Singh Mahindra Pal Singh Publishers; vol 9. pp. 3068–3069. Nalin (2005). Coment on PlantFiles: Red Saraca, Ashoka Tree, SorrowlessTree Saraca declinata cited, retrieved on 23.01.2013 from: http://davesgarden.com/guides/pf/go/67 517/ Olinda rola (2012). Estrogen Side Effects – Do You Know Them? retrieved on 17.08.2012 from: http://www.healthguidance.org/entry/28 52/1/Estrogen-Side-Effects---Do-YouKnow-Them.html PC Sharma, MB Yelne, TJ Dennis (eds) (2005). Database on medicinal plants used in Ayurveda, Vol. 3, New Delhi: Documentation and Publication Division, Central Council for Research in Ayurveda & Siddha; p76–87. Pearlman PC, Lee RT (1974). Detection and measurement of total bilirubin in serum with use of surfactants as solubilising agents. Clin Chemi; 20:447. Pennock CA, Murphy D, Sellers J, Longdon KJ (1973). A comparison auto analyzer method for the estimation of glucose in blood. Clin Chim Acta; 48:193–201.

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Pradhan P, Joseph L, Gupta V, Chulet R, Arya H, Verma R, Bajpai A. (2009). Saraca asoca (Ashoka): A Review. J. Chem. Pharm. Res.; 1(1):62–71. Roeschlau P, Bernt E, Gruber WA (1974). Enzymatic determination of total cholesterol in serum. Jour clinical Chem clinical Biochem; 12:226. Sajida SH, Sanaa MT, Amjad FA (2006). The effect of oral contraceptive pills on haematological indices, Tikrit Medical Journal; 12(1): 65–69. Tripathi Brahmanand (2004). Sharangdhar samhita,Sharangadhara, Madhyama khanda, Kwathadikalpana, 2/161. In: Dr. Brahmanand Tripathi (ed.). Varanasi: Chaukhamba Surabharati Prakashan; pp. 159. Shashikant Patwardhan (2013). Medicinal Herb: Ashoka [Saraca Indica], retrived on 23.01.2013 from: http://ayurvedaforyou.com/ayurveda_herb/ashok.html Slot C (1965). Plasma creatinine determination. A new and specific Jaffe reaction method. Scand J Clin Lab Invest; 17:381–87. Tietz NW (1986). Text book of Clinical Chemistry. W.B. Saunders; 579. Tietz NW (1995). Clinical guide to laboratory tests, 3rd ed. Philadelphia, PA: WB Saunders; pp. 76. Wilkinson JH, Boutwell JH, Winsten S (1969). Evaluation of a new system for kinetic measurement of serum alkaline phosphatase. Clin Chem; 15:487–95.

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Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

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