Indexing links of GJRMI GJRMI has been indexed in the Following International Databases Google Scholar, ProQuest, AYUSH RESEARCH PORTAL - Department of AYUSH, Ministry of Health & Family welfare, Govt. of India - http://www.ayushportal.ap.nic.in/
Index Copernicus - http://journals.indexcopernicus.com/karta.php?action=masterlist&id=7552 NewJour - http://old.library.georgetown.edu/newjour/nj2/msg29333.html Ulrich’s Periodicals Directory - http://ulrichsweb.serialssolutions.com/login This could be accessed only with a Login ID or through a Major academic Library at your place as it is a subscription datablase ScienceCentral - http://www.sciencecentral.com/site/4546968 getCITED - http://www.getcited.org/pub/103501008 Geneva Foundation for Medical Education & Research http://www.gfmer.ch/Medical_journals/Traditional_medicine_and_complementary_alternative_medicine.htm
Catalog ebiblioteca - http://cuhvl.c17.es/index.php/opac/action/default/?query%5Brevista%5D=^G*&query%5Banio%5D=&submitted=B%C3%BAsqueda&pageID=4
Ayurbhishak - http://ayurbhishak.wordpress.com/ayurvedank/ Medicinal plants (Dravya Guna) - http://indianmedicine.tripod.com/id30.html RoMEO - http://www.sherpa.ac.uk/romeo/pub/1312/ The following are a list of internationally reputed databases for which we have applied & it is under process…Links will be available after it gets done…
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ISSN 2277 – 4289 www.gjrmi.com Editor-in-chief Dr Hari Venkatesh K Rajaraman
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INDEX – GJRMI, Vol.1, Iss. 11, November 2012 Medicinal plants Research Nutrition & Food Sciences NORTH AMERICAN BIOACTIVE PLANTS FOR HUMAN HEALTH AND PERFORMANCE Ferreira Maria Pontes, Gendron Fidji, McClure Katrina C, Kindscher Kelly………………...568–582 Bio-technology PREDICTION OF MOLECULAR TARGET OF SISSOTRIN USING DOCKING TECHNIQUES Zaman Aubhishek, Shafrin Farhana…………………………………………………………...583–598 Ethno-Botany PHYTOCHEMICAL ANALYSIS AND ANTIOXIDANT ACTIVITY OF THREE ETHNO MEDICINAL PLANTS OF PACHAIMALAIS, TIRUCHIRAPPALLI DISTRICT, TAMIL NADU. Rekha S, Parvathi A….………………………………………………………………………..599–611 Public Health NEED AND RELEVANCE OF FORMATION OF INDIAN SYSTEMS OF MEDICINE AND HOMOEOPATHY (ISM & H) POLICY 2002 IN INDIA Singh Balpreet, Kaur Rajvir, Kumar Manoj, Singh Amarjeet………………………………...612–619
Indigenous medicine Ayurveda PHARMACOLOGICAL STUDY TO ASSESS THE VIPAKA OF CERTAIN SAMANA & VICITRA PRATYAYARABDHA DRUGS IN ALBINO RATS Jadoun Anuruchi, Solanki S K, Ashok B K, Dwivedi R R…………………………………...620–628 PARKINSONISM IN AYURVEDIC PERSPECTIVE, A BIRD’S EYE VIEW Nayak Annada Prasad…………………………………………………………………………629–638 SCOPE OF THIN LAYER CHROMATOGRAPHY IN QUALITATIVE EVALUATION OF AYURVEDA DRUGS Rajendra H M, Lad Meenal D…………………………………………………………………639–643 A REVIEW ON PHARMACODYNAMICS OF VAMANA KARMA AND VAMANOPAGA DAŚEMĀNI Ujjaliya Nitin, Rema devi R…………………………………………………………….……..644–649
COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – LEAF OF ALBIZIA LEBBECK (L.) BENTH, LEGUMINOSAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT, KARNATAKA, INDIA
Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 568–582 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Review article NORTH AMERICAN BIOACTIVE PLANTS FOR HUMAN HEALTH AND PERFORMANCE Ferreira Maria Pontes1*, Gendron Fidji2, McClure Katrina C3, Kindscher Kelly4 1
Assistant Professor, Department of Nutrition and Food Science, Wayne State University, 5045 Cass Avenue, Science Hall 3009, Detroit, MI, USA, 48202 2 Assistant Professor, Department of Science, First Nations University of Canada, 1 First Nations Way, Regina, Saskatchewan S4S 7K2 Canada 3 Graduate Research Assistant, University of Kansas, Department of Geography, Lawrence, KS 66047 4 Senior Scientist, Kansas Biological Survey, University of Kansas, 2101 Constant Avenue, Lawrence, KS 66047 USA *Corresponding Author: Email: mpferreira@wayne.edu; Phone: (313) 577-5888; Fax: (313) 577-8616
Received: 20/09/2012; Revised: 25/10/2012; Accepted: 31/10/2012
ABSTRACT Native and naturalized bioactive plants of the Canadian and American temperate biome are examined for their health and performance enhancement properties. Some of these plants are now being used as natural health products, and many have a long history as traditional foods and/or medicines with indigenous groups. This paper reviews the medicinal/cultural uses and bioactive properties of selected plant families: the Holly family (Aquifoliaceae) as stimulants, the Celery family (Apiaceae) as normoglycemic aids and analgesics, the Ginseng family (Araliaceae) as energyboosting aids, the Sunflower family (Compositae) as anti-inflammatory aids, and the Legume family (Fabaceae) and Nightshade family (Solanaceae) as functional foods. These North American plants show promising avenues for innovative health and performance enhancement aids and it is concluded that they should be investigated further for their bioactive properties. KEY WORDS: Athlete; botanicals; complementary & alternative medicine; dietary supplement; ergogenic aid; grasslands; functional food; indigenous; natural health product; prairie; phytotherapy
To Cite this article: Ferreira M P, Gendron F, McClure K C, Kindscher K (2012), NORTH AMERICAN BIOACTIVE PLANTS FOR HUMAN HEALTH & PERFORMANCE, Global J Res. Med. Plants & Indigen. Med., Volume 1(11), 568–582
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 568–582
INTRODUCTION North Americans are avid users of complementary and alternative medicine (CAM), such as dietary supplements and natural health products (NHP), for health risk reduction and performance enhancement. Regarding performance enhancement, the American Dietetic Association, Dietitians of Canada, and the American College of Sports Medicine have a joint position statement on the use of supplements and ergogenic aids, due to the popularity of their use amongst athletes and fitness enthusiasts (American Dietetic Association et al., 2009). Regarding other consumer group use of botanicals, in 2007, approximately 40% of surveyed USA adults used CAM, and 18% of these therapies are dietary supplements (Barnes et al., 2008). In Canada, 71% of adults surveyed in 2005 have used NHP (Ipsos, 2005). CAM is a broad group of diverse medical and health care systems (e.g., traditional / indigenous medicine), various practices (e.g., sweat lodge, meditation), and products (e.g., functional foods, supplements, and NHP). Natural health products are naturally occurring substances derived from organisms from the 5 life kingdoms: plants, animals, microbes, fungi, and protists. These products come in a variety of forms such as powders, extracts, ointments, capsules, and tablets. They include vitamins and minerals, botanical remedies, zootherapies, ergogenic aids, probiotics, homeopathic, and traditional medicines. Many traditional/indigenous food and medicines inform the research and development of relevant bioactive components into commercial NHP. Over 80% of Canadians believe that it is important to respect the role that NHP play in some cultures (Ipsos, 2005). Thus, traditional and indigenous knowledge systems continue to have a broad impact on the use of CAM by mainstream consumers. Across Canada and the USA, a large temperate grasslands biome (3 million km2) is host to a rich diversity of landscapes yielding a variety of native and naturalized plants. As
well, this biome boasts a rich cultural diversity, including dozens of indigenous groups with a long history of use of botanicals for food and medicine. Despite being one of the most human altered landscapes on the planet, due to colonialism and agriculture, the grasslands biome continues to be a source of functional foods and medicines for indigenous and nonindigenous people today (Kindscher, 1987, 1992). While CAM is not evidence-based, and thus not part of allopathic medicine, this is changing. Increasingly, there is growing scientific evidence documenting the potential health and performance value of North American traditional foods and medicine products. For example, Echinacea species (family Compositae) are wild perennials used widely in North America by indigenous people for a variety of health and performance purposes. Today, Echinacea species are one of the most commonly utilized NHP by mainstream North Americans; as well, it is one of the most researched botanicals from the Americas (Price and Kindscher, 2007). The National Institutes of Health (NIH) invested nearly $90 million in mechanistic, preclinical and clinical research studies of botanicals through two rounds of the Botanical Research Centers Program between 2000– 2010. The center is currently in its third fiveyear funding cycle. The Office of Dietary Supplements and National Center for Complementary and Alternative Medicine (both NIH entities) jointly orchestrate a network of interdisciplinary centers devoted to the study of botanicals, with the Botanical Research Centers Program being the largest (Coates and Meyers, 2011). In Canada, the functional foods and NHP sector continues to grow, with total annual revenues of $3.7 billion CND, as estimated from the Functional Foods and Natural Health Products Survey 2007 (Statistics Canada, 2007). Growth of this Canadian sector is enhanced through the spending of $148 million CND (in 2007) for research and development by functional food firms and those producing and/or marketing NHP. New developments of this wide range of product lines (over 22,000) include extraction
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and processing of bioactive compounds to produce products with increased health and well-being benefits. In this paper we give a brief introduction to the basic secondary metabolites in botanicals, and how they may be used for health and performance enhancement. We will then introduce selected native plants, organized by family, that come from a variety of habitats across the grasslands biome and have a history of utilization by indigenous people (past and/or present). Ethnobotanical and evidence-based information regarding the health and performance properties of the plant will be discussed with implications for future research. It is our hope that this multidisciplinary conversation regarding these plants will inspire indigenous interest in higher education; protection and appreciation of the varied habitats that occur in the North American grasslands biome; appreciation of the influence of traditional/indigenous knowledge on NHP use for health and performance; and scientific study of these bioactive plants. SECONDARY METABOLITES Plants produce many metabolites from secondary metabolism that may have clear roles in plant physiology (Wink, 2010). These secondary metabolites are organic molecules produced in response to influences such as developmental stage and environmental assaults (e.g., maturation and insects). When applied externally or internally, some of these plant compounds have purported beneficial effects in mammalian physiology. For example, as medicinal agents in folk remedies, veterinary medicine, and pharmacognosy, natural products continue to have significant medicinal, economic, and ecologic functions (Hopkins and Huner, 2009). There are 4 major classes of bioactive secondary metabolites: terpenes, phenolics, glycosides, and alkaloids. Terpenes are a diverse group of isoprenoid compounds with over 15,000 structures, thus forming one of the largest phyto-chemical groups. While many terpenes are primary
metabolites with significant plant growth and development roles (e.g., colour pigments, hormones, and sterols), the vast majority are secondary metabolites with roles in plant defense (e.g., essential oils and latex). Conifers and many flowering plants produce essential oils rich in terpenes and terpenoids, which may have medicinal properties. The native North American hops (Humulus lupulus L., Cannabaceae) are herbaceous perennial vines that grow in moist fertile soils in the prairie bioregion. The female flower clusters are valued by indigenous people for their sedative qualities and are used as an herbal analgesic and for insomnia. In the brewing industry, hops contribute anti-microbial, flavour, and aroma properties attributed in part to the terpene compounds. Beer has been used by athletes for its therapeutic qualities since recorded history; yet in immoderate doses alcohol continues to be abused amongst athletes (Ferreira and Willoughby, 2008). Polyphenols are a diverse, large group of simple or complex phenolic compounds derived from secondary metabolism of the aromatic amino acids. Downstream product groups include coumarin, lignin, flavonoids, tannins, and alkaloids. Most of these compounds have a role in chemical defense, some are metabolic end products with unknown function, and lignin is a major structural component of vascular plant cells. Lignin has emerged as an important component of dietary fibre (undigested organic polymer) with antioxidant potential and suppresses carcinogenesis in vitro (Fardet, 2010) likely by acting as an absorbent for potentially carcinogenic molecules in the digestive tract in vivo. The Academy of Nutrition and Dietetics endorses consumption of a high fibre diet (e.g., fruits, vegetables, and whole grains) for good health (American Dietetic Association, 2008). Glycosides are a class of compounds characterized by glycosidic bonds between a sugar molecule and another non-carbohydrate molecule with a hydroxyl group. This feature gives the compound the property to act as a detergent with a role in plant defense (e.g., anti-
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fungal). The four main groups of glycosides are saponins (have foaming properties), glucosinolates (anti-oxidants in cruciferous vegetables), cyanogenic glycosides (contains cyanide group), and cardiac glycosides (used to treat heart conditions). Although not native, the common dandelion, (Taraxacum officinale Webb, Compositae) is used in Mexican and Native American traditional medicine for a variety of ailments and health benefits (Rodriguez-Fragoso et al., 2008). Glucosides (glucose-derived glycosides) from this plant have been shown to exhibit in vitro antiinflammatory action as well as anti-oxidant action (Rodriguez-Fragoso et al., 2008). Alkaloids are a diverse group of chemically unrelated nitrogenous organic compounds with high biological activity. Herbaceous dicots (flowering plants) are rich in alkaloids, and many are repellant to vertebrates and invertebrates, likely due to their bitter taste. Many commercially important drugs are alkaloids, and most interfere with neurotransmitters (Hopkins and Huner, 2009). Thus, they often function as painkillers, muscle relaxers, and also as mood-altering substances. Caffeine, an alkaloid xanthine derivative, is one of the most widely used nutraceuticals in the world; and its use amongst athletes is similarly widespread and well-studied (Tarnopolsky, 2010). Common dietary sources of caffeine include coffee (typically Coffea arabica L. and C. canephora Pierre ex A. Froehner, Rubiaceae), mate (Ilex paraguariensis A. St.– Hil., Aquifoliaceae), tea (Camellia sinensis (L.) Kuntze, Theaceae), cocoa (Theobroma cacao L., Malvaceae), and energy drinks. Currently not a banned substance by the International Olympic Committee, caffeine is wellestablished as a potential aid to physical and mental performance. However, not all people are caffeine responders, and the effects of dietary caffeine are contingent upon whether a person is a habitual or a ‘naïve’ user. Studies that appropriately control for confounding factors indicate that psychomotor and moodenhancement benefits of caffeine use in
habitual users are likely due to a reversal of withdrawal symptoms (James and Rogers, 2005). According to a systematic review of the literature by Tarnopolsky (2010), caffeine has demonstrated adrenaline/noradrenaline receptor antagonistic effects upon skeletal muscle, adipose, and central nervous system tissues. Based upon his expert assessment of the currently available evidence, dietary doses of caffeine consumed prior to (2–6 mg/kg) or during (0.75–2 mg/kg) endurance exercise result in improved performance. While a reduction in central fatigue is a well-established benefit upon caffeine administration during endurance activities, caffeine does not significantly affect performance in highintensity or strength activities. HOLLY FAMILY (AQUIFOLIACEAE) The presence of xanthines (caffeine, theobromine, and methylxanthine) is restricted to a few species of the Ilex genus in the Americas. Indigenous groups consume mate (I. paraguariensis A. St.-Hil.), guayusa (I. guayusa Loes.), and yaupon (I. vomitoria Aiton). The first two are native to South America, and the latter to North America. Mate is used by the South American Guarani for socializing, mental and physical stimulation, and in herbal remedies. In southern South American culture, the consumption of mate rivals that of coffee as a mainstream stimulant. Of the 3 hollies, mate has been subject to the most scientific scrutiny and found to have compelling chemical profiles with important health implications (Heck and de Mejia, 2007). Guayusa is anecdotally used by the Amazonian Runa people as a balanced stimulant that energizes the body and mind while promoting restful sleep with lucid dreams. Consumed in social ceremony, the Runa appreciate mate’s capacity to decrease bodily pain and to increase mental clarity. Yaupon, or the yaupon holly (also cassina) was used by Southeastern USA bands where it grew, and by Texas plains bands who traded for it (Browne, 1935; Havard, 1896). Records indicate that it was used as a
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mildly stimulating infusion for frequent use, or more famously, as part of the emetic ‘black drink’ associated with ritual purging (Havard, 1896). Yaupon (Spanish: Acebo de Yupon) Yaupon, or cassina, is a native North American member of the Holly family. This evergreen perennial is native to the southeastern coastal plains of North America and the piney woods and post-oak savannah of Texas (Smith and Rechenthin, 1964) and naturalized west to central Texas. It may be native or naturalized to some parts of Mexico. It is a dioecious shrub or small tree with a height of 3–6 m, and distributed across a variety of soil types and light intensities. However, it prefers moist acidic soil and has intermediate shade tolerance. Aerial plant parts (e.g., roasted leaves) are caffeine-rich and were used by indigenous groups in ceremony (e.g., as the ‘black drink’ decoction) and as a phytomedicine (e.g., as an infusion). Natives called I. vomitoria Aiton, “cassina” (derived from the Timucua language) and I. cassine L. (commonly known today as dahoon holly) as “yaupon” and the black drink may have involved a mix of several species of Ilex (Power and Chestnut, 1919; Edwards and Bennett, 2005). The tangled nomenclature has been resolved, although confusion persists in the older literature (Edwin, 1963). In comparing the xanthine profiles of the two species, it has been demonstrated that caffeine and methylxanthine concentrations are higher in the yaupon than in the dahoon holly (Edwards and Bennett, 2005). It is also established that xanthine alkaloids (caffeine and theobromine) are present in the leaves of yaupon tree dependent upon soil nitrogen content (Palumbo et al., 2007). Roasting the leaves facilitates solubilization of caffeine, as does the steeping in hot water. Yet, published quantification of the caffeine content of ‘a cup of yaupon’ is lacking, and cannot currently be compared to a cup of coffee, mate, or tea. Aqueous infusions of the plant are rich in phenolic acids (dicaffeoylquinic acids) and
flavanols (quercetin and kaempferol glycosides) (Mertens-Talcott et al., 2011). Regarding the potential scientific merits of this plant for use as an ergogenic aid or nutraceutical, further work is encouraged. Yaupon holly has been analyzed by highpressure liquid chromatography (Edwards and Bennett, 2005; Mertens-Talcott et al., 2011; Palumbo et al., 2007), and discerned to have a number of phenolic compounds, as well as alkaloid xanthenes (Edwards and Bennett, 2005; Palumbo et al., 2009). Principal compounds identified by mass spectrometry are alkaloids and some phenols (Mertens-Talcott et al., 2011). Many of these secondary metabolites likely protect the plant against bacterial and fungal microbial overgrowth. There are no scientific reports on the health properties of yaupon for humans; but Noratto and colleagues (2011) indicate antiinflammatory and anti-oxidant activity for the polyphenol (quercetin and kampferol glycosides) extractions of the plant in vitro. They demonstrated that yaupon-derived flavanols inhibit human colon cancer cell viability in vitro through reduced inflammation by induction of endogenous anti-oxidant systems and through reduced activation of proinflammatory genes. While yaupon holly has an intriguing profile of bioactive compounds, when used as a NHP, the anti-tumour, antiinflammatory, and anti-oxidant properties have yet to be demonstrated in humans. CELERY FAMILY (APIACEAE) There are nearly 3,000 members in the Celery family (Apiaceae) worldwide (Bidlack et al., 2010). These are mostly temperate plants with soft-hollow stems and the leaves are usually alternate and pinnately or palmately compound with white umbellate inflorescences. The seeds and fruit form below the point of origin of the petals and stamen; they vary in size and shape and many have culinary and/or medicinal uses. Commonly used seeds include: dill (Anethum graveolens L.), coriander (Coriandrum sativum L.), celery (Apium
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graveolens L.), cumin (Cuminum cyminum L.), anise (Pimpinella anisum L.), caraway (Carum carvi L.), parsley (Petroselinum crispum (Mill.) Mansf.), and fennel (Foeniculum vulgare Mill). The root in many species is edible (e.g., carrot (Daucus carota L.), parsnip (Pastinaca sativa L.), as well as the stems and leaves of celery, parsley, cilantro, and lovage (Levisticum officinale L.). These plants are aromatic due to essential oils rich in bioactive compounds that contribute to their use by people for food and medicine. Caution should be utilized when wild-harvesting members of this family because some are deadly poisonous (e.g., water-hemlock (Cicuta maculata L.). Oshá (Spanish: chuchupate) Oshá (Ligusticum porteri, Apiaceae) and other Ligusticum species are herbaceous perennials found from British-Columbia and further south into northern Mexico, at elevations of 500–3,500m. It grows in moist soils on the edge of wooded habitats, as well in drier, rocky soils. The roots are fibrous tubers with ‘hairy’ root crowns growing from a central rootstock. Below the dark brown outer ‘skin’, the off-white inner root has a distinctive and aromatic ‘celery’ aroma. Animals and people consume the leaves, seeds, and roots as food and medicine. Many indigenous and traditional people anecdotally refer to oshá as ‘Bear Medicine’, because bears utilize the root for food and medicine (Andrews, 1992; Lipske, 1993). Currently, oshá is widely used by Latin and Native Americans, as well as by mainstream NHP consumers. Cultivation of oshá has been unsuccessful and is currently wild-harvested. Over-harvesting may be a concern, and is being studied by researchers at the Kansas Biological Survey, University of Kansas, USA. Currently, it is sold in Canada, the USA, and Mexico in the form of tinctures, capsules, whole or ground roots, and seeds. The roots and seeds have been infused as traditional and folk medicine, for a variety of ailments; indigenous runners anecdotally chewed the root for stamina and power. Other species of the Ligusticum genus,
native in Asia, are widely used in Chinese Traditional Medicine. As a root infusion, oshá is used to treat diabetes (Andrade-Cetto and Heinrich, 2005). Type 2 diabetes is the world’s most common endocrine disorder, with an escalating incidence especially amongst the Native and Latin American populations. A recent study reports the hypoglycemic effect of oshá extract in streptozotocin diabetic mice (Brindis et al., 2011). Several compounds isolated from the extract utilized in that study proved to have anti-hyperglycemic activity in vivo, with the mechanism of action being undetermined or due to inhibition of intestinal alphaglucosidase. One compound isolated from the extract has previously been shown to stimulate insulin secretion. Thus, the methanol root extract of oshá represents a normoglycemic agent with bioactive compounds with differing mechanisms of action, worthy of further scientific investigation. Tincture made from soaking the oshá root in ethanol for months is commonly used as an externally applied liniment for sore muscles. The search for novel analgesics continues, because many common analgesics have adverse side-effects. Botanical anti-nociceptive substances recently discovered include alkaloids, terpenoids, and flavonoids. While chemical analysis of other Ligusticum species (e.g., Asian species) has indicated the presence of furano-coumarins, pyrano-coumarins, and phthalides, a composite chemical profile on oshá, reported by Brindis and colleagues (2011), indicates the presence of phenylpropanoids, terpenoids, and phthalides. These phyto-chemicals have yet to be specifically analyzed for anti-nociceptive properties. However, orally administered methanol-choloroform extracts of oshá in mice subjected to the writhing test study resulted in significant decrement of pain-related behavior (Deciga-Campos et al., 2005). These preliminary findings suggest a place for this plant in analgesic discovery research.
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GINSENG FAMILY (ARALIACEAE) Panax species of the Ginseng family Araliaceae are found native in Asia and North America. These perennials have woody or herbaceous species with umbel flowers. The genus name, Panax is derived from the Greek word panacea. Panax is a part of the subfamily Aralioideae and the 3 species of the genus recognized as medicines are: Asian ginseng (P. ginseng C.A.Mey.), Japanese ginseng (P. japonicas (T.Nees) C.A.Mey.), and American ginseng (P. quinquefolius L.). The most commonly known family member is the Asian ginseng, which is used worldwide as a NHP. The indigenous people in New France used the American root to stimulate appetite and to treat rheumatism and dysentery. Colonists shipped this root to Asia, which proved to be profitable, as the Chinese considered the plant valuable (Messier, 1989). While harvest of wild American ginseng is legal during statespecified periods in the USA, it is illegal in Canada (COSEWIC, 2000). The English name ginseng derives from its Chinese name, rénshēn, meaning "man root" because the roots are often shaped like human legs. Panax spp. contain over 150 ginsenosides, the bioactive compounds that are characteristic of true ginseng. Ginsenosides are a subclass of the triterpenoid saponin glycosides (Senchina et al., 2009). Ginseng is commercially available as a NHP in dosage forms suited for oral administration. In a review by Senchina and colleagues (2009), it was found that, along with echinacea, (Echinacea angustifolia DC., Compositae), Asian ginseng is the most popular NHP used by athletes, with 3.2–15% of athletes using it. Athletes use NHP for health and performance more than other consumer groups. Athletes consume members of the genus Panax for their alleged increase in energy and physical stamina. Although members of the genus Panax have been reported to increase pulmonary function and exercise capacity and to reduce chronic fatigue in patients and elderly people, there is no evidence of these effects in healthy, young athletes (Bahrke et al., 2009). In
a review by Bahrke and colleagues (2009), it is also reported that members of the genus Panax do not improve performance and recovery of individuals undergoing exhaustive exercise. Asian ginseng supplementation taken one hour prior to exercise tests on a treadmill did not affect the endurance running performance and other selected physiological parameters in recreational runners (Ping et al., 2011). Senchina and colleagues (2009) conclude that the benefits of taking Asian ginseng supplements on athlete immune system remain largely unproven. Small Spikenard (French: aralie à tige nue (Marie-Victorin, 1964); Anishnabe: jiisens ojiibikan gisēns, wenane) Small spikenard (Aralia nudicaulis L., Araliaceae), commonly referred to as wild sarsaparilla, is a characteristic feature of parkland groves and wooded ravines of the prairie grasslands except the extreme South and Southwest USA (Vance, 1999). This perennial member of the family Araliaceae, genus Aralia, has single leaf stalks, 15–30 cm in length before dividing into 3 parts that then divide again into leaflets. The greenish white flowers are borne on a flowering stalk, which is usually below the level of the leaves. The plant was used as a popular medicine in New France, but it was only during the English-controlled period that the plant was harvested on a large scale. For example, it is reported that several thousands of pounds were sent to London merchants in 1766 (Lessard, 1996). Indigenous Elders from Saskatchewan often refer to small spikenard as the energy plant, and also as rabbit root. Consistent with other Ginseng family members, the wild sarsparilla root is used as a tonic and stimulant, although not to the same extent as commercial Asian ginseng. The spongy root pith has a sweet balsam aroma and flavour and indigenous people use the root and berries as a stimulating tonic on long journeys (Keane and Howarth, 2009). In Saskatchewan, some pubs still brew sarsaparilla root beer, which is made from
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natural sarsaparilla flavour (indeterminately of wild sarsaparilla vs. non-native source). Brussel (2004) also mentions its stimulant effect. Its high carbohydrate level might explain its energetic quality. In a study of 17 species of plants growing in the southern edge of the boreal forest in Ontario, Canada, small spikenard fruits showed the greatest total energy value (73.4 Kcal/100 g) and soluble carbohydrate based upon glucose (16.38%) (Usui et al., 1994); the fat and protein composition of its fruits was 0.28% and 1.34%, respectively. While studies on the role of small spikenard as an energy source have been limited, its ethnobotanical history and nutritional value suggests the need for further examination for its use as a NHP. There have been several published and ongoing mechanistic and clinical studies of ginseng and NHP. They show promise to be consistent, safe and effective in a variety of health-related areas of research. Regarding wild sarsaparilla, a small spikenard rhizome extract was found to have little anti-mycobacterial activity on its own but appeared to have some synergistic activity with Bacillus Calmette– Guérin (tuberculosis vaccine) and Mycobacterium avium when combined with Symplocarpus foetidus (Webster et al., 2010). Rhizome extracts (hexane) were effective at eliminating 4 different human cancer cell lines with cellular viability less than 6.8% (Huang et al., 2006). In a companion study, Wang and colleagues (2006) reported that hexane extraction from the rhizome and the fruit of small spikenard were more effective than stem and leaf extraction against human colon cancer, leukemia, and cervix cancer in cell lines. SUNFLOWER FAMILY (Compositae) The family Compositae has approximately 20,000 species and is distributed worldwide (Bidlack et al., 2010). Well-known members of this family include chicory (Cichorium intybus L.), Jerusalem artichoke (Helianthus tuberosus L.), Texas tarragon (Tagetes lucida Cav.), sunflower (Helianthus annuus L.), and prairie
thistle (Cirsium canescens Nutt.). The edible tubers of Jerusalem artichokes contain a starch comprised of fructose polymers. Roasted dandelion and chicory roots have been used as a coffee substitute. Sunflower, native to America, was widely used by indigenous people for their seeds. Yarrow (French: achillée millefeuille; Anishnabe: aadjidamowana waabanowashk zhagish kaandawens aanashic) Yarrow (Achillea millefolium L., Compositae) is one of the most abundant white flowers growing upon the North American prairie. It has white flower heads densely packed in a round-topped terminal cluster. Its woolly leaves are divided into many segments and grow from a branched rhizome. One Elder in Saskatchewan calls this species porridge-ona-stick and states that an infusion “made using the entire top of the plant helps support the immune system and can be used for chest infection” (Gendron et al., 2009). Indigenous people in Saskatchewan use this plant for regulating body temperature, opening skin pores, stimulating perspiration, and treating colds and fevers. Yarrow is also used to regulate the menstrual cycle, heal tissues, and to reduce inflammation (Keane and Howarth, 2009). Recently, it was found that methanol extract of yarrow inhibited human neutrophil elastase and matrix metalloproteinases MMP-2 and -9 in vitro (Benedek et al., 2007). These enzymes are proteases associated with the inflammatory degradation of the connective tissue and the extracellular matrix proteins. Benedek and colleagues (2007) were unable to identify which flavonoids contributed the most to the neutrophil elastase inhibition. The capacity of yarrow to reduce inflammation may be an interesting avenue to explore in regards to mechanical stress associated with exercise. Yarrow extracts exhibit blood-pressure lowering (in vivo & in situ), vaso-dilatory (in vitro) and broncho-dilatory (in vitro) activities (Khan and Gilani, 2011).
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Yarrow’s high content of flavonoids makes it a potential candidate for chemoprevention research. Dried aerial parts of yarrow contain flavonoids, alkaloids, coumarins, saponins, sterols, tannins, and terpenes. Infusions made with yarrow are a source of the flavonoids rutin, chlorogenic acid, and quercetin (Dadáková et al., 2010). Yarrow has a high amount of apigenin, a flavonoid associated with cancer prevention (Patel et al., 2007). Activities identified for apigenin-mediated cancer prevention and therapy include: estrogenic/antiestrogenic; anti-proliferative; cell-cycle arrest and apoptosis; anti-oxidation; detoxification enzyme induction; immune-protection; and cell signal modulation (Patel et al., 2007). The Achillea genus also shows promising potential at reducing incidence of degenerative diseases, such as atherosclerosis, with its high antioxidant activity (Vitalini et al., 2006). LEGUME FAMILY (LEGUMINOSAE) The Legume family (also Fabaceae) is the third largest plant family, with over 19,000 species (Bidlack et al., 2010). The family name Fabaceae refers to the fruit, which is also called a legume or pod. This large and economically important family is characterized by having many species that form symbiotic relationships with bacteria to fix atmospheric nitrogen, resulting in plants with rich nitrogen content, and hence in protein. The many important plants in the Legume family world-wide include varieties of beans (Phaseolus vulgaris L.), soybeans (Glycine max (L.) Merr.), peas (Pisum sativum L.), cowpeas (Vigna unguiculata (L.) Walp.), peanuts (Arachis hypogaea L.), chickpeas (Cicer arietinum L.), alfalfa (Medicago sativa L.), and carob (Ceratonia siliqua L.). The largest genus is Astragalus with over 2,000 species; many of which are medicinal while many others are poisonous due to alkaloids, selenium accumulation, or both (Kindscher, 1987). Prairie Turnip (Lakota: tipsin) The prairie turnip (Pediomelum esculentum (Pursh) Rydb.), also called Indian bread root or
bread-root scrufpea, is a perennial prairie plant found throughout the grasslands from Alberta to Texas. The scientific name tells one it is the esculent or edible species of the genus. It has a chicken egg-sized swollen root that appears turnip-like, but tastes more like a potato with a hint of peanuts. This was the most important wild plant food collected by indigenous people across the Great Plains of North America (Kindscher, 1987; Nabhan and Kindscher, 2006). The roots are often braided by the tap roots and are still traded and stored by the Lakota and other indigenous bands. The longlived plants have annual growth rings and are found today only on native prairies. The plants occur in patches and historically, large prairie turnip patches were places where people went to dig them during the early summer before the tops dried up and blew away as tumbleweeds. The Omaha were known to determine the route of their summer buffalo hunts by where they could camp and harvest an abundance of prairie turnips (Fletcher and La Flesche, 1911). The harvest was an event for the women and children, with the kids seeking new plants as a game while the women used digging sticks to harvest them from the hard prairie soil (Gilmore, 1977). Their abundance was evident as observed in 1858, as the Cree in Saskatchewan were observed to harvest “many bushels” (Mandelbaum, 1940). Lewis and Clark recorded on May 8, 1805 in northern Montana that “this root forms a considerable article of food with the Indians of the Missouri...they are esteemed good at all seasons of the year...are sought and gathered by the provident part of the natives for their winter store, when collected are stripped of their rind and strung on small throngs or chords and exposed to the sun or placed in the smoke of their fires to dry; when well dried they will keep for several years” (Thwaites, 2001). The native harvesters of these plants came back to their prairie turnip patches repeatedly over the years, but not every year. Fortunately, when the plants are harvested, the seeds are ripe, and traditionally these were scattered across the disturbed soil as the tops of plants
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were returned to the hole from which they were harvested. Research has shown that disturbance surrounding prairie turnips increases the numbers that are present (Castle, 2006) and helps justify the sustainability of traditional harvest and planting practices. The nutritional profile of the prairie turnip indicates that in addition to starch, it has high levels of calcium, magnesium, and iron. Also the prairie turnip is a good source of the amino acid lysine and is considered an “exceptionally valuable dietary supplement” (Kaldy et al., 1980). It would seem that the prairie turnip could be suited as a functional food. It has been difficult to cultivate, however, and it is not readily available in the commercial trade. NIGHTSHADE OR POTATO FAMILY (SOLANACEAE) The Solanaceae are well known as an important group of plants that are used in food and medicine and contain alkaloids whose toxins range from very mild to very poisonous. Phyto-chemical research has been occurring for approximately 200 years. Members of this family are particularly interesting for their competitive characteristics, high productivity, and a notable presence of secondary metabolites (Eich, 2008). Steroidal alkaloids or alkamines are present such as tomatillidine from Solanum tomatillo (Remy) Philippi f. (Eich, 2008). Tropane alkaloids are found in the Physalis genera amongst others in the family. Steroidal and tropane alkaloids are nonoverlapping in taxa and are thus mutually exclusive (Wink, 2010). Wild Tomatillos (Zuni: k’ia’-po-ti-mo’-we) The wild tomatillo, or ground-cherry, (Physalis longifolia Nutt.) is a low-growing perennial herb of weedy habitats. The papery husk that encloses the fruit of the wild tomatillo is a distinctive characteristic of the genus Physalis, which includes cultivated species such as husk-tomato (P. philadelphica Lam.) and Chinese lantern (P. alkekengi L.). The name Physalis is Greek for “a bladder,” a
reference to the inflated calyx, which forms the husk. The genus Physalis is a member of the Nightshade family, Solanaceae, which includes tomatoes (S. lycopersicum Lam.), potatoes (S. tuberosum L.), and tobacco (N. tabacum L.). All nightshades are considered somewhat poisonous and may contain toxins in some parts of the plant, but many fruits in the family Solanaceae are edible, including wild tomatillos (Kindscher, 1987). The many-seeded berries of the genus range from greenish to yellow to tangerine and are sometimes flushed with purple or red (Whitson and Manos, 2005), thus explaining the use of the common names “ground tomato” and “husk tomato” with reference to Physalis (Castetter, 1935). Wild tomatillos occur throughout the continental U.S. and into southern Canada and northern Mexico. Its habitat includes old fields, open woods, and prairies, but it thrives in disturbed sites, including roadsides. Plants form colonies through the spread of underground rhizomes. The widely distributed wild tomatillo is cultivated by Zuni women, who boil the ripe red berries before grinding them with raw onions, chile, and coriander seeds in a mortar. The dish is regarded as a great delicacy (Kindscher et al., 2012; Castetter, 1935). Ethnologist Walter Hough (1898) stated that in the “old times”, the berries were eaten by the Hopi. Hough (1898) also reported that the Zuni dried and ground the berries to produce a meal for making bread. According to Matilda Cox Stevenson (1915) the berries of the ivy-leaf ground-cherry (P. fendleri A. Gray, now recognized as P. hederifolia A. Gray) had the same Zuni name (Ke’tsitokia) as this wild tomatillo (and is named for an insect that feeds upon the plant), indicating that these species may have been used interchangeably. She reported that this plant grows wild on lowlands and is also cultivated in the small gardens worked by women. In Frank Cushing’s (1920) Zuni Breadstuffs, he states that: “Among the sandy defiles of the upper plains, mesas, and mountains grow abundant low bushes bearing very juicy little yellow berries called k’ia’-po-
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ti-mo’-we, or the “juice-filled fruitage.” These berries were in high favour with the ancient Zuni as food. They were boiled or stewed to make a sweet but acrid sauce, which although not quite so acidic, otherwise resembled the cranberry. The berry is still used as food at the Zuni Reservation. In Rita Edaakie’s (1999) “Idonaphshe” -let’s eat- traditional Zuni foods” she gives a recipe for the use of K’e:ts’ido’kya K’yalk’osenne or tomatillo paste in which the store purchased husk-tomatoes or tomatillos (P. philadelphica Lam.), are used to make a tasty sauce which includes roasted chilis and green onions, as an adaptation of the previous use of native Physalis species. Physalis seeds, which cannot be identified to specific species, occur commonly in archeological sites in the southwestern USA (Kindscher et al., 2012). In fact, Physalis seeds have been found in ruins dating from as early as AD 298 at these archaeological sites in New Mexico: LA 109100 on Ceja Mesa west of Albuquerque (Dello-Russo, 1999); at the Basketmaker III/Pueblo I period (AD 650-900) site at River’s Edge west of the Rio Grande River; and north of Corrales (Brandt, 1991). Nutritional data on the native Physalis species is lacking, but tomatillos are anecdotally considered nutritious and these plants could provide an important source of additional vitamins and phytochemicals. Tomatillos contain alkaloids, as is common in the Solanaceae family (Whitson and Manos, 2005). Physalis species were used medicinally in the
past, and now are the subject of much research as our colleagues have discovered 14 new compounds in P. longifolia Nutt., including one which has strong and significantly unique anticancer properties (Zhang et al., 2011; Zhang et al., 2012). SUMMARY AND FUTURE DIRECTIONS North American athletes and other consumer groups use natural health products (NHP) to improve health and performance. Many NHP and dietary supplements are derived from indigenous/traditional knowledge systems. The central region of North American is largely comprised of one of the largest biomes in North America -the grasslands- as well as a variety of physiographic regions rich in ethnomedicine, past and present. For this reason, scientists in Canada and the USA are exploring the bioactive properties of plants from this region. Identified herein are several native and naturalized bioactive North American plants that could help improve the health and performance of athletes, and other consumers, when used as NHP, functional foods, and ergogenic aids. Bioactive plants and their compounds used for dietary supplements and NHP require further study regarding: identification, standardization, mechanism of action, toxicology. Future work should also elucidate plant compound benefits in human physiology, when consumed in a whole food matrix such as a functional food.
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Source of Support: Nil
Webster D, Lee TD, Moore J, Manning T, Kunimoto D, LeBlanc D, Johnson JA, Gray CA (2010). Antimycobacterial screening of traditional medicinal plants using the microplate resazurin assay. Can. J. Microbiol. 56:487–494. Whitson M, Manos PS (2005). Untangling physalis (Solanaceae) from the physaloids: A two-gene phylogeny of the physalinae. Syst. Bot. 30:216–230. Wink M (2010). Biochemistry of plant secondary metabolism. West Sussex: Blackwell Publishing. Zhang H, Samadi AK, Gallagher, RJ, Araya JJ, Tong X, Day VW, Cohen MS, Kindscher K, Gollapudi R, Timmermann BN (2011). Cytotoxic withanolide consituents of Physalis longifolia. J. Nat. Prod. 74:2532–2544. Zhang H, Motiwala H, Samadi AK, Day V, Aube J, Cohen MS, Kindscher K, Gollapudi R, Timmermann BN (2012). Minor withanolides of Physalis longifolia: Structure and cytotoxicity. Chem. Pharm. Bull. 60:1234–1239.
Conflict of Interest: None Declared
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 583–598 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Research article PREDICTION OF MOLECULAR TARGET OF SISSOTRIN USING DOCKING TECHNIQUES Zaman Aubhishek1, 2*, Shafrin Farhana2 1
Department of Genetic Engineering and Biotechnology, University of Dhaka, Dhaka-1000, Bangladesh. Molecular Biology Laboratory, Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh. *Corresponding author:E-mail: aubhishek@gmail.com; 2
Received: 14/09/2012; Revised: 31/10/2012; Accepted: 01/11/2012
ABSTRACT Sissotrin is a major small molecule found abundant in the herbal plant Dalbergia sisso (Sisso/Black Wood/Rose Wood). The plant is widely found and used in Bangladesh for medical purposes. However, how this small molecule mediates its action in the body is poorly understood. One of the applications of the plant is to cure bleeding disorders such as thrombocytopenic purpura. However what macromolecules inside the cell interact with the small molecule and finally mediate this action is yet to be confirmed. Here, in this study, it was predicted that sissotrin binds Platelet Derived Growth Factor Receptor- a molecular target hypothesized by docking technique- and thus initiates the subsequent drug action. This study confirmed the basis of the traditional herbal practice and can be useful further for developing an even more effective synthetic drug mimicking sissotrin structure in the future.
KEYWORDS: Dalbergia sisso, Platelet Derived Growth Factor Receptor, Sissotrin.
To Cite this article: Zaman A, Shafrin F (2012), PREDICTION OF MOLECULAR TARGET OF SISSOTRIN USING DOCKING TECHNIQUES, Global J Res. Med. Plants & Indigen. Med., Volume 1(11), 583–598
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 583–598
INTRODUCTION Dalbergia sisso (Figure 1) is a medium to large sized deciduous tree. Leaves of this plant are imparipinnate; leaflets range between 3–5 per leaf. Leaves are usually, 3–7 Cm long. They are distant from each other and alternate in nature. Flowers of the plant are yellow in axillary panicles and shorter than the leaves. Leaves contain sissotrin (Paulo, Martins et al., 2008) and an isoflavone-O-glycoside (Asif and Kumar 2011; Dixit, Chillara et al., 2012; Vasudeva and Vats 2011; Hajare, Chandra et al., 2001). Flowers contain biochenin A, tectorigenin, 7, 4-dimethyl tectorigenin and 7O-methyltectorigenin. Green pods contain meso-inisitol, 7-O-methyltectorigenin and its 4´-rhamnoglucoside. Mature pods contain isocaviumin, tectorigenin, dalbergin, biochanin A and 7-hydroxy-4-methyl coumarin, 7-Oglucosides of tectorigenin, caviunin and tannins. Stem bark contains dalberginone, dalbergin, methyldalbergin, 4-phenylchromene, dalbergichromene and isotectorigenin (Farag, Ahmed et al., 2001; Singh and Bhati 2003; Shrestha, Amano et al., 2008; Yadav, Yadav et
al., 2008; Jaiganesh, Akilandeshwari et al., 2009). Heartwood contains a range of different oils and small molecules- dalbergin, nordalbergenones, dalbergichromene 3, 5dihydorxy-trans-stilbene, biochanin A and a allylphenol of latifolin type - dalbergiphenol (Ghani, 2003; Rastogi & Mehrotra, 1993). Platelet-derived growth factor receptors (PDGFR) (Shim, Liu et al., 2010) are cell surface receptors with tyrosine kinase activity (Escobedo, Barr et al., 1988; Wang, Pennock et al., 2004). They belong to the members of the platelet derived growth factor (PDGF) family (Williams 1989; Mantur and Koper 2008). PDGF subunits (named –A and –B) are important regulators for cell proliferation and differentiation, cell growth, development and many diseases including cancer (Rocconi, Matthews et al., 2008). Two forms of the PDGF-R, alpha and beta are each encoded by a different gene. The bound growth factor type determines whether PDGFR homo or heterodimerizes (Faller, Mundschau et al., 1994).
FIGURE 1: Dalbergia sisso plant.
The plant is found abundantly in Chittagong Hill Tracts (CHT) and in the northern zone of Mymensingh district in Bangladesh. Although plants from both the location seems to be similar, slight variations in phenotype may well be subject to further investigation. In Bangladesh varieties of the plant, if any, is not well classified.
The principal function of platelets is to prevent bleeding (Vieira-de-Abreu, Campbell et al., 2012). If the number of platelets is too low, excessive bleeding can occur (Mahla, Hochtl et al., 2012; Podd 2012). However, if the number of platelets is too high, blood clots can form (thrombosis) (Centemero and Stadler
2010; Ten Cate 2012), which may obstruct blood vessels and result in events such as stroke, myocardial infarction, pulmonary embolism or the blockage of blood vessels to other parts of the body, such as the extremities of the arms or legs. Abnormality or disease of the platelets can be of wide ranges; Some of
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the examples include thrombocytopathy, thrombocytopenic purpura etc. (Parreira and Ducla-Soares 1969; Parreira and Ducla Soares 1969). Many plant extracts are assumed to cause the platelet aggregation and thereby reduce bleeding disorder (Ware and Suva 2011). The ultimate objective of the study was to identify a target molecule for sissotrin and hence propose a mechanism how the small molecule may mediate its action in the body. MATERIALS To find out molecular target for small molecules, for example sissotrin, can be tricky for many reasons- firstly if one does not know where to look for the target, he/she may end up looking at a pool of completely different set of proteins. The deposition in the databases for such target pool is huge and it is not even possible by computers to carry out a databasewide docking screen with maximum precision (Marshall 1987; Jackson 1995). The Immunology Database and Analysis Portal (ImmPort) provides advanced information technology support in the production, analysis, archiving, and exchange of scientific data. It serves as a long-term, sustainable archive of data generated by investigators funded through the NIAID/DAIT. For selected targets, docking experiment can be carried out by using offline suits such as AutoDock or online servers such as PatchDock. AutoDock is a suite of automated docking tools. It is designed to predict how small molecules, such as substrates or drug candidates, bind to a receptor of known 3D structure (Handoko, Ouyang et al., 2012; Sandeep, Nagasree et al., 2011; Trott and Olson 2010). Current distributions of AutoDock consist of two generations of software: AutoDock 4 and AutoDock Vina (Adinarayana and Devi 2011; Trott and Olson 2010). In AutoDock atomic affinity grids can be visualised. This can help, for example, to guide organic synthetic chemists design better binders (Tiwari et al.,, 2009). AutoDock Vina
does not require choosing atom types and precalculating grid maps for them. Instead, it calculates the grids internally, for the atom types that are needed, and it does this virtually instantly (Goodsell, Morris et al., 1996; Osterberg, Morris et al., 2002; Morris, Huey et al., 2008). For analyzing structure upon docking several molecules viewing programs were used. UCSF Chimera is a highly extensible program for interactive visualization and analysis of molecular structures and related data, including density maps, supramolecular assemblies, sequence alignments, docking results, trajectories, and conformational ensembles (Pettersen, Goddard et al., 2004; Goddard, Huang et al., 2005). High-quality images and animations can be generated. Chimera includes complete documentation and several tutorials, and can be downloaded free of charge for academic, government, nonprofit, and personal use (Couch, Hendrix et al., 2006). PatchDock (http://bioinfo3d.cs.tau.ac.il/PatchDock/) is a computational tool for determining protein ligand or protein protein interaction (Schneidman-Duhovny, Inbar et al., 2005). Its algorithm is based on object recognition and image segmentation techniques used in Computer Vision. (Mashiach, SchneidmanDuhovny et al., 2010) I-TASSER (http://zhanglab.ccmb.med.umich.edu/ITASSER/) (Roy, Kucukural et al., 2010; Zhang 2008) server is an Internet service for protein structure and function predictions. 3D models are built based on multiple-threading alignments by LOMETS and iterative TASSER assembly simulations; function insights are then derived by matching the predicted models with protein function databases. SignalP 4.0 server predicts the presence and location of signal peptide cleavage sites in amino acid sequences from different organisms: Gram-positive prokaryotes, Gramnegative prokaryotes, and eukaryotes (Bendtsen, Nielsen et al., 2004). The method incorporates a prediction of cleavage sites and
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a signal peptide/non-signal peptide prediction based on a combination of several artificial neural networks (Bendtsen, Nielsen et al., 2004).
of searching target molecule for sissotrin needed to be done from the previously performed phenotypic assays after sissotrin treatment.
To verify predicted protein structure ANOLEA (Atomic Non-Local Environment Assessment) is a server that performs energy calculations on a protein chain, evaluating the "Non- Local Environment" (NLE) of each heavy atom in the molecule (Melo, Devos et al., 1997). On the other hand, QMEAN, which stands for Qualitative Model Energy Analysis, is a composite scoring function describing the major geometrical aspects of protein structures (Benkert, Kunzli et al., 2009; Benkert, Tosatto et al., 2009). Five different structural descriptors are used in QMEAN. The local geometry is analyzed by a new kind of torsion angle potential over three consecutive amino acids. A secondary structure-specific distancedependent pairwise residue-level potential is used to assess long-range interactions (Benkert, Tosatto et al., 2008). PROCHECK checks the stereochemical quality of a protein structure, producing a number of PostScript plots analysing its overall and residue-by-residue geometry (Laskowski, Rullmannn et al., 1996). It includes PROCHECK-NMR for checking the quality of structures solved by NMR.
A wide range of literature was searched for possible function of the molecule (Asa, Murai et al., 2012; Edwards, van den Heuvel et al., 2011; Lei, Rheaume et al., 2010; Williams 1989). Sissotrin has been associated to function as an astringent. From this phenotype we could identify the candidate protein molecules with whom it interacts. The ImmPort database (https://immport.niaid.nih.gov/immportWeb/) DAIT (Boyce, Assa'ad 2012; Graham 2011; Davis 1969) was used for this purpose ImmPort database returned potential immune related protein targets for the small molecule.
Mobyle is a online collection which provides an access to different software elements, in order to allow users to perform bioinformatics analyses (Alland, Moreews et al., 2005; Neron, Menager et al., 2009). fpocket (http://fpocket.sourceforge.net/), a tool integrated in Mobyle portal, is a very fast open source protein pocket (cavity) detection algorithm based on Voronoi tessellation (Le Guilloux, Schmidtke et al., 2009). METHODS Prediction of the target molecule The target molecules were at first searched at STITCH 3.1 (Kuhn, Szklarczyk et al., 2012) database by searching for sissotrin interacting protein molecule. However, no results were found for the compound. Therefore the process
The Reactome database (Croft, O'Kelly et al., 2011) retrieved the mechanism how the selected target proteins may mediate their action. Among all of them, some of the proteins were shortlisted to be tentative or potential targets for binding sissotrin molecule. The results in the study have only been mentioned for the target molecule predicted finally and that is- PDGFR. PDGFR sequences were collected from National Center for Biotechnology Information (NCBI) Protein database. Protein 3D structure is the best indicator of the function as only this gives good account of how protein in native conformation behaves in vivo in biologically significant microenvironment. Probable 3D structures were generated using I-TASSER. Protein signal sequences were found using SignalP 4.0 server. Computational Docking The 3D Ligand structure was downloaded from online database ChemDB (http://cdb.ics.uci.edu/) and was subsequently docked to protein 3D structure of the macromolecule. Ligand and macromolecule was docked using two programs- AutoDock Vina (Download link: http://vina.scripps.edu/download.html) and online tool PATCHDOCK.
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Prediction analysis Prediction of the structures were confirmed by online tool QMEAN and PROCHECK. The best 3D structure was selected on the basis of Ramachandran plot and Model quality Zscore. The best macromolecule structure was selected for docking with the downloaded ligand molecule. The ligand bound protein structures provided by PATCHDOCK were viewed by RasWin and PyMol (Lill and Danielson 2011). The AutoDock results were analyzed using AutoDock tools -1.5.6rc3 and PyMol.
To predict targets from phenotype the ImmPort database was used. The database retrieved potential candidate proteins for the phenotypic characteristic of the small molecule. The small molecule works as an astringent and thus causes hemostasis. From ImmPort database the potential candidates for the mentioned activity were shortlisted. Platelet Activating Factor and Platelet Derived Growth Factor Receptor were deemed as one of the potential targets (FIGURE 2).
RESULTS
The mechanism of action of PDGFR was explored using Reactome database. The database is a collection of all the interaction and pathway networks. From the Reactome results, (FIGURE 3) PDGFR roles in platelet aggregation or hemostasis was confirmed. Also several downstream cellular signals were found related to it. This makes PDGFR even a more potent target as due to the cascade effect often cells multiply drug action by proteins that carries initial signal in a whole network.
Prediction of the target molecule To predict the target molecule a thorough web wide search for literature was carried out.
In final analysis it was assumed that sissotrin may act as agonist to the receptor and thereby cause hemostasis.
Protein pockets, candidates for ligand binding sites, were found by fpocket tool. The Pocket-Finder tool results were screened with similar results found from binding. Thus this step can be viewed as a confirmatory step for the binding results.
FIGURE 2: Target molecule prediction from ImmPort database.
The ImmPort database is a comprehensive compilation for identifying protein of interest from a known phenotype or cellular process. The data shows prediction of PDGFR and PAF as potential target molecule.
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FIGURE 3: Reactome database search results.
The results show that PDGFR resides in an important posit position to carry out platelet aggregation. gation. Platelet aggregation is enhanced when the herbal mix is introduced.
FIGURE 4: PDGFR 3D structure.
PDGFR structure as viewed by UCSF Chimera. On the left is the structure with marked secondary attribute. On the right the structure is viewed in plain ribbon model. B sheet region is on the cc-terminal terminal region.
FIGURE 5: PDGF dimerization and signal peptide in the structure. structure
(Left) The he structure was downloaded from PDB database. The structure is hydrogen added (red and orange molecules). (Right) SignalP results. SignalP 4.0 result shows that there is a cleavage site between number 32 and 33 residue.
nce was collected from PDGFR sequence NCBI Proteins database and Uniprot. The protein structure was then generated using online server I-Tasser. Tasser. The structures were viewed by multiple molecule viewer tools such
as UCSF Chimera (FIGURE FIGURE 4), Jmol, PyMol, RasWin etc. The structure was predominated by beta sheets for which templates were abundant. However there were significant amount of loops and short helices were
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dimeric structure we produced. The structure does not explain for the whole sequence. Signal sequences that are proteolytically cleaved after translation is also predicted by SignalP 4.0 (FIGURE 5).
accommodated in the structure. This occurred due to the lack of templates already deposited in the database. Future X-ray crystallography data would help this case for certain. However the beta sheet dominated C-terminal region was the actual reported region for binding carbohydrate derivative ligands (Palmer, Smaill et al., 1998). The C-terminal region is also important for signal transduction, Tyr857 phosphorylation is important in for signaling which is situated near C-terminal (Chiarugi, Cirri et al., 2002). This means that the structure was suitable for docking experiment despite some limitations.
Docking was performed using AutoDock and PatchDock. Among 5 different I-Tasser generated structures, model_1 (c-score: -2.42) and model_2 (c-score: -2.65) had the best binding scores for the ligand. Following are the binding scores for model_2 with Sissotrin-
The PDGF bound PDGFR structure was downloaded from Protein Data Bank (RCSB PDB). The PDB structure shows that PDGF (a protein ligand) crosslinks both the PDGFR units (FIGURE 5). However the ligand did not bind to the same place. Thus this structure was only useful for analyzing the quality of the
According to the AutoDock binding result model_2 bound PDGFR with better affinity than model_1 and both bound the protein in the same pocket (TABLE 1). From the FIGUREs (FIGURE 6 and 8) it was evident that in model_2 the ligand was more protruded in the pocket.
Computational Docking
TABLE 1: AutoDock results for model_2 and sissotrin Ligand model2_ Sissotrin
Binding Affinity −9 −8.9 −8.7
rmsd/ub 0 3.84 2.889
rmsd/lb 0 2.276 1.585
FIGURE 6: model_2 bound with Sissotrin.
(Top) model_2 pocket (zoomed in 20 angstrom) as viewed by Pymol Negative control no binding in the protein (Bottom left) model_2 binding pocket has been marked in red dotted circle (Bottom right) model_2 bound with negative control. The control molecule was taken in random from PubChem database and no binding was found in any of the 5 randomly retrieved molecules. This figure in particular is for fusaric acid as a ligand. Docking was done by AutoDock. Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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The binding occurred near the C C-termianl region of the protein which is important for signal transduction. ansduction. The binding residues responsible were- Leu962, Lys 967, Gly 968, Ala 990, Val804 etc. A helix structure balances the docking of the molecule (FIGURE FIGURE 7).
target et molecule. None of the negative controls shared the binding region of PDGF. Similar docking experiments were done for model 1 of I-Tasser. Tasser. Although the binding scores were lower than the ones for the model 2 (TABLE 2 and FIGURE 8).
Negative controls were used for randomly selected ligand molecules to bind against the FIGURE 7: Ligand bound to the predicted pocket.
That thee predicted pocket as balanced by the helix is viewed by PyMol. Key residues were viewed by this.
TABLE 2:: Autodock results for sissotrin bound to model1 Ligand model1 _Sissotr in
Binding Affinity −8.8 −8.6 −8.5 −8.1
rmsd/u b 0 10.103 10.279 19.943
rmsd/lb 0 2.409 4.432 13.793
FIGURE 8:: AutoDock results for sissotrin binding to model 1.
(Right) the structure is viewed in AutoDock, the target molecule is shown is wireframe mode. (Left) target molecule is shown as surface mode.
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TABLE 3: PATCHDOCK results for sissotrin bound to model1 (best solutions are enlisted) model 1
Score
Area
1 4
6292 5502
734.70 700.20
Global energy −45.00 −40.09
ACE
Repulsive VdW
−96.85 −81.46
−25.76 −23.16
FIGURE 9: PATCHDOCK results.
The PATCHDOCK docking region is similar to that of the found one by Autodock. The ligand is marked in red and the model 2 structure is marked in grey. (Right) The key residues interacting with the ligand is marked in colors- Cyan, Blue, Yellow and Green.
TABLE 4: PatchDock results for sissotrin binding to model 2 (only best solutions are enlisted) Model 2
Score
Area
1 8
5762 5422
701.30 735.30
PATCHDOCK online docking tool was used to reproduce the docking results (TABLE 3 and 4 and FIGURE 9 and 10). For model 2 the results were very reproducible with the first solution in PATCHDOCK being the best solution. Similar molecules were also found to affiliate binding in this case proving that the binding was in the same location. The target molecule was then looked for possible active pockets in it using fpocket online tool integrated within RPBS Mobyle Portal. In total 85 pockets were found. Among which pocket 12 was the one where our target binding was predicted by docking tools. Amino acid residues from 950–970 positions formed a pocket neear the ligand interacting region (FIGURE 10).
Global energy −47.54 −46.44
ACE
Repulsive VdW
−179.38 −107.01
−3.38 −6.69
Prediction analysis The structures of the protein generated were tested by PROCHECK computational tool. Ramachandran plots for the target molecule structure were not ideal, yet considering the lack of templates for the structure it was quite a stable one (FIGURE 11 Left). Although the side chain parameters (data not given) showed considerable deviation from an ideal values estimated by the tool, the main chain parameters fit the ideal values perfectly (FIGURE 11 Right). QMEAN online tool was used to analyze the structure’s similarity with the templates found in the database. The score for model 2 and model 1 (not given) both was quite off the
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grid from an ideal value (FIGURE 12) which was somewhat expected as the lack of templates for a structure significantly affect the
scores. The problem can only be solved once enough raw crystal structure data would be available.
FIGURE 10: fpocket results.
(Top) Fpocket predicted total 85 pockets among which in the twelfth one was a good fit for our predicted binding site. (Bottom) All the pockets were shown using multiple colors.
FIGURE 11: PROCHECK results.
(Left) Ramachandran plot and (Right) Main Chain Parameters for model1 predicted by I-Tasser.
DISCUSSION The original protein is 1106 amino acid long. However there is a cleavage present between number 32 and 33 residue. The structure had a great number of residues for which no known templates has been found. However the c-terminal region of PDGFR, which till now has not been much annotated
has been predicted with good efficiency. The ligand also was bound in that region. Sissotrin bound the target protein molecule PDGFR in the C terminal region. C terminal region is known to be associated to the signal transduction function. Thereby it is hypothesized here that the ligand binding to the protein triggers a signal transduction cascade
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which as a final outcome causes platelet aggregation. The structures of the target molecule as predicted by online servers were not very satisfactory. Nevertheless, the structure had
FIGURE 12: QMEAN results.
QMEAN results show that the predicted scores for model 2 (marked by red cross) was far off the ideal values.
stable secondary structures in the C-terminal region and N-terminal region, two regions responsible for most of the ligand binding. QMEAN results show that the structure is deviated from already predicted protein structures as template.
FIGURE 13: Distances between activation loop and the ligand binding
Ligand is shown in red and the activation loop (around ASP850) site is shown as blue. The distances between them are calculated using RasWin Pick Distance command. They are 35.51A° distant.
However the binding region proposed here has not been reported before. Although there is a presence of activation loop (CDD: 173623) (Marchler-Bauer, Lu et al., 2010) and the Tyr 857, important phosphorylation site, near 35.51A° no concrete proof of mechanism could have been drawn between the two sites directly (FIGURE 13) (Chiara, Goumans et al., 2004). The docking experiment was quite reproducible. Two programs namely AutoDock and PATCHDOCK have been used to predict ligand binding sites and their affinity and in both the cases similar results were found. Molecular docking often generates false positive results, the rule of thumb in these cases are to carry out an in-vitro assay. This study is not beyond this rule too. The study needs to be validated by experiments such as X-ray crystallography or Protein Array Chip
studies. Protein Array Chip study would give out conclusive information about binding between the small molecule and the receptor. In-silico drug designing has been the actual motivation behind this study. Although the conclusions came from this study may not be exactly ideal, this study does create a model for further such experimental designs in the future. It is hoped that the study would in some ways be helpful to promote scientific herbal practices and ethnobotanic studies to some degree. CONCLUSION Sissotrin has been found to be abundant in the herbal plant Dalbergia sisso. The study predicts a potential molecular target of sissotrin to be PDGFR. It predicts that by binding to PDGFR as an agonist it cures bleeding
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disorders such as thrombocytopenic purpura. If validated in vitro, the study can play an
important role in further drug designing in the future from the herbal plant mentioned.
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Source of Support: Nil
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Conflict of Interest: None Declared
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 599–611 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Research article PHYTOCHEMICAL ANALYSIS AND ANTIOXIDANT ACTIVITY OF THREE ETHNO MEDICINAL PLANTS OF PACHAIMALAIS, TIRUCHIRAPPALLI DISTRICT, TAMIL NADU. Rekha S1, Parvathi A2* 1, 2
Herbal Study Centre, Department of Botany, Holy Cross College (Autonomous), Tiruchirappalli - 620 002, Tamil Nadu, India. *Corresponding author: E-mail: parvathi.adapa@gmail.com
Received: 05/10/2012; Revised: 27/10/2012; Accepted: 31/10/2012
ABSTRACT Pachaimalai, an unique hill of Tiruchirappalli district is situated in the Southern parts of the Eastern Ghats in Tamil Nadu, India. It is endowed with rich medicinal flora. The Malayali Gounder Tribes are the inhabitants of the hills who inherited rich traditional knowledge about curing properties of the hill flora. The present work was focused on the evaluation of phytochemical constituents and antioxidant activity of the leaves and fruits of Naravelia zeylanica, (L.) DC., Cardiospermum canescens, Wall. and Mallotus philippinensis, Muell. Arg. that are used by these tribes to cure wounds, herpes, viral and bacterial diseases. The phytochemical analysis revealed the presence of various phytochemicals like alkaloids, amino acids, flavonoids, indoles, phenols, polyphenolases, saponins, steroids, tannins and triterpenoids. Further, the quantitative estimation of phytoconstituents also showed markedly high amount of flavonoids, phenols, saponins, tannins, amino acids and sugars. The antioxidant activity of methanolic extracts evaluated by Diphenyl picryl hydrazyl (DPPH) free radical scavenging activity revealed rich amount of antioxidant content in the selected plants. The paper deals with the significance of these three plants in traditional medicine with respect to their phytochemicals. Key words: Pachaimalais, Phytochemical analysis, antioxidant activity, Diphenyl picryl hydrazyl (DPPH) assay, Naravelia zeylanica, Cardiospermum canescens and Mallotus philippinensis.
To Cite this article: Rekha S, Parvathi A (2012), PHYTOCHEMICAL ANALYSIS AND ANTIOXIDANT ACTIVITY OF THREE ETHNO MEDICINAL PLANTS OF PACHAIMALAIS, TIRUCHIRAPPALLI DISTRICT, TAMIL NADU., Global J Res. Med. Plants & Indigen. Med., Volume 1(11), 599–611
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 599–611
INTRODUCTION Tamil Nadu historically has been an agricultural state of India cherishes rich ethnobotanical knowledge about medicinal plants since ancient times. The Siddha system of medicine of Tamil Nadu is one of the oldest systems of medicine in the world. The forests of Tamil Nadu harbour rich medicinal plant wealth which is used in traditional systems of medicine in India since ancient times. Pachamalai is an important unique hill in Tiruchirappalli district and situated in the southern parts of the Eastern Ghats in Tamil Nadu. The hills spread over two districts namely Salem and Tiruchirappalli (75 Km away from Tiruchirappalli city). Its elevation ranges from 400–1200 m above Mean SeaLevel and have a relatively moderate climate owing to its altitude and vegetation. The forest comprises of a tropical thorny, dry deciduous and moist deciduous type of vegetation (Champion, 1961). Total geographical area of hill is about 14,277 ha. In Thamizh (Tamil) language Pachai means green and the Pachaimalai hills are greener than the nearby hills. The hills are rich reservoir of plant diversity including valuable native, endemic and rare medicinal plants. Majority of them are shrubs (115), herbs (71), trees (70) and
climbers (20) (Matthew, 1971; Rajadurai et al., 2009; Umavathi and Parvathi, 2012). The inhabitants of the hills are the “Malayali Gounder Tribes” who possess an intimate knowledge of the plants of hills and depend on the plants for medicines to cure various ailments. Distinction of some plants as medicinal plants conveys an important association between these plants and a set of traditional knowledge on their use in medicinal preparations to treat people, livestock or plant diseases. The traditional knowledge on the application of plants for different medicinal uses that has evolved and maintained is largely determined by the locally available biodiversity (Geetha Rani, 2010; Rekha, 2012). There is a need to explore the potentiality of the medicinal plants of Pachaimalais that are rich in therapeutic potential. The local communities inherited rich traditional knowledge about curing properties of the hill flora. Due to the decline in the number of traditional healers, the indigenous knowledge on traditional medicines is slowly vanishing away. The developing countries mostly rely on traditional medicines that involve the use of different plant extracts or the bioactive constituents of the plants (Mayank Gangwar et al, 2011).
Fig 1 Distribution of plant habits in Pachamalais
20 shrubs 70
115
Herbs Trees Climbers
71
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Keeping this view in mind, the present investigation was carried out on three potential medicinal plants of Pachamalai hills namely Naravelia zeylanica (L.) DC., Cardiospermum canescens. Wall., and Mallotus philippinensis. Muell. Arg. for their phyto-chemical constituents and anti-oxidant activity. The hill plant Naravelia zeylanica, (L.) DC., belongs to the family Ranunculaceae. The genus Naravelia, DC., comprises of only one species. It is locally known as vathamkolli in Tamil language. The root and stem paste of Naravelia zeylanica is used to treat rheumatism, itches, scabies, allergies, headache and back pain (Arun Vijayan et al., 2007; Ramachandran et al., 2009). The leaf and stem of the plant are used traditionally to cure inflammation, skin diseases, arthritis, headache, wounds and ulcers (Ayyanar et al., 2005). Leaf of this plant was reported to be having antiulcer, anthelmintic, anti-inflammatory and wound healing activity (Ashoka Shenoy et al., 2009). The traditional medicine practioners use the juice of leaf and stem for treating intestinal worms, psoriasis and dermatitis (Harsha et al., 2003). Cardiospermum canescens, Wall., belongs to the family Sapindaceae. The genus Cardiospermum L., comprises of 2 species. It is locally known as periya mudakathan in Tamil language. The whole plant has different properties like diaphoretic, diuretic, emetic, emmenagogue, laxative, refrigerant, rubefacient and stomachic (Duke and Ayensu, 1985). It is used in the treatment of rheumatism, nervous diseases, stiffness of the limbs and snakebite. Leaf juice mixed with cumin is consumed to relieve pain in the joints and given at the time of delivery. The roots are diuretic and used in treating liver disorders and dysentery. The leaf juice is used for the treatment of asthma (Chopra et al., 1986). The leaves are rubefacient and are applied as a poultice in the treatment of rheumatism (William et al., 2005). Mallotus philippinensis. Muell. Arg., belongs to the family Euphorbiaceae. The genus Mallotus Lour., comprises of about 150 species in the world, of which 20 species have been reported from India
(Santapau et al., 1973) and 11 species with 2 varieties were reported from Tamil Nadu (Henry et al., 1987). The ethnomedicinal plant Mallotus philippinensis, (Lam.) Muell. Arg., locally known as sindhura manjal has been used medicinally for a long time throughout India (Maheshwari et al., 1980; Sadhale et al., 2004; Thakur et al., 2005). It is used in ayurvedic medicine in the name of Kampillaka to relieve cough, constipation, flatulence, wounds, ulcers, renal hemorrhages and poisonous affections. This plant is applied externally as a treatment for skin disorders such as scabies and cutaneous troubles, tinea, herpes and other parasitic infections. The leaves and bark are used in India as poultice to skin disorders and the pounded seeds are applied over the wounds (Wiart, 2006). MATERIALS AND METHODS The materials for the study were collected during October, 2011- February, 2012. The voucher specimens (501,602,703) were deposited at Herbal Study Centre, Department of Botany, Holy Cross College (Autonomous) Tiruchirappalli, India. The plants were taxonomically identified with the aid of “Flora of the Presidency of Madras” (Gamble, 1957) and “Flora of Tamil Nadu Carnatic” (Matthew, 1983). Extraction of plant material The collected parts (leaves and fruits) of selected plants were cleaned, shade dried and powdered in an electric grinder. Fifteen grams of powdered plant materials were soaked in 100 ml. of 80% ethanol and incubated for 24 h. using ethanolic extracts various tests were carried out (Gibbs, 1974; Harborne, 1984; Trease and Evans, 1989). The tests included alkaloids, amino acids, carbohydrates, flavonoids, indoles, phenols, polyphenolases, proteins, saponins, steroids, tannins and triterpenoids. Qualitative detection of Phytochemicals Qualitative phytochemical tests were performed on fresh and ethanolic extracts of leaves and fruits of the selected medicinal herbs
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for the identification of their bioactive components. The extracts were treated with various chemical reagents and their colour reactions were observed to identify the presence of phyto-constituents such as alkaloids, amino acids, steroids, flavonoids, polyphenolases, triterpenoids, proteins, carbohydrates, saponins and tannins. Quantitative detection of phytochemicals After the confirmation of presence of amino acids, proteins, carbohydrates, flavonoids, phenols, saponins and tannins by preliminary phytochemical analysis, the coarse powder of the plant material was taken up for the quantitative estimation. Determination of total protein content: The total protein content in extracts was determined (Lowry et al., 1951). Fifty milligrams of sample was homogenized with 5 ml of ice-cold phosphate buffer and centrifuged at 2000 rpm for 5 min. To the supernatant solution, equal volume of 10% icecold Tricholro acetic acid (TCA) was added and incubated for 10 min. at 4°C for an hour. The precipitated protein was centrifuged and the pellet was dissolved in one ml of 0.1N NaoH. 0.5 ml of the protein solution was mixed with 5 ml of alkaline copper reagent. It was shaken well and allowed to stand at room temperature for 10 min. Then, 0.5 ml of folin – ciocalteau reagent was added and the volume was made upto a known quantity using distilled water. Blank was prepared without the sample extract. After 30 min the optical density of the solution was read at 660 nm in Spectronic–20 D. Determination of total free amino acid: Hundred milligram of dried sample powder was homogenized with 5 ml of 80% ethanol and centrifuged at 2000 rpm for 10 min. The pellet was re-extracted with the same solvent and centrifuged again. The supernatant were pooled. To the supernatant equal volume of petroleum ether was added to remove the chlorophyll pigments using separation funnel. The lower layer was taken as sample. 0.5 ml of
acetate buffer was added to the 1 ml of alcoholic extract, followed by 1 ml of 1% ninhydrin. The reaction mixture was heated for 15 min. in a boiling water bath at 100ºC for colour development. It was then cooled and the volume was diluted to 10 ml with distilled water. For blank, 0.5 ml distilled water was taken and all the reagents were added and carried out as above. The colour intensity was measured at 570 nm (Moore and stein, 1948). Determination of total carbohydrates: Hundred milligram of dried sample powder was homogenized with 5 ml of 80 % ethanol and centrifuged at 2000 rpm for 10 min. The pellet was re-extracted with the same solvent and centrifuged again. The supernatant were pooled. To the supernatant, equal volume of petroleum ether was added to remove the chlorophyll pigments using separation funnel. The lower layer was taken as sample. 1 ml of protein free carbohydrate solution was mixed with 3 ml of the anthrone reagent (0.2% in Conc.H2SO4). The reaction mixture was heated for 5 min. in a boiling water bath at 100ºC with the marble on the top of the test tube to prevent loss of water by evaporation. Suitable reagent blank was prepared. The colour intensity was measured at 620 nm in a Spectronic -20 (Mahadevan and Sridhar, 1982). Determination of total Flavonoids Two gram of the plant sample was extracted repeatedly with 100 ml of 80% aqueous methanol at room temperature. The whole solution was filtered through whatman filter paper No. 42 (125 mm). The filtrate was later transferred into a crucible and evaporated to dryness over a water bath and weighed to a constant weight (Bohm and Kocipai Abyazan, 1974). Determination of total phenols: 100 mg of the sample was extracted with 5 ml of 80% ethyl alcohol and centrifuged at 2000 rpm. The supernatant was taken for assay. One ml of folin–ciocalteau reagent was added to 0.5 ml of the alcoholic extract of the sample. 2 ml of 20 % sodium carbonate was added and
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heated for one min. After cooling, the solution was made up to 10 ml with distilled water. A blank was prepared by adding all the reagents except the sample. The absorbency was read at 650 nm in Spectrophotometer (Mahadevan and Sridhar, 1982). Determination of Saponins The ground sample of 2 g of each was placed in a conical flask and 100 ml of 20 % aqueous ethanol were added. The samples were heated over a hot water bath for 4 h with continuous stirring at about 55ºC. The mixture was filtered and the residue was re-extracted with another 200 ml of 20% ethanol. The combined extracts were reduced to 40 ml over water bath at about 90°C. The concentrate was transferred in to a 250 ml separation funnel and 10 ml of dimethyl ether was added and shaken vigorously. The aqueous layer was recovered and the ether layer was discarded. The purification process was repeated. 10 ml of n– butanol was added to it. The combined n– butanol extracts were washed twice with 10 ml of 5% aqueous sodium chloride. The remaining solution was heated in a water bath. After evaporation, the samples were dried in the hot air oven to a constant weight and the saponin content was calculated (Obadoni and Ochuko, 2001). Determination of Tannin: 500 mg powdered sample material was transferred to 250 ml conical flask containing 75 ml of distilled water. The contents in the flask were boiled for 30 min, centrifuged for 2000 rpm for 20 min. The supernatant was collected in 100 ml volumetric flask and made up to a known volume. One ml of the sample extract was transferred to a 100 ml volumetric flask containing 75 ml of distilled water. To this, 5 ml of Folin-Denis reagent and 10 ml of sodium carbonate solution were added and diluted to 100 ml. It was shaken well and left for 30 min. and the absorbance was read at 700 nm against a reagent blank water (Sadasivam and Manickam, 1992).
Evaluation of antioxidant activity Extraction of plant materials: The powdered plant materials were extracted by maceration. About 2.5 g of the dried, pulverized materials were extracted in 10 ml of methanol at room temperature for 3 days. The mixture was filtered using Whatman No. 1 filter paper. The extracts of methanol were stored in air-tight glass bottles at room temperature. Phosphomolybdenum antioxidant assay The total antioxidant activity of the extract was evaluated by the phosphomolybdenum assay method Prieto et al., (1999). It is based on the reduction of Mo (VI) to Mo (V) by the extract and subsequent formation of a green phosphate-Mo (V) complex in acetic condition. 0.2 ml (100 mg) of extract was combined with 3 ml of reagent solution (0.6M sulfuric acid, 28 mM sodium phosphate, 4 mM sodium phosphate and 4 mM ammonium molybdate). The reaction mixture was incubated at 95°C for 90 min. Then, the absorbance of the solution was measured at 695 nm using a UV-visible spectrophotometer against a reagent blank after cooling to room temperature. The antioxidant activity was expressed as the number of grams equivalent of ascorbic acid. Free Radical Scavenging activity (DPPH Assay) Free radical scavenging potential of extracts was determined by 2, 2-diphenyl-1picrylhydrazyl (DPPH) assay by the method established by Brand-Williams et al., (1995). To 3 ml methanol solution of DPPH (20 µg/ml), 2 ml (100 mg/ml) of methanol extract of sample was added. The mixture was incubated in dark at room temperature for 30 min. The degree of free radical scavenging activity in presence of different samples and their absorbance were measured by using UV spectrophotometer at 517 nm. The degree of free radical scavenging activity was expressed as;
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steroids, tannins and triterpenoids are seen in abundance in the leaves and fruits of N. zeylanica and C. canescens. These results are in par with the reports of Lalitha Easwaran et al., (2011) and Thirupal Reddy et al., (2010) .The leaves of M. philippinensis showed the presence of amino acids, carbohydrates, cardiac glycosides, flavonoids, phenols, proteins, polyphenolases, steroids, tannins and terpenoids which are similar to previous report (Jayaraman Velanganni et al., 2011).
Scavenging Activity (%) = {(A control – A sample)/ (A control)} X 100 A control = Absorbance of DPPH alone, A sample = Absorbance of DPPH along with extracts. RESULTS AND DISCUSSION Preliminary phytochemical screening of selected plants is presented in the Table-1. From the table, it is noted that the presence of bioactive compounds like alkaloids, amino acids, flavonoids, polyphenolases, saponins,
QUALITATIVE ANALYSIS Table-1 Test with 80% ethanolic extracts S.No
1.
2.
3.
Parts used
AL
AA
CA
CG
FL
HCN
HW
IN
PH
PR
SA
ST
TA
TRIT
Leaves
+
+++
−
−
+++
−
+++
−
++
+++
+
+
+
++
Fruits
+
+++
−
*
+++
*
*
−
−
+++
*
+
+
+
Cardiospermum canescens ,Wall.,
Leaves
+
+++
+++
−
+++
−
−
−
−
+++
+
+
+
++
Fruits
+
+++
+
*
+++
*
*
+
+++
+++
*
++
++
−
Mallotus philippinensis, Muell.Arg.,
Leaves
−
+++
+++
+
+++
−
+++
−
+++
+++
+
+
++
+++
Name of the plants
Naravelia zeylanica,(L.) DC.
+++ = High intensive; ++ = medium intensive; + = low intensive; − = Negative; * = Not tested AL= Alkaloids; AA= Amino Acids; CA= Carbohydrates; CG = Cardiac glycoside; FL= Flavonoids; HW= Hot water test; IN= Indoles; PH= Phenols; PR= Proteins; SA= Saponins; ST= Steroids; TA= Tannins; TRIT= Triterpenoids
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QUANTITATIVE ANALYSIS Table Table-2 Estimation of primary metabolites
S.NO
Sugars mg/g dry tissue
Amino acids mg/g dry tissue
Proteins mg/g dry tissue
10.560
4.950
1.260
11.820
6.750
Leaves
10.920
6.750
0.550
Fruits
6.480
2.850
0.300
Leaves
10.800
5.550
1.320
Parts used
Name of the plants
Leaves Naravelia zeylanica, (L.) Dc.
1.
Fruits
Cardiospermum canescens ,Wall.,
2.
Mallotus philippinensis, Muell., Arg.,
3.
0.760
Fig-22 Quantitative estimation of primary metabolites 11.82 12
10.92
10.56
10.8
mg/g of dry tissue
10
8 6.75 6
6.75
6.48 5.55
4.95
Sugars Amino acids
4
2
2.85
Proteins 1.32
1.26 0.76
0.55
0.3
0 Leaves
Fruits
N.zeylanica
Leaves
Fruits
C.canescens
Leaves M.philippinensis
sample
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QUANTITATIVE ANALYSIS Table- 3 Estimation of secondary metabolites Phenols Flavonoids Saponins Parts mg/g Name of the plants mg/g dry mg/g dry used dry tissue tissue tissue
S.No
Leaves
26
4.515
17.747
58
Fruits
10
5.394
12.676
49
Leaves
51
3.196
6.227
Fruits
55
3.556
7.561
Leaves
47
6.713
21.128
Naravelia zeylanica, (L.) Dc. 1.
2.
3.
Cardiospermum canescens, Wall.,
Mallotus philippinensis, Muell.Arg.,
Tannins mg/g dry tissue
47
37
39
Fig 3 - Quantitative estimation of secondary metabolites 58
60
55 49
51 47
mg/g of dry tissue
50
47
30
39
37
40
26
flavonids 21.128
20
17.747
saponins
12.676 10 10
4.515
5.394
6.227 3.196
7.561 3.556
6.713
leaves
fruits
leaves
fruits
leaves
0
N.zeylanica
phenols
Cardiospermum canescens
Mallotus philippinensis
Sample
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tannins
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TABLE -4 Antioxidant content and Free radical scavenging activity S.No
1.
2.
3.
Name of the plants
Parts used
Total antioxidant content mg/g dry tissue
Free radical scavenging activity in percentage
Leaves
3.23064
80.1980
Fruits
3.20756
87.1287
Leaves
3.11526
78.271
Fruits
2.16914
88.118
Leaves
3.69216
84.1584
Naravelia zeylanica, (L.) Dc.
Cardiospermum canescens, Wall.,
Mallotus philippinensis, Muell. Arg.,
Fig 4- Antioxidant and Free radical scavenging activity 4
3.69216
mg/g of dry tissue
3.5 3.23064
3.20756
3.11526
3 2.5
2.16914
2 antioxidant acivity
1.5 1
Free radical scavenging activity
80.20%
87.13%
78.27%
88.12%
84.16%
Leaves
Fruits
Leaves
Fruits
Leaves
0.5 0
N.zeylanica
C.canescens M.philippinensis Sample
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The plants were also quantified for the metabolites such as amino acids, sugars, proteins, flavonoids, phenols, saponins and tannins. The results are presented in Tables 2 and 3 and Figures 2 and 3. From the tables, it is understood that the sugar content of the fruits of N. zeylanica was found to be highest (11.820 mg/g) and low amount in C. canescens (6.480 mg/g). Whereas, in the leaves of C. canescens, M. philippinensis and N. zeylanica the sugar content varied from 10.920 mg/g, 10.800 mg/g to 10.560 mg/g respectively. The previous report (Subasini et al., 2011) on C. halicacabum is on line with our observation. Similarly, the same amount of amino acids was registered in the fruits of N. zeylanica and leaves of C. canescens (6.750 mg/g) followed by the leaves of M. philippinensis (5.550 mg/g) and leaves of N. zeylanica (4.950 mg/g). The lowest amount of amino acids was registered in the fruits of C. canescens (2.806 mg/g).The total protein content ranged between the 0.300 mg/g (fruits of C. canescens) and 1.320 mg/g (leaves of M. philippinensis). The fruits of C. canescens showed the highest amount of flavonoid content (55 mg/g), whereas, the fruits of N. zeylanica showed the lowest amount (10 mg/g). Similarly, the leaves of C. canescens registered the high amount (51 mg/g) of flavonoid content followed by M. philippinensis (47 mg/g) and N. zeylanica (26 mg/g). Our results find supportive evidence from the previous reports (Suthar Singh et al., 2011 and Bimal Kumar Ghimire et al., 2011). The high amount of phenol content was registered in the leaves of M. philippinensis (6.713 mg/g) that followed by fruits of N. zeylanica (5.394 mg/g). The phenol content of leaves ranged between 3.196 mg/g (C. canescens) and 4.515 mg/g (N. zeylanica) as evidenced from the work of Bimal Kumar Ghimire et al., (2011). The leaves of M. philippinensis contained highest amount of saponins (21.128 mg/g) followed by leaves of N. zeylanica (17.747 mg/g). The lowest amount (6.227 mg/g) was registered in the leaves of C. canescens. On the contrary to our observation, Sutharsingh et al., (2011) in their study
reported the minimum amount of saponin. The fruits of N. zeylanica showed high amount (49 mg/g) of tannin followed by the fruits of C. canescens (37 mg/g). Whereas, the leaves of N. zeylanica registered high amount (58 mg/g) of tannin followed by C. canescens (47 mg/g) and M. philippinensis (37 mg/g). The results of antioxidant activity and free radical scavenging are presented in Table 4 and Figure 4. From the table, it is understood that the range of antioxidant content was highest in the leaves of M. philippinensis (3.69216 mg/g) followed by N. zeylanica (3.23064 mg/g) and C. canescens (3.11526g/g). Whereas the antioxidant content of fruits varied from 3.20756 mg/g (N. zeylanica) to 2.16914 mg/g (C. canescens). Our results find supportive evidence from the studies of Jagadeesan et al., (2011). Highest free radical scavenging activity was observed in the fruits of Cardiospermum canescens (88.118%) followed by Naravelia zeylanica (87.1287%). In addition, the leaves of three selected plants exhibited maximum free radical scavenging activity. CONCLUSION From our studies, it could be concluded that these three selected medicinal plants contained considerable amount of phytotherapeutants like alkaloids, flavonoids, saponins, terpenoids, tannins and strong antioxidant properties. In addition, all the three plants responded negatively for the Hydrogen cyanide (HCN). Therefore, it is suggested that these plant parts could be consumed without any hesitation in the form of various herbal preparations like decoctions, infusions, powders and tonics to cure varied ailments. The results of the study would provide the basis for further isolation and evaluation of major active principles of these plant materials to test their efficacy against various diseases. Further, the development of these phyto-chemicals as herbal drugs could play an important role in health care of developing countries.
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ACKNOWLEDGEMENTS The authors are grateful to UGC, New Delhi for providing research grant. And also thank
the authorities of Holy Cross College (Autonomous), Tiruchirappalli for providing necessary facilities to carry out the present research work.
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Matthew K M (1983). In: the flora of the Tamil Nadu Carnatic. 3:289. Publishers: the Diocesan press, Chennai. Mayank Gangwar., Dharmendra Kumar, Ragini Tilak., Tryambak D Singh., Sushil Kumar Singh., Goel RK., Gopal Nath (2011). Qualitative phytochemical characterization and antibacterial evaluation of glandular hairs covering of Mallotus phillippinensis fruit extract. Journal of Pharmacy Research. 4(11), 4214–4216. Moore S. and Stein W H (1948). Photometric ninhydrin methods for use in the chromatography of amino acids. J.Biol.Chem.176: 367–388. Obadoni B O. and Ochuko P O (2001). Phytochemical studies and comparative efficacy of the crude extracts of some Homostatic plants in Edo and delta States of Nigeria. Global J. Pure Appl.Sci.8: 203–208. Prieto P., Pineda M. and Aguilar M (1999). Spectrophotometric quantification of antioxidant capacity through the formation of a phosphomolydenum complex: Specific application to the determination of vitamin E. Analytical Biochem.269: 337–341. Rajadurai M., Vidhya V G., Ramya M. and Bhaskar A (2009). Ethno-medicinal plants used by the traditional healers of Pachamalai Hills, Tamil Nadu, India. Ethno-Med. 3 (1): 39–41. Ramachandran V S., Shijo Joseph and Aruna R (2009). Ethnobotanical Studies from Amaravathy Range of Indira Gandhi Wildlife Sanctuary, Western Ghats, Coimbatore District, Southern India. Ethnobotanical Leaflets. 13: 1069– 1087.
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Rekha S (2012). Phytochemical analysis and antioxidant activity of three ethno medicinal plants of Pachaimalais Tiruchirappalli district, Tamil Nadu. M.Phil., thesis. Sadasivam S. and Manickam A (1992). Biochemical methods for agricultural sciences.Wiley Eastern Ltd., Madras. PP.196–197. Sadhale N. and Nene Y L (2004). On Elephants in Manasollasa-2.Disease and treatment. Asian Agri.His. 8: 115–127. Santapau H. and Henry A N (1973). A Dictionary of Flowering Plants in India. New Delhi: Council of Scientific and Industrial research; PP.1–198. Subasini U., Sundaraganapathy R., Ananda Thangadurai S., Malathy R. and Victor Rajamanickam G (2011). Determination of nutritive value for certain South Indian indigenous species. Int. J. Pharm and Ind. Res. 1(1): 17–21. Sutharsingh R., Kavimani S., Jayakar B., Uvarani M. and Thangatirupathi A (2011). Antiinflammatory and anti-arthritic activities of aerial parts of Naravelia zeylanica (L.) DC. IJRPC .1 (3): 303– 307. Sutharsingh R., Kavimani S., Jayakar B., Uvarani M. and Thangatirupathi A (2011). Quantitative phytochemical estimation and antioxidant studies on aerial parts of Naravelia zeylanica DC. IJPSR. 2 (2): 52–56.
Source of Support: Nil
Thakur S C., Thakur S S., Chaube A K. and Singh S P (2005). An ethereal extract of Kamala (Mallotus philippensis, Muell,.Arg.) Lam. Seed induce adverse effects on reproductive parameters of female rats. Repro. Toxicol. 20 (1): 149–156. Thirupal Reddy B., Ali Moulali D., Anjaneyulu E., Ramgopal M., Hemanth Kumar K., Lokanatha O., Guruprasad M. and Balaji M(2010). Antimicrobial screening of the plant extracts of Cardiospermum halicacabum L. against selected microbes. Ethnobotanical Leaflets. 14:911 –919. Tistaert C., Dejaegher B., Chataigné G., Van Minh C., Quetin-Leclercq J. and Vander Heyden Y (2011).Dissimilar chromatographic systems to indicate and identify antioxidants from Mallotus species. Talanta. 83 (4): 1198–1208. Trease G E., Evans W C (1989).Text book of Pharmacognosy. 14th ed W.B. Sanders, London. Umavathi R., Parvathi A (2012). Sacred trees of temples of Tiruchirappalli, Tamil Nadu- The Natural and Ecological Heritage of India. GJRMI.1(6):225-233. William D., Warden C J H. and Hooper D (2005). “Pharmacographia India” A History of the principal Drug of Vegetable Origin. Srishti publication, New Delhi. PP: 366–367. Wiart C (2006), Medicinal plants of the Asia – pacific Drugs for the future. World scientific publishing Co.Pte.Ltd. Singapore.
Conflict of Interest: None Declared
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 612–619 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Review article NEED AND RELEVANCE OF FORMATION OF INDIAN SYSTEMS OF MEDICINE AND HOMOEOPATHY (ISM & H) POLICY 2002 IN INDIA Singh Balpreet1*, Kaur Rajvir2, Kumar Manoj3, Singh Amarjeet4 1
PhD Research Scholar, Centre for Public health, University Institute of Emerging Areas in Science and Technology, Panjab University, Chandigarh, India 2 Master in Public Health, Centre for Public health, University Institute of Emerging Areas in Science and Technology, Panjab University, Chandigarh, India 3 Assistant Professor, Centre for public health, University Institute of Emerging Areas in Science and Technology, Panjab University, Chandigarh, India 4 Professor, Department of Community Medicine, Post Graduate Institute of Medical Education and Research (PGIMER), Chandigarh, India *Corresponding Author: E-mail: drbalpreetsaini@gmail.com
Received: 19/09/2012; Revised: 25/10/2012; Accepted: 31/10/2012
ABSTRACT India has designed National Policy on Indian systems of Medicine and Homeopathy (ISM & H Policy) in 2002 to emphasis the development of Ayurveda, Siddha, Unani, Yoga, Naturopathy and Homoeopathy due to public patronage of these systems. Currently India has two health related policies, National Health Policy and ISM & H Policy. This study proposes to look into the need of formation of ISM & H Policy in 2002. An online questionnaire was designed to assess the views of Public Health experts and mailed to 100 experts. The tool was tested for content validity and a pilot study was done to find out the feasibility of the study. The data was analysed with the help of Microsoft Excel 2007 represented as percentages. Results showed that 61 out of 100 experts responded to questionnaire. 84% of respondents favoured formation of unified health policy. Most mentioned reason for formation of ISM & H policy separate from NHP was ‘difference in concepts of modern medicine and alternative medicine’ followed by ‘partiality with ISM & H’ and ‘for strengthening of ISM & H’. Respondents disfavouring the concept separate ISM & H policy opined that all health systems have common goal of health which can be dealt by single policy. Majority of respondents opined that there was need of ISM & H Policy separate from National health Policy as separate policy is essential for parallel growth of different health systems. KEY WORDS: ISM & H Policy, AYUSH, Public Health experts, Rationalization
To Cite this article: Singh Balpreet, Kaur Rajvir, Kumar Manoj, Singh Amarjeet (2012), NEED AND RELEVANCE OF FORMATION OF INDIAN SYSTEMS OF MEDICINE AND HOMOEOPATHY (ISM & H) POLICY 2002 IN INDIA, Global J Res. Med. Plants & Indigen. Med., Volume 1(11), 612–619
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INTRODUCTION Indian Systems of Medicine and Homeopathy (also known as AYUSH) enjoy significant public patronage in India [Unnikrishnan P, 2010]. Due to their strengths, these systems have sustained themselves from long past. Despite of public faith and strengths of systems, Indian systems of Medicine were dominated by Western Medicine during British rule. The denigration of Traditional wisdom reached its zenith in 1835, when Lord Macaulay settled the controversy over whether government should support indigenous or western learning by ordering that Western knowledge should be exclusively encouraged in all areas governed by East India Company. Thereafter only western medicine was recognized as legitimate and Eastern systems were actively discouraged [Macaulay TB, 1957; Wujastyk,D , 2008]. At the dawn of twentieth century, with the assertion of Indian nationalism, interest in Indian art and science reawakened and Indian systems of medicine began a gradual renaissance. Bhore Committee and Mudaliar Committee, which brought revolution in Health System of India, identified the importance of ISM. After that, the Indian government set up several high-level committees to advise it on the course of action it should adopt in relation to ISM [Bhore J, 1946; Mudaliar AL, 1962]. Chopra Report recommended the complete integration on ISM and Modern Medicine but due to the influence of the allopathic professionals on the design of health care systems, recommendations regarding the incorporation of indigenous practitioners into the national health services were simply not followed through [Chopra, 1948]. Despite of long years of planning process, ISM could not get the kind of financial and political support available to the dominant system of allopathy even though they have proven to be equally good and effective systems and are patronized regularly by millions of people in the country. Later on the strengths of traditional system was internationally recognized. The concern was voiced in the World Health Organization
(WHO) Assembly in early 1970s, debates that existing health services in countries like India were not meeting the needs of the majority, that some changes became possible. One was the adoption of the primary health care model which focused on developing community participation through the involvement of locally acceptable people like practitioners of traditional medicine [WHO, 1978]. The second was a joint UN Children’s Fund (UNICEF)/WHO study which also recommended the mobilization and training of indigenous practitioners (including traditional birth attendants or dais). In 1977, with the launch of the Community Health Worker Scheme, a fresh impetus was given to the attempt to involve rural, institutionally nonqualified, traditional practitioners as voluntary, paramedical community workers [Nichter M, 1980]. At the same time, serious efforts were also made to base primary health care strategies on the use of indigenous plant drugs grown in local herbal gardens (Unikrishnan, 2010). International apprehensions for Traditional Medicine, poor Health indicators and inaccessible modern medical services forced Government of India to work towards the development of Indian systems of medicine. As a result, National Health Policy formulated in 1983 assigned an important role to ISM in the delivery of primary health care. But in this policy ISM & H did not get their due position. Moving step forward separate department of Indian Systems of Medicine and Homoeopathy was created under Ministry of Health and Family Welfare, Government of India in 1995 (Unikrishnan, 2010). The most decisive step in development of ISM & H was formation of National Policy on Indian systems of Medicine and Homeopathy in 2002. Now India has two policies, National Health Policy 2002 and Indian Systems of Medicine and homoeopathy Policy 2002, which are directing the Nation towards better healthcare. But still goal of Health for All is a dream in our country. Keeping the above mentioned points in view, the present
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investigation was undertaken with the objective to assess need of formation of ISM & H Policy in 2002 separate from NHP. MATERIAL AND METHODS An online questionnaire was designed to assess the views of stakeholders on the need of a separate ISM & H policy in 2002. The tool was given to some of the experts for content validity. Based on their suggestions and recommendations, restructuring of tool was done. A pilot study was conducted on 20 AYUSH/Public Health experts attending 4th World Ayurvedic Congress to find out the feasibility of the study. Questionnaire was mailed to 100 stakeholders. Respondents included who were faculties of community medicine departments of medical colleges, Public health departments/schools, AYUSH colleges, personnel from various organizations (national, international like UNICEF, WHO and voluntary) working in the field of Public health, students of MPH, MD/MS throughout the country. The data was then analysed with the help of Microsoft Excel 2007. Frequencies, percentages, mean, median, mode were used to draw inferences as data set was most appropriate for it. The consent of the respondents was taken with the condition that all the information provided by him/her during the study will remain confidential. RESULTS Response Rate Online questionnaire was sent to 100 Public Health/AYUSH experts on their emails. 61 experts responded to the questionnaire. Therefore response rate was 61%. Composition of sample Sample was composed of researchers, consultants etc.
faculties, Faculty
respondents included Vice-Chancellor, Directors, Assistant Directors, Professors, Associate Professors, Assistant Professors of Public Health educational and research institutes. Scholars included MPH, MD/MS and PhD students. Researchers included scientists, research officers, research managers, principle investigators. International organizations included officials of WHO and UNICEF and associations means the members of associations like PHFI, EUREMA. Consultants revealed Public Health consultants. Others included chairman, coordinators, CEO of NGOs/Projects. Exploration of expert opinion on need of separate ISM & H Policy 50 experts considered that there was a need of a policy on ISM & H separate from National Health Policy while 11 experts denied the need of separate policy on ISM & H in 2002. (Figure 1) Forty Public Health experts elaborated reasons for need of a separate policy on ISM & H. According to one expert “The separate policy is particularly important for tackling and elucidating the public health issues in the AYUSH & LHT sphere.” Other expert expressed need of separate policy for a specific time stating that “The separate policy is needed till the system reaches at par with other well established scientific system.” Others emphasized on separate systems within ISM & H by saying that “AYUSH is not a single entity; it comprises of various systems each of which have a distinct identity. Therefore, a policy is needed that can have strategies for the growth of these systems in an environment of mutual accommodation.” One expert suggested separate policies for all systems within ISM & H “There should be separate policies for Homeopathy, Ayurveda and Unani, Siddha, because every pathy is different and cannot be treated alike.”
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Figure 1: Response to question ‘Was there a need of separate Policy on ISM & H?’
No 16%
Yes 84%
Reasons for need of separate policy on ISM &H Experts elaborated many reasons for existent separate policy on ISM & H (Figure 2) ISM & H as different systems Eleven experts considered that ISM & H are different systems with different concepts than modern system of medicine. So there was a need for a separate policy to address relevant issues. es. Various experts stated it as under “Indian System of Medicine derives strength from ancient wisdom. These are distinct from western medicine which emphasizes on illness rather than wellness. Indian system is holistic and natural.” “While a consumeristic approach drives modern medicine, ISM & H requires a holistic approach. There needs to be a different framework to drive ISM & H.” “Concepts and line of treatment in ISM is entirely different. Implementation requires special policy and policy makers must be expert in ISM itself.”
“… Because the case with ISM & H is different one cannot bring it on a common platform of research, drug policy, medical policy.” “It is our long traditional medical therapy has its own particulars ulars regarding treatment & Diagnosis.” “The perspective (of ISM & H) H is distinct from the western bio-medically medically oriented outlook of the dominant public health view.” “These sciences have something unique and need unique way.” Partiality with ISM & H Nine experts quoted partiality with ISM & H in combined health policy as a reason for separate policy on ISM & H. H Experts opined it as under “There was no policy research centres exclusively set up for ISM sector. Because of this many a time’s policies are formulated without taking in to consideration the requirement of this sector.” “ISM & H component is appearing in other policies since very long like in National Health policy 1983. A separate policy was essential to
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address multiple areas and issues specific to Indian traditional medicines.” “ISM & H was dying in many parts of India. Modern medicine was gaining head. A doctor to a common man most of the times meant a graduate in MBBS. The health system in India till recent times posted only doctors of modern medicine in the various health systems. Hence it was necessary to have a separate policy for ISM & H.” “The potential of ISM & H is not tapped fully.”
“Obviously there was need for separate policy. In majority of the states ISM & H is under the control of Allopathic fraternity.” “The reason is simple... just think on the result and you will realize that there is something going absolutely wrong.” “Over the years due to prevalence and popularity of allopath system of medicine, the tradition system did not get proper attention. Our health system is also a contributing factor for sidelining the traditional system. Separate policy will definitely help to get its place in Indian health system.”
Figure 2: Reasons for separate policy on ISM & H
ISM&H are different systems
5% 5% 27% 20%
Partiality with ISM&H Strengthening of ISM&H systems For accurate strategies and focused attention For awareness regarding ISM&H
20%
23%
Strengthening of ISM & H systems Eight experts considered separate ISM & H policy necessary for development of these systems. They explained it as under “To enhance d strengths and for standardization of these systems of medicine a separate protocol as well as policy is surely needed........we can’t focus until a separate body is formed to deal with d root problems of these systems of medicines. For laying down of scientific authenticity of our systems of
Other reasons
medicine...research and development is the key and can be achieved only with unified efforts and planning for which separate policy is must.” “There was an urgent need to protect, popularize, and preserve the Indigenous System of medicine from the onslaught of Allopathic System.” “No research efforts are made to employ the modern tools to verify and refine the concept
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related to diagnosis, preventive and treatment aspects (in previous combined policies).”
“To address sector vide approach” For awareness regarding ISM & H
“There was a need to leverage its strengths to improve the health indicators of our country. It is therefore necessary to make an inclusive agenda for health. Therefore, to prime the various AYUSH streams, it was necessary to frame a separate policy, nurture these streams and mainstream them into the health policy.” “To harness the strength and revival of ISM& H and to establish it into a system of medicine so that it is accepted by one and all a separate policy was required.” “Modern medicine is established so strongly in India that it is absolutely vital to give strength to ISM by a separate policy.” “Despite the long history and existence of Indian Systems of Medicine and Homeopathy, their potential and scope have not been fully realized and utilized by both the Central and State Governments. Therefore, a separate policy on ISM and H was long overdue.” For accurate strategies and focused attention Eight experts expressed the need of separate policy for accurate strategies and focused attention for development of ISM & H. Experts expressed it as under “A policy provides requisite impetus to the necessary strategies related to the issue concerned. So, a separate policy will definitely help.” “Policy frame work will provide a road map to action plan and modalities for periodic evaluation and self regulation.” “To provide focused attention for promotion and development of AYUSH systems.” To have a dedicated and focused approach
Three experts considered separate policy for public awareness regarding strengths of ISM & H. Other reasons One expert considered need of separate policy on ISM & H to develop system alternative to Allopathy by stating that “Search of alternative to dangerous allopathic medicines indicated need to revive Ayurveda and others Indian systems of medicine. Thus need of Separate policy.” One expert felt need of separate policy to abolish title of alternative medicine used for ISM & H by stating that “It's called Indian Systems of Medicine but still taken as alternative medicine. So, it needs separate policy that can give it the stand to be the main not alternative.” Reasons for disfavoring separate policy on ISM & H Ten experts stated that there was not any need of separate policy on ISM & H. four experts gave reasons in support of their answer. One expert considered no need of separate policy as goal of both policies is health by stating that “Both of them are concerned with the curing of diseases, maintenance of health and promotion of health of human population. Even though the way is different, the aim is same. So there is no need for a separate policy”. One expert rejected need of separate policy to modernize concepts of ISM saying that “Certain concepts in traditional medicines are really outdated. New concepts have to be introduced to make it relevant scientifically in today's context. So called modern medicine is nothing but a modern refined product of Traditional Medicine.”
“Indian system of medicine should be given due emphasis and importance.”
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One expert consider separate policy as hindrance to sustainable development to health system stating that “The strengths of these systems can be identified and highlighted with that of the Allopathic system and has to be popularized in a sustainable way.” And other expert consider separate policy as hindrance to mainstreaming of ISM & H DISCUSSION Srinivasav (1995) in his article entitled ‘National health policy for traditional medicine in India’ advocated the need of a separate policy on ISM & H [Srinivasan P, 1995]. He recognized four major effects of ISM Policy viz. to encourage researchers and practitioners to be more active and effective in their field; to check the proliferation of spurious practitioners who damage the credibility of indigenous system; to coordinate integrate and mutual acceptance of both health systems; encourage more allopathic doctors to prescribe simple and effective home remedies. He considered Sri Lanka’s comprehensive national health policy, which addresses & utilizes ISM appropriately, responsible for remarkable high standards of healthcare. He believed that same kind of policy in India would have tremendously beneficial effects on health status of nation. Findings of our study also support their views of ISM & H Policy as majority of Public Health/AYUSH experts favored need of a separate policy on ISM & H. They favored separate policy because of difference in concepts and issues of these systems. An expert elaborated it as “While a consumeristic approach drives modern medicine, ISM & H requires a holistic approach. There needs to be a different framework to drive ISM & H.” and other stated as “… Because the case with ISM & H is different one cannot bring it on a common platform of research, drug policy, and medical policy.” Thus a common course of action cannot be applied to traditional systems and modern one. Partiality with ISM & H also emerged as a major factor to support the separate policy on ISM & H. Although ISM &
H got place in NHP 1983, but this was not rightful. NHP 1983 could not direct the specific steps for development & utilization of ISM & H and focused on modern medical system alone. Ultimately NHP 1983 was failed to revive ISM & H. Experts realized that a separate policy was needed to frame specific strategies and focused approach. One expert felt need of separate policy to abolish title of ‘alternative medicine’ used for ISM & H as ISM were the only available health systems before British rule in India. These are still major health systems utilized by majority of rural Indian population. In Indian scenario, Allopathy, which entered few centuries ago and still limited to urban areas, is actually alternative to ISM. Even an expert from this study went a step ahead and promoted need of different policies for all health systems included in ISM & H. Banerjee (2002) believed that ISM & H Policy seems to strike some kind of balance for the first time, does this by exploring the possibilities of integration between the different systems of medicine [Banerjee M, 2002]. She considered that ISM & H Policy clearly takes the position of promoting medical pluralism and introduce strategies to mainstream the indigenous systems of medicine. She found that this policy was a first in recognizing, identifying different' stakeholders which implies that the role of the civil society, i.e., both the companies manufacturing medicines and the non-governmental organizations engaged in issues around Ayurveda, is seen to be important to include and involve in any government initiative. CONCLUSION Majority of Public Health/AYUSH experts in this study favored need of a separate policy on ISM & H because of difference in concepts and issues of these systems. Partiality with ISM & H also emerged as a major factor to support the separate policy on ISM & H. An expert even went a step ahead and advocated the need of different policies for all health systems included in ISM & H.
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 612–619
REFERENCES Banerjee M (2002). Public Policy and Ayurveda: Modernising a Great Tradition. Economic and Political Weekly, 37(12):1136–1146. Bhore J (1946). Report of the health survey and development committee. Technical report, Government of India, Delhi. Chopra RN (1948) Report of the committee on indigenous systems of medicine. Technical report, Ministry of Health, Government of India, New Delhi. “The Chopra Report”. Vol.1: Report and Recommendations. Macaulay T B (1957). Minute of 2 february 1835 on indian education.In: Young, G. M. (ed.) Macaulay, Prose and Poetry. Cambridge MA: Harvard University Press, 721–24. Mudaliar AL (1962). Report of the health survey and planning committee. Technical report, Government of India, Ministry of Health, Delhi.
Source of Support: Nil
Nichter M. (1980) Community Health Worker Scheme: A Plan for Democratisation Economic and Political Weekly, 15(1): 37– 43.
Srinivasan P (1995). National health policy for traditional medicine in India. World Health Forum, 16(2): 190–3. Unnikrishnan P, (2010) Role of Traditional Medicine in Primary Health Care: An Overview of Perspectives and Challenges. Yokohama International Social Science Research. 14(6): 58 (724)–77 (743). WHO
(1978). Declaration of Alma-Ata. International conference on primary health care, Alma-Ata, USSR, 6–12 September 1978. Technical report, 34 World Health Organisation.
Wujastyk,D. (2008). “The Evolution of Indian Government Policy on Ayurvedain the Twentieth Century,” chapter 3 in Dagmar Wujastyk and Frederick M. Smith (eds.), Modern and Global Ayurveda: Pluralism and Paradigms.New York: SUNY Press, pp. 43–76. ISBN: 9780791474907
Conflict of Interest: None Declared
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 620–628 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Research article PHARMACOLOGICAL STUDY TO ASSESS THE VIPAKA OF CERTAIN SAMANA & VICITRA PRATYAYARABDHA DRUGS IN ALBINO RATS Jadoun Anuruchi1*, Solanki S K2, Ashok B K3, Dwivedi R R4 1
PG Scholar, Dept of Basic Principles, IPGT & RA, Jamnagar, Gujarat, India PG Scholar, Department of Shalya Tantra, IPGT & RA, Jamnagar, Gujarat, India 3 Research Assistant, Pharmacology laboratory. IPGT & RA, Jamnagar, Gujarat, India 4 Professor and Head, Dept of Basic Principles. IPGT & RA, Jamnagar, Gujarat, India *Corresponding Author: E-mail: dranuruchi@gmail.com; Mob: +919377545238 2
Received: 14/09/2012; Revised: 31/10/2012 Accepted: 01/11/2012
ABSTRACT According to Ayurveda, all the drugs show three types of Vipaka (post digestive effect) after the digestion, which directly depend on their Rasas (tastes). Relation of Rasas (tastes) of substances with their Vipaka (post digestive effect) is given based on the concept of Samana and Vicitra Pratyayarabdhata. The present animal experiment deals with the concept of Vipaka (post digestive effect), and its effect as a part of applied aspect of concept of Samana and Vicitra Pratyayarabdhata on Koshtha related parameters. Nimba Patra and Bhumyamalaki which are having Samana and Pratyayarabdha properties respectively are selected for evaluation of Koshtha related parameters in albino rats. Wistar strain albino rats of either sex were divided in to three groups and test drugs were administered in the dose of 540 mg/kg. Parameters like body weight, food consumption, water intake, fresh and dry faecal output and food conversion ratio were assessed. Data generated was statistically determined by Student’s t test for paired and unpaired data. Administration of Nimba showed baddhavitkata (difficulty in excretion) by decreasing water intake, faecal output, faecal water content and number of faecal pellets, while opposite observations are made in Vicitra Pratyayarabdha drug Bhumyamalaki administered group which are per their Vipakas (post digestive effects). Thus this study provides unequivocal scientific basis to the concept of Arabdata of Ayurveda. KEY WORDS: Arabdhata, Samana Pratyayarabdha, Vicitra Pratyayarabdha, Koshtha, Nimba Patra, Bhumyamalaki.
To Cite this article: Jadoun Anuruchi, Solanki S K, Ashok B K, Dwivedi R R (2012), PHARMACOLOGICAL STUDY TO ASSESS THE VIPAKA OF CERTAIN SAMANA & VICITRA PRATYAYARABDHA DRUGS IN ALBINO RATS , Global J Res. Med. Plants & Indigen. Med., Volume 1(11), 620–628
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 620–628
INTRODUCTION: Ayurveda Samhitas presented several Ayurvedic concepts in concise form; hence they are difficult to understand. Therefore it is essential to explore and evaluate these concepts for their practical applicability by clinical and animal experimentations. It is easy to go through animal experiment before performing clinical trials because one can control a lot of conditions in animal experiment, which are not possible with clinical study. According to Ayurveda, all the drugs shows three types of Vipaka (post digestive effect) after the digestion, which directly depend on their Rasas (taste) (Acharya Yadavaji T, 2009a). Relation of Rasas (taste) of substances with their Vipaka (post digestive effect) is given based on the concept of Samana and Vicitra Pratyayarabdhata (Paradakara Hari S S, 2010a). Arabdhata means the origin of any substance by its unique conjugation and configuration of Panchamahabhutas (pentaelements) (Bhattacharya Shri Taranatha T, 2002), which determines all the properties of substances. Changes in the Arabdhata during digestion and metabolism are known as Pratyayarabdhata (Paradakara Hari S S, 2010a), which happens due to change in conjugation and configuration of Mahabhutas (penta-elements) during digestion and metabolism. When any substance is ingested, it is digested and metabolized by the action of different Agni (digestive power) i.e. Jatharagni, Bhutagni and Dhatwagni. During this whole process it decomposes and resynthesizes several times in form of breakdown and reformation of bonds between Panchamahabhutas (penta-elements) (Dhyani S.C., 2008). So, the Panchabhautika (pentaelementeric) composition of substance changes again and again, resulting in manifestation of Vipaka (post digestive effect), Veerya (potency), Prabhava (cause for specific action) etc. at different levels of Agni (digestive power). On the basis of results of these breakdown and re-synthesis processes, all the substances can be categorized in two categories. The substances whose
Panchabhautika (pentaelementeric) composition after re-synthesis remains same as that of substance in each instance, the substance is known as Samana Pratyayarabdha. In such type of substances the Vipaka (post digestive effect), Veerya (potency), Karma (action) of substance found in conformity with Rasa (taste), so determination of all the properties and action of substance is possible only by knowing its Rasa (taste) (Paradakara Hari S S 2010b). While the substances whose Panchabhautika (pentaelementeric) composition changes at either level of digestion once or more time and becomes different from the original substance, are known as Vicitra Pratyayarabdha. In such type of substances Vipaka (post digestive effect), Veerya (potency) and Karma (action) etc. of substances are not found in conformity with Rasa (taste), so determination of all the properties of substance is not possible only by knowing its Rasa (taste) and it becomes mandatory to know about every property of substance before using it (Acharya Yadavaji T, 2009b). Hence substances having same Rasa (taste) may also show different properties and action depending on their Vipaka (post digestive effect), Veerya (potency) etc. The present animal experiment deals with the concept of Vipaka (post digestive effect) and its effect as a part of applied aspect of concept of Samana and Vicitra Pratyayarabdhata. This will help to find out differences between Samana and Vicitra Pratyayarabdha substances at the level of Vipaka (post digestive effect). It is well known that Vipaka (post digestive effect) shows its effect on Dosha (body humours), Dhatu (tissues) and Malas (excretory products) (Acharya Yadavaji T, 2009c) & it is easy to assess the effect of Vipaka (post digestive effect) on Malas (excretory products), hence the present animal experimental model was designed to ascertain the effect of selected Samana and Vicitra Pratyayarabdha drugs on Koshtha related parameters in albino rats.
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MATERIALS AND METHODS:
dose of (1 ml/100 g) at morning hour (9– 10 AM).
Test drugs: Experimental protocol: The authenticated raw drugs namely Nimba Patra (Leaves of Azadirachta indica A. Juss) and Panchanga of Bhumyamalaki (Whole plants of Phyllanthus fraternus Linn) were collected from the pharmacy of Gujarat Ayurved University, Jamnagar. Both the drugs were shade dried and powdered, tablets of both drugs were prepared by adding 3% Gum acacia as per standard procedure. Among these drugs, Nimba Patra is having Samana Pratyayarabdha properties (i.e. Tikta rasa, Shita virya & Katu vipaka) and Bhumyamalaki is having Vicitra Pratyayarabdha properties (i.e. Tikta rasa, Shita virya & Madhura vipaka). The difference in the properties of drugs is mainly at the level of vipaka (post digestive effect). Animals: Eighteen healthy, young (6–7 weeks old), nulliparous, non-pregnant Wistar strain albino rats of either sex weighing between 170 ± 20 g were selected from animal house attached to the Pharmacology Laboratory, I.P.G.T. & R.A., G.A.U., Jamnagar. Animals were maintained on Amrut brand pellets obtained from Pranava Agro Ltd. and drinking water and exposed to ambient temperature, humidity and natural day and night cycles in individual metabolic cages. The experimental protocol was submitted to the animal ethics committee of the institute, and approval was obtained for conducting the experiment (Approval number - IAEC/9/11/19). Dose fixation and schedule: Dose of drugs (Powders prepared by crushing tablets) was fixed by extrapolating the human dose to rats based on body surface area ratio by referring to the table of Paget’s and Barnes (1964) (Laurence D.R. and Bacharach A.L 1964). On this basis, the rat dose was found to be 540 mg/kg. Drugs were suspended in deionized water at suitable concentration to prepare stock solution, freshly just prior to administration and administered orally in the
The study was carried out based on previous study which was designed by Dixit U D et al., (1995) to assess the effect of test drug on status of Agni (digestive power) (Dixit U.D. et al., 1995). The selected animals were divided in to three groups of six animals consists of three males and three females. The study was carried out in two phases, namely preliminary study and therapeutic study. Preliminary study of three days was carried out prior to the therapeutic study to understand and obtain base line data about the normal quantity of the parameters like body weight, food consumption, water intake, fresh and dry faecal output and food conversion ratio. In this particular phase, drug was not administered. Initial weight of each rat was recorded and they were placed in separate metabolic cages. All the parameters mentioned above were measured and recorded on routine basis. Statistical analysis: Data have been presented as Mean ± SEM. Difference in the groups was statistically determined by student’s t for both paired and unpaired data. Level of significance was set at P < 0.05. RESULTS: A uniform and normal progressive increase in body weight was occurred in control group. The similar type of weight gain was observed in Bhumyamalaki group, while in Nimba group comparatively less gain in body weight was observed in comparison to control group. However the changes were statistically nonsignificant (Table - 1). A marginal decrease in food consumption was observed in control group when the values of experimental phase were compared with values of preliminary phase. Administration of Nimba did not affect the food intake, while the
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Vicitra Pratyayarabdha drug Bhumyamalaki leads to 13.06% increase in food intake. However all the observations were found to be statistically non-significant. When the values were compared with control group,
Bhumyamalaki administered group shows apparent increase in food consumption, while Nimba shows marginal decrease in food consumption (Table - 2).
Table: 1 -- Effect on body weight Groups
Preliminary phase (g)
Experimental phase (g)
Actual change (g)
% change in comparison to Preliminary phase ### 183.28 ± 8.16 200.00 ± 10.27 16.73 ± 2.39 09.12 ↑ Control 185.16 ± 7.07 192.96 ± 6.78 7.79 ± 3.42 04.06 ↑ Nimba 194.80 ± 16.47 17.58 ± 9.18 09.91 ↑ Bhumyamalaki 177.22 ± 8.48 Data: Mean ± SEM ###P < 0.001 (Paired t test) ↑- Increase Table: 2 – Effect on food consumption
Groups Control Nimba Bhumyamalaki
Food consumption (g/100g) Preliminary Experimental Actual % change in comparison to phase phase change Preliminary phase 9.62 ± 0.41 8.98 ± 0.76 - 0.64 ± 6.65 ↓ 0.50 8.38 ± 0.85 8.42 ± 0.76 0.03 ± 0.01 00.47 ↑ 7.81 ± 0.95 8.83 ± 0.51 1.02 ± 1.00 13.06 ↑ Data: Mean ± SEM ↓- Decrease ↑- Increase Table: 3 -- Effect on water intake
Group
Control Nimba Bhumyamalaki
Preliminary phase
Water intake (ml/100g) Experimental Actual change phase
8.12 ± 1.09 8.26 ± 1.06 0.14 ± 1.98 9.12 ± 1.52 7.18 ± 1.29 2.03 ± 1.61 5.03 ± 1.59 6.97 ± 1.53 1.94 ± 1.04 Data: Mean ± SEM *P < 0.05 ↓- Decrease ↑- Increase
% change in comparison to Preliminary phase 01.72 ↑ 21.27 ↓ 38.57 ↑
Table: 4 -- Effect on weight of fresh faecal output
Group
Control Nimba Bhumyamalaki
Fresh faecal output (g/100g) Preliminary Experimental Actual change phase phase 2.35 ± 0.29 2.26 ± 0.45 1.73 ± 0.57
2.51 ± 0.30 0.15 ± 0.14 2.30 ± 0.46 0.04 ± 0.02 2.53 ± 0.65 0.79 ± 0.41 Data: Mean ± SEM ↑- Increase
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% change in comparison to Preliminary phase 06.81 ↑ 01.77 ↑ 46.24 ↑
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Table: 5 -- Effect on dried faecal output
Group Control Nimba Bhumyamalaki
Preliminary phase 1.51 ± 0.14 1.42 ± 0.18 1.17 ± 0.20
Dried faecal output (g/100g) Experimental Actual phase change 1.58 ± 0.09 0.07 ± 0.19 1.45 ± 0.22 0.03 ± 0.24 1.47 ± 0.25 0.30 ± 0.13 Data: Mean ± SEM ↑- Increase
% change in comparison to Preliminary phase 04.63 ↑ 02.11 ↑ 25.64 ↑
Table: 6 -- Effect on faecal water content Group Control Nimba Bhumyamalaki
Fresh faecal output Dried faecal output (g/100g) (g/100g) 0.15 ± 0.14 0.07 ± 0.19 0.04 ± 0.02 0.03 ± 0.24 0.79 ± 0.41 0.30 ± 0.13 Data: Mean ± SEM ↓- Decrease
% change 53.33 ↓ 25.00 ↓ 62.02 ↓
Table: 7 -- Effect on number of faecal pellets
Group
Control Nimba Bhumyamalaki
Preliminary phase
Number of faecal pellets Experimental phase Actual change
17.26 ± 2.00 17.09 ± 0.85 0.16 ± 1.77 17.68 ± 1.73 16.29 ± 2.60 1.39 ± 1.90 16.33 ± 1.97 17.31 ± 1.92 0.96 ± 1.12 Data: Mean ± SEM ↓- Decrease ↑- Increase
% change in comparison to Preliminary phase 0.98 ↓ 7.86 ↓ 6.00 ↑
Table : 8 -- Effect on food conversion ratio
Group
Control Nimba Bhumyamalaki
Food conversion ratio (g/100g) Preliminary Experimental phase Actual change phase 5.05 ± 0.37 4.37 ± 0.83 0.67 ± 1.03 5.03 ± 0.56 4.28 ± 0.54 0.75 ± 0.99 6.36 ± 1.12 5.44 ± 1.77 0.92 ± 1.46 Data: Mean ± SEM ↓- Decrease ↑- Increase
Water intake was marginally increased in control group when the values of experimental phase were compared with values of preliminary phase. Samana Pratyayarabdha drug Nimba shows 21.27% decrease in water intake in comparison to initial values. Vicitra
% change in comparison to Preliminary phase 13.46 ↓ 14.91 ↓ 14.46 ↓
Pratyayarabdha drug Bhumyamalaki shows 38.57% increase in water intake in comparison to initial values; however both the observations were found to be statistically non-significant. When the values were compared with control group, Nimba decreased water intake non-
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significantly and Bhumyamalaki increased water intake non-significantly (Table - 3). Fresh faecal output in all the three groups including control group was increased when the values of experimental phase were compared with preliminary phase. Nimba treated group shows 1.77% increase in fresh faecal output, while Bhumyamalaki administered group shows 46.24% increase in fresh faecal output. When the values were compared with control group, Nimba administered group marginally decreased faecal output while Bhumyamalaki group increased it apparently (Table - 4). In weight of dried faecal output all the groups shows increase when the values of experimental phase were compared with preliminary phase and the observed increase is almost similar to that of observation made in fresh faecal output (Table - 5). In control group 53.33% water content in faecal matter was observed. Nimba administered group showed only 25% water content, which is much lower than that of control group. In contrast, Bhumyamalaki treated group shows more faecal water content (Table - 6). Regarding number of faecal pellets, when the values were compared with control group, Nimba of Samana Pratyayarabdha group shows decrease in number of faecal pellet, while Bhumyamalaki of Vicitra Pratyayarabdha treated group shows increase in number of faecal pellets (Table - 7) In food conversion ratio, control group itself shows 13.46% decrease when the experimental phase value was compared with preliminary phase values. The similar type of decrease was also found in the Nimba and Bhumyamalaki administered groups (Table - 8). DISCUSSION: The present animal study was designed to assess the effect of drugs according to their Vipakas (post digestive effect) on Koshtha related parameters. According to Ayurvedic concepts, Madhura Vipaka (sweet post
digestive effect) causes Srishta Vinmutrata (easy excretion) i.e. increase in quantity and frequency of urine and stool, elevates Kapha dosha and increases Shukra dhatu, while Katu Vipaka (pungent post digestive effect) causes Baddha vinmutrata (difficulty in excretion) i.e. decrease in quantity and frequency of stool and urine, elevates Vata dosha and decreases Shukra dhatu (Acharya Yadavaji T , 2009d). Thus Bhumyamalaki, having Madhura Vipaka (sweet post digestive effect) should cause increase in stool and urine output and Nimba, having Katu Vipaka (pungent post digestive effect) should cause decrease in stool and urine output. As the stool and urine output are directly related to food consumption and water intake, parameters related to metabolic activity like weight change, food consumption, water intake and faecal output were also measured in the present study. Vipaka is not the only factor, which affects the metabolism, food consumption, faecal output etc. Ayurvedic pharmacodynamic properties like Rasa (taste), Guna (properties), Veerya (potency) and Prabhava (cause for specific action), also can affect these parameters. Both the drugs are having Tikta pradhana Rasa (bitter taste) and Sheeta Veerya (cold in potency). Nimba having laghu- snigdha guna (light and unctuous) and Bhumyamalaki having laghu- ruksha guna (light and dry property). Tikta rasa (bitter taste) have Deepana, Pachana, Lekhana, Mootra- Purisha Shoshana, Ruksha (dry), Laghu (light) and Sheeta (cold) properties. Sheeta Veerya (cold potency) has Stambhana property. In the present study dissimilarity between Samana and Vicitra Pratyayarabdha drugs was mainly at the level of Vipaka (post digestive effect). So it was supposed that drugs should show difference in action at the level of Vipaka (post digestive effect) only. Nimba is having Katu Vipaka (pungent post digestive effect), so supposed to decrease the faecal and urine output, food intake etc. Bhumyamalaki is having Madhura Vipaka (sweet post digestive effect), so supposed to cause increase in these parameters.
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Normal progressive increase in body weight was occurred in control group. In Nimba administered group, only 3 to 4 % increase in body weight was seen and the observed increase is comparatively less than that of control group. The observed change may be attributed to tikta rasa (bitter taste) of Nimba, which have Deepana â&#x20AC;&#x201C;pachana properties (Acharya Yadavaji T , 2009e). In contrast to this an apparent increase in body weight was observed in Bhumyamalaki administered group, the observed change may be attributed to Deepana and Rasayana properties of Bhumyamalaki (Mishra Shri Brahmashankara et al., 2010). Further, Bhumyamalaki is well established hepatoprotective drug and by virtue of these properties it may have enhanced activities of digestive enzymes. This may be also one of the reasons in observed weight gain in this group. Food consumption was increased in both treated groups, while in control group it was decreased, which shows that both drugs exhibit some of Deepana Karma. Food consumption is depends on Abhyavaharana Shakti (food intake capacity) and status of Agni (digestive power), so it can be said that all drugs helped in improve the status of Agni (digestive power), but the effect was not sufficient to reach up to significant level. Bhumyamalaki treated group shows better result because it has Ruksha Guna (dry property), which can improve Agni (digestive power) by decreasing Dravata (liquidity) of Pitta (Paradakara Hari S S, 2010b). The observed result in terms of water intake is according to Arabdhata as expected. Bhumyamalaki administered group showed increase in water intake when compared to other groups, it may be due to Ruksha Guna (dry property), as Rukshata (dryness) and Ushnata (hotness) are the two Gunas (property) which causes thirst due to Drava Shoshana property (Acharya Yadavaji T , 2009f), hence this may be the reason in observed increase of water intake. In contrast Nimba showed decrease in water intake the reason may be Snigdha Guna (unctuous property) of this drug.
The results of faecal output also substantiated the hypothesis of Arabtada. Dried faecal output depends on water content of stool, which evaporates during drying in oven. The dried faecal output was also found to be increased in all groups when data of experimental phase was compared to preliminary phase. In other words, the normal water content of faecal matter is about 53.33% as revealed in control group. In contrast, Nimba administered group showed only 25% water content, which is much lower than that of control group, while Bhumyamalaki treated group shows more faecal water content. The observed decrease in water content of Nimba administered group indicates that Nimba act according to its katu Vipaka (pungent post digestive effect) and shows baddha Vitkata (difficulty in excretion). Bhumyamalaki group, which showed increase in faecal water content, which is virtue of its Madhura Vipaka (sweet post digestive effect), it indicates that Bhumyamalaki causes Srishta Vitkata (easy excretion). These observations further corroborated by effect of test drugs on number of faecal pellets. Food conversion ratio is related to the Pachana property of drug. The percentage changes in all groups were very minute when compared to the initial values. It shows that drugs did not show any significant impact on Pachana. The reason involved may be short duration of drug administration adopted in this study. Though we expect hypothetically that Vipaka would cause the effect accordingly i.e. Srishta Vinmutrata (easy excretion) or Baddha Vinmutrata (difficulty in excretion), however always drug may not show desired effect as expected as the action of the drug depends on physiology and pathology of the individual and also on several factors like dose, potency of drug, Desha (place), Kala (time), proper digestion and metabolism etc. Acarya Nagarjuna states that, the Vipaka (post digestive effect) also changes according to the Dravya (substance), dose, Samskara (processing), Satmya (habits), Agni Bala
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(digestive power), Desha (place), Kala (time) and Samyoga (combination) (Narasimha, Sankara Menon, 1928). It is the peculiarity of Ayurvedic drugs that they act in pathological condition, but not show the same effect in normal physiological condition. For example, Sudarshana lowers the temperature in condition of fever, but if given in normal condition it doesn’t causes lowering of temperature. It is also very true with allopathic drugs also. Thus there is uncertainty in action of drug which depends on several conditions. Effect of Vipaka (post digestive effect), Veerya (potency) etc. will depend on the conversion of drug during digestion and metabolism at various levels. If at the level of Vipaka (post digestive effect) breakdown and re-synthesis doesn’t happen (Arabdhata remain unchanged), then drug will not show the effect at the level of Koshtha. It may act at the level of Dhatu (tissues) by the effect of Veerya (potency). If at the level of Veerya (potency) also, conversion will not take place then drug will act by the Prabhava (cause for specific action) or Rasa (taste) and Guna (property). Because action of Rasa (taste) and Guna (property) is sure and not depends on resynthesis or conversion during digestion.
Because of this uncertainty of Karma (action) at various levels it is said that, some substances act in accordance with their rasa (taste), others in accordance with their Vipaka (post digestive effect), and yet others in accordance with their Guna (property) or Veerya (potency) or Prabhava (cause for specific action) (Paradakara Hari S S, 2010c). CONCLUSION: In the chosen dose and duration of pharmacological study selected Samana and Vicitra Pratyayarabdha drugs showed apparent impact on Koshtha related parameters as per their Vipakas (post digestive effects) although the observed results are non-significant. Thus this study provides unequivocal basis to concept of Arabdata of Ayurveda. However the same set of drugs may be tried in higher doses as well as in longer duration to draw meaningful conclusion on related parameters and also cross study may be designed in which Madhura Vipaka (sweet post digestive effect) drugs would administered in condition of Baddhavitkata (difficulty in excretion) and Katu Vipaka (pungent post digestive effect) drugs would administered in condition of Srishtavitkata (easy excretion).
REFERENCES Acharya Yadavaji T (2009)a. Editor, Charaka Samhita of Agnivesha, Charaka & Dridhabala, Sutrasthana 26/58, with Ayurveda Deepika commentary of Chakrapani, Reprint ed, Chaukhamba Surbharati Prakashana, Varanasi. pp.146.
Acharya Yadavaji T (2009)c. Editor, Charaka Samhita of Agnivesha, Charaka & Dridhabala, Sutrasthana 26/61-62, with Ayurveda Deepika commentary of Chakrapani, Reprint ed, Chaukhamba Surbharati Prakashana, Varanasi. pp.146.
Acharya Yadavaji T (2009)b. Editor, Charaka Samhita of Agnivesha, Charaka & Dridhabala, Sutrasthana 26/48-52, with Ayurveda Deepika commentary of Chakrapani, Reprint ed, Chaukhamba Surbharati Prakashana, Varanasi. pp.146.
Acharya Yadavaji T (2009)d. Editor, Charaka Samhita of Agnivesha, Charaka & Dridhabala, Sutrasthana 26/61, with Ayurveda Deepika commentary of Chakrapani, Reprint ed, Chaukhamba Surbharati Prakashana, Varanasi. pp.146.
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Acharya Yadavaji T (2009)e. Editor, Charaka Samhita of Agnivesha, Charaka & Dridhabala, Sutrasthana 26/43, with Ayurveda Deepika commentary of Chakrapani, Reprint ed, Chaukhamba Surbharati Prakashana, Varanasi. pp.146. Acharya Yadavaji T (2009)f. Editor, Charaka Samhita of Agnivesha, Charaka & Dridhabala, Chikitsasthana 22/4, with Ayurveda Deepika commentary of Chakrapani, Reprint ed, Chaukhamba Surbharati Prakashana, Varanasi. pp.566. Bhattacharya Shri Taranatha Tarkavachaspati (2002). Vachaspatyam, part 1, Reprint Edition (2002), Chaukhamba Samskrita Series office, Varanasi, pp.795. Dhyani S.C.(2008). Rasa- Pancaka, chapter 3, Third Edition, Choukhamba Krishnadas Academy, Varanasi, pp.92. Dixit U.D, Dwivedi R.R. & Ravishankar B., (1995) ‘Fundamental concept of Panchamahabhuta and its utility in cikitsa’, Department of Basic principles, IPGT & RA, Jamnagar.
Narasimha, Sankara Menon (1928). Editor, Rasa Vaisheshika Sutra of Bhadanta Nagarjuna, 4/55, Superintendent, Government Press, Trivandrum. pp.197. Paradakara Hari S S (2010)a. Editor, Ashtanga Hridaya of Vagbhata, Sutrasthana 9/27, with commentaries of Sarvangasundara of Arunadatta & Ayurvedarasayana of Hemadri, Reprint Edition, Chaukhamba Sanskrit Sansthana, Varanasi, pp.172. Paradakara Hari S S (2010)b. Editor, Ashtanga Hridaya of Vagbhata, Sutrasthana 12/11, with commentaries of Sarvangasundara of Arunadatta & Ayurvedarasayana of Hemadri, Reprint Edition, Chaukhamba Sanskrit Sansthana, Varanasi, pp.193. Paradakara Hari S S (2010)c. Editor, Ashtanga Hridaya of Vagbhata, Sutrasthana 9/2223, with commentaries of Sarvangasundara of Arunadatta & Ayurvedarasayana of Hemadri, Reprint Edition, Chaukhamba Sanskrit Sansthana, Varanasi, pp.170.
Laurence, D.R. and Bacharach, A.L (1964). Editors, Evaluation of drug activities, In: Pharmacometrics by Paget, G.E., Barnes, J.M.., Vol. 1, Academic press, New York. pp.161.
Paradakara Hari S S (2010)a. Editor, Ayurvedarasayana of Hemadri, Ashtanga Hridaya Sutrasthana 9/27, with commentaries of Sarvangasundara of Arunadatta & Ayurvedarasayana of Hemadri, Reprint Edition, Chaukhamba Sanskrit Sansthana, Varanasi, pp.172.
Mishra Shri Brahmashankara and Vaishya Shri Rupalalaji, (2010). Editors, Bhavaprakasha Nighantu of Bhavamishra, Guducyadi Varga/ 278, including Bhavaprakasha Nighantu portion and Vidyotini Hindi Commentary, part 1, 11th ed. Chaukhamba Sanskrit Bhawan, Varanasi. pp.460.
Paradakara Hari S S (2010)b. Editor, Ayurvedarasayana of Hemadri, Ashtanga Hridaya Sutrasthana 9/27, with commentaries of Sarvangasundara of Arunadatta & Ayurvedarasayana of Hemadri, Reprint Edition, Chaukhamba Sanskrit Sansthana, Varanasi, pp.172.
Source of Support: Nil
Conflict of Interest: None Declared
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 629–638 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Review article PARKINSONISM IN AYURVEDIC PERSPECTIVE, A BIRD’S EYE VIEW Nayak Annada Prasad1* 1
Assistant professor, Kayachikitsa, MSM Institute of Ayurveda, B.P.S Women University, Khanpur kalan, Sonipat, Haryana, India *Corresponding Author: E-mail: vd_apnayak@rediffmail.com; Mobile: +919355506222
Received: 31/08/2012; Revised: 5/10/2012; Accepted: 24/10/2012
ABSTRACT Since ancient times, humans suffered from many neurodegenerative disorders, where he/she has to live with it. Parkinsonism is one of such neurodegenerative disorders for which no clear description is found in Ayurvedic texts. However the diseases, vepathu and Kampa Vata simulate with the Parkinson’s disease. According to some Ayurvedic experts Kampa (tremor), Sthambha (rigidity), Chesta sanga (bradykinesia), Vakgraha (dysphasia), and Smriti ksaya (dementia) are the cardinal features of kampaVata / vepathu. As per Ayurvedic description these symptoms are due to Vata prakopa (vitiation of Vata). This is an apatarpanotha (due to malnutrition) and pakwasayotha vyadhi (disease caused due to vitiation of dosas in the large intestine) where Dosas (three basic constituents of the body such as Vata, Pitta, Kapha) are located in Gambhira sthana (in deep seated organs). So it needs strenuous management strategies unless it becomes incurable by time. Snehana (oleation), Svedana (sudation), Anuvasana Basti (oily enema), Niruha Basti (decoction enema), Shiro Basti (Procedure to retain oil on scalp) and Virechana (purgation) may be planned in the management of Kampa Vata. KEY WORDS: Parkinsonism, Kampa, Sthambha, Vata prakopa, Vepathu.
To Cite this article: Nayak Annada Prasad (2012), PARKINSONISM IN AYURVEDIC PERSPECTIVE, A BIRD’S EYE VIEW, Global J Res. Med. Plants & Indigen. Med., Volume 1(11), 629–638
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INTRODUCTION Parkinsonâ&#x20AC;&#x2122;s disease is a chronic, progressive degenerative disorder of the central nervous system usually seen in 6th decade of life. It is characterized by the symptoms such as tremor, rigidity, akinesia / bradykinesia and postural instability1. These symptoms are also associated with gait and speech disturbances, difficulty in swallowing; sleep disturbances, depression and dementia in course of the disease2. These symptoms may appear slowly and in no particular order. At early stage, symptoms may be subtle and progress over many years before reaching a point where they interfere with normal daily activities2. In Charaka Samhita (an ancient book written by Charak) it is described that diseases are innumerable in number depending on their distinctive features like signs and symptoms, etiology, site of origin, manifestation etc. One Dosa (three basic constituents of the body such as Vata, pitta, kapha) makes different diseases in relation with the difference in etiological factors (sthanantaragatava). Therapeutic initiation can be achieved by understanding the nature of disease (vikaraprakriti), various sites of morbid dosa (three basic constituents of body), different locations (adhisthanantarani) and variety of causes (samuthananvisesa)3. The exact description of Parkinsonâ&#x20AC;&#x2122;s disease, as a single disease, is not available in Ayurvedic classics. Certain symptoms are found under Sirokampa4, Vepathu5. In Charaka samhita, Sirokampa (head tremor) is enlisted as one among the important Sirorogas (diseases of head) like Ardita or 3 Ardhavabhedak (facial palsy). Sirokampa caused by vitiation of Vata, due to aggravation of Ruksha guna (ununctuous property) is explained6. Vepathu is enlisted fewer than one among the 80 Nanatmaja vikaras of Vata7 (exclusive Vataja disorders). In the text of Madhava Nidana, Vepathu has been defined as generalized Vata vikara (diseases of Vata) with sarvanga kampa (generalized tremor) 8 including sirokampa (head tremor). In the text Basavarajiya, a separate disease with the nomenclature as kampaVata (tremor due to
vitiation of Vata) has been referred and different varieties of kampa (tremor) have been explained as kakaVata (disease of Vata as in crow) and bahukampaVata9 (extreme tremor). Development of signs and symptoms as per Ayurvedic pathophysiology Ayurveda has its own way of describing patho-physiological conditions of different diseases. Vata (one of the three basic constituents of the body) is the one which controls all the motor activities (cesta)10. Among the five divisions of Vata (prana, udana, samana,vyana, apnana), vyanavayu is responsible for body movements (chesta) and gait (gati)11. Cesta (motor activities) as well as Gati (movement) is contributed by moving (chala) property of Vata. Charaka has omitted chala (movement) from the properties of Vata and instead, included daruna10 (severe). Here, Acharya Cakrapani, observed darunatva as chalatva (movable) itself because of chalatva (movable property). He also puts forward a second opinion that darunatva (severity) is reducing in nature (sosana svabhava). The second classification seems to be more appreciable. The chala (moving) is more or less an action (karma) rather than a property (guna) especially in therapeutic aspect. The chala property can be manipulated by applying other properties like cold (Sita), hot (Usna), light (Laghu) and heavy (Guru). In therapeutic aspect hypo actives (chal abhava) or hyper actives (chaladhikya) can be treated by Snigdhosna (oily and hot preparations) or Rukshasita prayogas (Ununctuous and cold preparations). Vataprakriti (person with nature of Vata) individuals are intolerable to cold and show frequent tremors and rigidity13. Interdependent kampa and stamba are the abnormal manifestations of chala. Kampa is manifested when sitaguna (cold property) gets in association with rukshadi gunas (ununctuous properties). When sitaguna (cold property) combines with snigdha (oily property), stambha 12 (rigidity) may manifest.
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Here certain considerations of upadhatu (body constituents like ligaments and tendons) are also important. Snayu (ligaments) is explained as upadhatu of meda14 (fatty tissues). Ruksana (dryness) or Snehana (oilyness) directly operates on medodhatu (fatty content in body) and subsequently shows influence on its upadhatu (snayu). Medososana (absorption of fatty content) or medovrddhi (raised level of
fatty content) may manifest as snayuvikara (diseases of ligament). Analysis of Symptomatology of Parkinson’s disease in Ayurvedic Perspective To identify the nature of disease, the important symptomatology of Parkinson’s disease is detailed below with its possible Ayuredic analogues (Table 1):
TABLE -01 COMPARISON OF SYMPTOMATOLOGY Sr. No. 1 2 3 4 5 6 7 8 9 10 11
Symptomatology of Parkinson’s disease2
Ayuredic analogues
Tremor Rigidity Bradykinesia Akinesia Gait disturbance Postural instability leading to fall Dysphasia Dysarthria Flexed posture Dementia Depression
Kampa Stambha Chestasanga Chestahani Gatisanga Skhalanam gatau Vakgraha Svaragraha Vinamana Smrtiksaya Visada
The Ayurvedic terminologies used above are not simple translations, but are taken from different contexts of Ayurvedic classics, amongst the above symptoms, Kampa and stambha (rigidity) are important. They are preliminary and the others are resultant of the same. Hence, these two symptoms have to be analyzed in detail.
of pitta and increased level of pitta and kapha). According to Astanga-samgraham (an ancient book written by Vagbhata), when pitta is ksina (less), asayapakarsa (movement of dosa from its original site) of samakapha (normal kapha) with kupitta Vata (aggravated Vata) cause stamhba along with Saitya (coldness) and Gaurava (heaviness).
Stambha (rigidity)
Stamhba and kampa occurs when Vata is vitiated in snayu15 (ligament). When dosas are located in snayu, sira (arteries) or kandara (tendons), the symptomatology is stambha, sankocha (contraction), khalvi (cramps), granthi sphurana (pining sensation) or supti19 (numbness). Stamba may manifest as localized (ekanga) or generalized (sarvanga). It is a symptom in mamsamedogata-Vata16 (vitiated Vata in fleshy and fatty tisses), snayupra Vata15 (Vata in ligament). sarvanga Vata16 (Vata located in whole body) and sirastha kapha (kapha located in head). The above points are summarized in table 2.
Stambha is defined as stabdhata16 or nischalikaran15 (suppression of movements). It can be explained as rigidity or stiffness. Stambha is a cardinal symptom17, which is also seen in pittaksaya17 (decreased level of pitta). While enlisting pathology of different Dosavikalpa (combination of dosas), Charaka explained stambha and vepathu along with other symptoms like Saitya (feeling of coldness), Toda (pricking type pain), Gaurava (heaviness), etc. in the symptomatology of hinapitta-vrddha Vatakapha18 (decreased level
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TABLE -02 SAMPRAPTI GHATAKA OF STAMBHA Sr. No. 1
2 3 4 5
Samprapti ghataka
Samprapti
Dosa (Vata, Pitta, kapha)
Vatakopa (Vata ↑↑)
Dhatu (tissues like blood, flesh etc.) Upadhatu (arteries, ligaments, and tendons) Dehadosa (Vata, pitta, and kapha in body) Specific Pathology
Pittaksaya (pitta ) Hinapitta- kaphaVatavridhavikalpa (pitta Vata ↑ kapha ↑ ) Ksinapitta asayapakarsana of Samakapha by vriddha Vata Pitta kapha = Vata ↑) Mamsa, meda (fleshy and fatty tissues) Snayu (ligaments) Ekanga/Sarvanga (localized or generalized) Mamsamedogata Vata (Vata situated in fleshy and fatty tissues) Snayuprapta Vata (Vata in ligaments) Sarvanga Vata (generalized Vata) Sirastha kapha (kapha located in head)
Kampa (tremor) Kampa is explained as Vepathu or Ativepanam (excessive tremors). Localized obvious tremor is called vepanam20. Kampa may be correlated to tremor. It is a cardinal symptom of Vatavrddhi (raised level of Vata) as well as Kopa19 (aggravation). Vepana is explained under the symptom of kaphaksaya (decreased level of kapha). As explained earlier, vepana is seen along with stambha in dosavikalpa of hinapitta-vriddhaVatakapha17 (decreased pitta and increased Vata and kapha). Vepana may also manifest when pittaslesmaksaya (decreased level of pitta and kapha) leads to kevala Vatavrddhi (only raised Vata) which afflicts mamsa (fleshy organs). Rasaksaya (loss of lymph or bodily fluid) may be manifested as kampa17. Kampa is a cardinal symptom of snayuprapta Vata15 (Vata in ligaments). As in case of stambha, kampa may also be manifested as ekanga (localized) or sarvanga (generalized). The textual details
of kampa can be summarized as follows (table 3). Kampa and Stambha (tremor and rigidity) Kampa and stambha are abnormal patterns of chala (movement), and both seem to be interdependent. Kampa and stambha manifest together in hinapitta with kaphaVatavriddhi (decrease pitta and increase kapha-Vata). Vata is always vriddha (increase) or kupitta (aggravation) in both kampa and stambha. Pitta is almost always ksina (less). The status of kapha may be vriddha (increase), sama (normal level) or ksina (less). When Vata is vitiated in snayu, it may be manifested by diminishing the movements as in stambha or by hyperkinetic movements as in kampa or akhsepa (convulsion). Charaka has enumerated stambha and kampa as upadhatupradosajavyadhis21 (the sub-division of dhatus i.e. updhatus generated diseases).
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Cestasanga movements)
and
Gatisanga
(loss
of
Chestasanga, chestahani or kriyasu asakti (inability to do work) can be explained as bradykinesia or akinesia. Kriyasu asakti is defined as the inability to perform general body movements like aksepana (flexion), apaksepana (extension), prasarana (expansion), akunchana (contraction), etc. Chestastambha (loss of movement) is a symptom of kaphavrita vyana22 (Vata covered by kapha). Chestahani (loss of movement) as a symptom udanavrita vyana23 (vyana Vata
covered by udana). When vyana vata (type of vata) is subjected to avarana (covered by other), it fails to perform its functions like flexion, extension, etc. and there by slows down bodily activities. Diminished movements may be in the form of chestasanga (bradykinesia) or in the form chestahani (akinesia). Gati sanga can be explained as gait disturbances. It is a symptom of kaphavrita vyana24 (vyana Vata covered by kapha). Yanasakti (inability to move or travel) is seen in snayu marmavidha25 (injury on ligament and vital organs).
TABLE -03 SAMPRAPTI GHATAKA OF KAMPA Sr. No. 1
Samprapti (constituents pathogenesis) Dosa
2 3 4 5
Dhatu Upadhatu Dehadesa Specific Pathology
ghataka of
Samprapti (pathogenesis)
Vatavriddhi or Prakopa (increase or vitiation of Vata) Pittaksaya (pitta ) Kaphaksaya (kapha) Hinapitta-vriddha Vata (decrease pitta and increase Vata), Kaphavikalpa (combination with other) Ksinapitta-kaphaVata- vriddhavikalpa (decrease pitta combination with increase kapha-Vata) Rasa ksaya (decrease in lymph or bodily fluid) Snayu (ligaments) Ekanga/Sarvanga (localized or generalized) Snayuprapta Vata (Vata in ligaments) Sarvanga Vata ( generalized Vata)
Involvement of higher functions Vakgraha may be compared with dysphasia. Svaragraha is dysarthria. Vakgraha is an important symptom of kaphavrta vyana (Vata covered by kapha). Vakgraha (dysphagia) as well as svaragraha (dysarthria) are together seen in kaphavrta udana26 (udana Vata covered by kapha). In the advanced stage of this disease, prana Vata (type of vata) may be associated in pathology and manifested as symptoms with disturbances in memory, mood and cognition as smrtiksaya (dementia) or visada (depression). Smrtiksaya (dementia) and sarvendriyanam sunyata (sensorial
impairment) are symptoms of pranavrtavyana (vyana Vata covered by prana). Samuthanavisesa/ Causative factors Comprehensive knowledge on etiological factors (nidana parivarjana) is necessary for prevention of any disease (svasthyasamraksana) to get an idea regarding extent of dosadusti (vitiation of dosa) and to plan the line of management (upasayanupasaya). The different etiological factors explained in the context of Parkinsonism are listed below: 1. Idiopathic 2. Toxin induced
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3.
4. 5. 6. 7.
(Manganese, co-poisoning drugs) Drug induced (E.g. prolonged use of tranquilizing drugs) Repeated head trauma Accompanying other neurological disease Post encephalitic Tumors of brain (rarely)
Let us now examine the different etiological factors explained in Ayurvedic scriptures in clinical conditions similar to Parkinsonism. Administration of Rukshadi (Ununctuous properties)
Guna
Charaka highlights sirokampa (tremor of head) in the context of Trimarmiya siddhi (a chapter in Charaka’s work on Ayurveda) and explained it, as caused by kupitta Vata (vitiation of Vata) due to administration of rukshadi gunas. It may be understood as aggravation of gunas like ruksha (unctous), laghu (lightness), khara (roughness) etc. along with sitaguna (coldness). Substances containing kasayarasa (astringent in taste) impart stambha (stiffness). Tea, coffee, narcotics, chewing of tobacco, etc. may be identified as kasayarasa-pradhanadravyas (plants with astringent taste). Katurasa (pungent in taste) imparts kampa (tremors) in the body. Pickles and spicy foods contain too much katurasa. Rukshadi dietary articles may be understood as fried food, pulses like kalaya, rajamasa, etc. tubers, wafer, biscuits, fast food and soft drinks. Chinta (tension), shoka (sadness) and bhaya (fear) leading to continuous stress, are important viharas (daily activities) for aggravation of rukshadiguna by rasaksaya (loss of lymph or bodily fluids). Smoking also aggravates rukshadiguna. The same way air conditioning or continuous exposure to cold air (sitamarutaseva) provokes Vata. Visa Sushruta said that Stambha and kampa are important symptoms of Dusivisa (animal
poison) as well as Sthavaravisa (plant poisons). According to Sushruta, kalakuta (Aconite) visa causes vepathu (tremor), stambha (stiffness) and sparsanjana (anesthetic effect)27. Musthaka (Cyperus rotondus) is also a deadly poison enlisted under sthavaravisa which causes gatrastambha (stiffness in body) and vepathu27. Sirobhighata and svabhavaja Caraka has not explained stambha (Stiffness), kampa (tremors) or any other related symptoms among symptomatology of sirobhighata (injury in head region), but the context is explained as acute head trauma. Parkinson’s disease is not the resultant of acute head trauma, however repeated head trauma certainly causes the same, like Punch Drunk Syndrome as in the case of professional boxers. In this context, a dosaja marmabhighata (injury caused in the vital parts by dosha) of siromarma (vital part head), which is common in target aiming professionals like businessmen, etc. who has overload of files, may be considered. Iatrogenic causes Over administration of stambhana therapy (sluggishness inducing property) leads to stamba, sankocha (contraction) of snayu and tvak (skin), kampa (tremors), hritgraha (tightness of chest), vakgraha (dysphasia) and hanugraha28 (lock jaw). This can be included as an iatrogenic cause of the disease. Samprapti (etipathogenesis) The etiological factors, symptomatology, site of manifestation of Parkinsonism have been discussed earlier. From these observations, the samprapti (pathogenesis) of Parkinson’s disease has been formulated on Ayurvedic parlance. The clinical condition is essentially caused by Vatakopa (vitiation of Vata). Vata may get vitiated by dhatuksaya (loss of body tissues) or avarana (one dosa covered by the other). In the first case, administration of Rukshadi gunas (ruksha, laghu, khara, etc.) along with
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sitaguna (coldness) causes dhatuksaya and following riktata (emptiness) in dhatu (body tissues) and correspondingly in their respective srotas (channels). In the case of avarana, the etiopathological factor is that it aggravates the kapha. Kapha is aggravated along with Vata along with qualities like usna (hot), snigdha (oilyness), sushka dravyas (dry materials) and guru gunas (heavy property) combined with sita (cold) just as in urustambha (rigidity of thigh). The aggravated kapha cause obstruction (margarodha) of Vata, very specifically vyanaVata. In the pathological sequence of dhatuksaya, Vata gets more space (avakasa) in raktadhatu (blood) or srotas and vitiated Vata with increased chala property is called as gataVata. The vitiated Vata, because of specificity of nidana (etiology) as well as khavaigunya (vitiation of dosa in empty space), identifies its adobe as snayu and manifests as snayuprapta Vata (vitiation of Vata in ligaments). Snayuprapta Vata is characterized as stambha and kampa. They seem to be intercomplimentary. Stambha contributes kampa and vice versa because of the same adhisthana (seat) of vitiated dosas. In the pathological sequence of avarana of Vata by kapha, it may manifest generally as kaphavrita Vata (Vata covered by kapha) and shows symptoms of localized or generalized stambha. The avarana of Vata by kapha may be very specific to kaphavrtavyana (vyana covered by kapha). The kaphavrta vyana shows the symptoms of chestasanga, gatisanga, skhslanam gatau, etc. which are nothing other than due to poverty of movements. The pathology of avarana of kapha may advance to higher functional levels of Vata like kaphavrtaudana (udana Vata covered by kapha), which manifest as vakgraha (dysphasia) or svaragraha (dysarthria). These are speech disturbances characterized as dysarthria and dysphasia. When udana and vyana are under the avarana of kapha, it may further cause anyonyavarana (type of avrana) of udana and vyana. Uddanavrtavyana (Udana
covered by vyana) is manifested as chestahani (akinesia). The anyonyavarna (cover by each other) may further advance to higher functional levels according to worsening of pathology. When vyana get avarana by prana (i.e. pranavrtavyana), loss of memory (smrtiksaya), loss of motor power (balaksaya) and impaired sensorium (indriyanam sunyata), may manifest. Probable management scheme of kampaVata/vepathu After analysing the pathophsiology of kampaVata, Snehana (oleation), Svedana (sudation), Anuvasana Basti (oily enema), Niruha Basti (decoction enema), Shiro Basti (procedure to retain oil on scalp) and Virechana (purgation) may be planned in the management of KampaVata. Some of the important recipes for KampaVata are Triphaladi Mahasneha, Sahachara Taila, Chaturbhuja Rasa, Maharasnadi Yoga may be prescribed. Experimental studies have proven the effect of some Ayurvedic drugs like Kapikacchu29 to show significant antiparkinsonism effect. Basti (enema) extirpates vitiated Vata. It is thus the chief therapy for the diseases of Vata. As described earlier in the Samprapti (pathogenesis), KampaVata may be considered as one of the disease of provoked Vata due to Dhatukshaya (loss of tissues) and Avarana (cover). Yapana Basti30 (type of enema) is also indicated in Avarana (cover) and Basti (enema therapy)is indicated for Vepathu (tremors) by Acharya Charaka. As mentioned earlier KampaVata is a disease of elderly people which is dominated by Vata. Yapana Basti (type of enema therapy) being Balya (nourishing), Rasayana (rejuvenating), Mrudu (mild) in nature and it suits the above mentioned conditions. CONCLUSION Vepathu and Kampavata are apatarpanotha vyadhi (disease of malnutrition) and pakvasayotha vyadhi (disease caused by the vitiation of dosas in large intestine). It is an upadhatupradosaja vikara (disease produced in the subdivision of dhatus) and marmastha
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vyadhi (disease of vital organs). Due to the seriousness of the location of dosa (gambhirya of sthana), even in the earlier stage (nava), the disease is krcchratama asadhya (prognosis only on strenuous management strategies), otherwise the disease becomes incurable by time. To obtain satisfactory results, the management should be in such a way that it should bridle the aggravated Vata, Dhatukshaya (loss of tissues) and Avarana (cover). This goal can be achieved only by the use of Rasayana drugs (rejuvenating medicines) in Basti (enema) form. Moreover, Basti (enema) also promotes the longevity and provides stability to Dhatus. Mustadi Rajayapana31 (type of decoction enema) has been said to be Shukrajanana (spermatogenic effect), Mamsajanana (increase body built), Balajanana (increases energy) by Maharshi
Charaka. Moreover all the indications of this Basti suggest that it may be useful in Avarana (cover) as well as in Dhatukshaya (loss of dhatus) which are the two main events in the pathogenesis of KampaVata. Mustadi Rajayapana (type of decoction enema) is the best mode of administration of Rasayana drugs and possesses all the qualities of Basti mentioned earlier. After analyzing the symptomatology of Parkinsonism, we can say that this can be compared with vepathu, sirokampa and kampavata. Whatever the comparison, the treatment should be planned after governing the pathophysiology of parkinsonism which must be compared with dosage and dhatu kshaya vriddhi lakshanas (symptoms of vitiated dhatus) and avarana vata lakshana (symptoms of covered vata).
REFERENCES 1. Edwards, Bouchier, Haslet, Chilvers, (1995) seventh edition, Davidson’s principle and practice of medicine, ELBS, chapter -18, p.- 1079 2. Harrison, (2008), 17th edition, Harrison’s principle of internal medicine, McGrawHILL, Volume-II, , chapter-366, p.- 2549 3. Agnivesh, Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Sutra sthana -18/45, Chaukhamba Surabharati Prakasana, Varanasi, p.- 378 4. Agnivesh, Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Sutra sthan-17/14, Chaukhamba Surabharati Prakasana, Varanasi, p.- 335, 5. Agnivesh, Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Sutra sthan-20/11, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 390,
6. Agnivesh, Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Sutra sidhisthana -9/86, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 1293, 7. Agnivesh, Charak, (1995), 4th edition, Charak Samhita Charak, Chandrika Hindi Byaksha, Sutra sthana -20/11, Edited by Brahmananda Tripathy, Chaukhamba, p.690 8. Madhava kara, (1993), 22nd EdiMadhava Nidan, part-1,22/74, Madhukosha Sanskrit commentary by Sudarsana Sastri, Chaukhamba Sanskrit Bhavan, p.- 447 9. Basavraj, (1987), Basvarajiyam, Ed and Pub. By Rajeshwar Datt, Shastry, Chalikhambha sanskrit series, varanasi. 10. Agnivesh, Charak, (1995), 4th edition, Charak Samhita Charak, Chandrika Hindi Byaksha, Sutra sthana -12/8 and 12/4, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 253
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11. Bridha Vagbhataa, (1996), Astanga sangraha, Sutra sthan-20/4, Soroj Hindi commentary, Edi. By Ravidutta Tripathy , Chaukhamba Sanskrit pratisthana, Delhi, p.- 375
19. Agnivesh Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Sutra sthan-28/22, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 549
12. Vagbhata, (1997), Astanga Hridaya, Vidyotini bhasa Teeka by Kaviraj Atridev Gupt, Sutra Sthana, 9/19, twelfth edition, Chaukhamba Sanskrit bhawan, Varanasi, p.- 81
20. Agnivesh Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Chikitsa sthan-24/102, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 820
13. Agnivesh, Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Viman sthana -8/98, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 761.
21. Sushrutha, (1997), 11th edition, Sushrutha Samhita, , Ayurveda Tatwa Sandeepika Hindi Commentary, Edited By Kaviraj Ambikadatta Shastree, Nidana sthana.1/39, Chaukhamba Sanskrit bhawan, Varanasi, p.- 231.
14. Agnivesh, Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Chikitsa sthana 15/17, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 553
22. Agnivesh Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Chikitsa sthana -28/214, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 976
15. Sushrutha, (1997), 11th edition, Sushrutha Samhita, , Ayurveda Tatwa Sandeepika Hindi Commentary, Edited By Kaviraj Ambikadatta Shastree, Nidana sthana.1/27, Chaukhamba Sanskrit bhawan, Varanasi, p.- 230
23. Agnivesh, Charak, (1995), 4th edition, Charak Samhita Charak Chandrika Hindi Byaksha, Chikitsa sthana -25/228, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 979.
16. Vagbhata, (1997), twelfth edition, Astanga Hridaya, Vidyotini bhasa Teeka by Kaviraj Atridev Gupt, Nidana Sthana, 15/11 Chaukhamba Sanskrit bhawan, Varanasi 1997, p.- 276, 15/15, 277
24. Vagbhata, (1997), 12th edition, Astanga Hridaya, Vidyotini bhasa Teeka by Kaviraj Atridev Gupt, Sarira Sthana, 4/48, Chaukhamba Sanskrit bhawan, Varanasi, p.- 196
17. Bridha Vagbhataa, (1996), Astanga sangraha, Sutra sthan-19/9 and 19/10, Soroj Hindi commentary, Edi. By Ravidutta Tripathy , Chaukhamba Sanskrit pratisthana, Delhi, p.- 362
25. Agnivesh Charak, (1995), 4th edition, Charak Samhita, Charak Chandrika Hindi Byaksha, Chikitsa sthana -28/224, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 978
18. Agnivesh, (1995), 4th edition, Charak, Charak Samhita ,Charak Chandrika Hindi Byaksha, Sutra sthan-17/56, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 348,
26. Sushrutha, (1997), 11th edition, Sushrutha Samhita, , Ayurveda Tatwa Sandeepika Hindi Commentary, Edited By Kaviraj Ambikadatta Shastree, Kalpa sthana.2/150,, Chaukhamba Sanskrit bhawan, Varanasi, p.- 20.
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27. Vagbhata, (1997), 12th edition, Astanga Hridaya, Vidyotini bhasa Teeka by Kaviraj Atridev Gupt, Sarira Sthana, 17/20, Chaukhamba Sanskrit bhawan, Varanasi, p.- 114. 28. Vagbhata, (1997), 12th edition, Astanga Hridaya, Vidyotini bhasa Teeka by Kaviraj Atridev Gupt, Chikitsa Sthana, 22/51-52, Chaukhamba Sanskrit bhawan, Varanasi, p.- 426 29. Dhurve sanjay (2001) : a clinical study on kampaVata and its management with
Source of Support: Nil
kapikachchhu, g.a.u., dept. of kayachikitsa, Jamnagar. 30. Agnivesh, (1995), 4th edition, Charak, Charak Samhita Charak Chandrika Hindi Byaksha, Chikitsa sthana -28/240, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 981. 31. Agnivesh, Charak, (1995), 4th edition, Charak Samhita Charak Chandrika Hindi Byaksha, Chikitsa sthana -28/240, Edited by Brahmananda Tripathy, Chaukhamba Surabharati Prakasana, Varanasi, p.- 981.
Conflict of Interest: None Declared
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 639â&#x20AC;&#x201C;643 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Research article SCOPE OF THIN LAYER CHROMATOGRAPHY IN QUALITATIVE EVALUATION OF AYURVEDA DRUGS Rajendra H M1*, Lad Meenal D 2 1
Assistant Professor, Sri Jayendra Saraswathi Ayurveda College, Nazarathpet, Chennai, Tamil Nadu, India. Professor & Ph.D.Guide, College of Ayurveda & Research Centre, Akurdi, Pune, Maharashtra, India. *Corresponding Author: E-mail: ayurrajhm@gmail.com; Mobile: +919952063158 2
Received: 14/09/2012; Revised: 24/10/2012; Accepted: 30/10/2012
ABSTRACT The World Health Assembly in resolutions has emphasized the need to ensure the quality of medicinal products by using modern control techniques and applying suitable standards. Thin Layer Chromatography comes with priority in ensuring proper quality of plant materials; by its separation technique is very useful in quality evaluation of single drug and compound drug preparations. In the present study single drugs like Shunthi, Maricha and Pippali, compounded drugs like Trikatu, Hingwashtaka churna and Sutashekara rasa were taken for quality evaluation through Thin Layer Chromatography technique. Genuine raw drugs of Shunthi, Maricha, Pippali, Trikatu were procured and Thin Layer Chromatography was performed to obtain Rf values which were considered as standards which denotes the quality and purity of the drug. Market test samples of Hingwashtaka churna and Sutashekara rasa were procured and Thin Layer Chromatography was performed to separate the compound mixture and to identify the presence of Trikatu in them by comparing the Rf values of Trikatu with Rf values of test samples. Results of the study conclude that Thin layer Chromatography is useful in Identification, Standardization and Qualitative evaluation of Ayurveda drugs. KEY WORDS: Thin Layer Chromatography, Rf Value, Shunthi, Maricha, Pippali.
To Cite this article: Rajendra H M, Lad Meenal D (2012), SCOPE OF THIN LAYER CHROMATOGRAPHY IN QUALITATIVE EVALUATION OF AYURVEDA DRUGS, Global J Res. Med. Plants & Indigen. Med., Volume 1(11), 639â&#x20AC;&#x201C;643
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INTRODUCTION Since ancient times, Vaidyas (Physicians) had identified the medicinal plants, collected, processed, formulated and applied themselves. These plant drugs have stood the test of time for their safety, efficacy, cultural acceptability and lesser side effects. About 75–80% of the world population, mainly the developing countries still are using the plant-based medicines for primary health care. Quality of the plant-derived medicines is a matter of great concern as the utilization of plant materials to cure infections and chronic diseases are increasing. Hence it becomes necessary to assure the consumer and prescriber the authenticity of raw material for its consistency, quality and efficacy. (Kokate C.K. et al., 2005). The solution for this is the standardization techniques and methods, which are useful for the quality evaluation of medicinal plant materials. Even the “World Health Assembly” in resolutions has emphasized the need to ensure the quality of medicinal plant products by using modern control techniques and applying suitable standards. Thin Layer Chromatography is one of the most authentic, cost effective and easy to perform method with less expensive equipment and the technique is frequently used for evaluating medicinal plant materials and their preparation (Anonymous, WHO, 2002). MATERIALS AND METHODS Thin Layer Chromatography (TLC) is a separation technique used for the separation of compounds of mixture by their continuous distribution between two phases, one of which is moving past the other. It works on the basis of Adsorptive principle. The Retention factor (Rf) value of TLC is specific to solute and solvent system of that specific drug. This is characteristic chromatography pattern of every individual plant (Chatwal and Anand, 1996). Sample selection – To study the scope of TLC with respect to
Ayurveda drugs, single drugs and compound drugs were selected. Single pungent drugs like - Shunthi (Rhizome of Zingiber officinale Rosc), Maricha (Fruit of Piper nigrum Linn) and Pippali (Fruit of Piper longum Linn). Compound drugs like – Trikatu (a compound containing 3 pungent drugs), Hingwashtaka churna (A Medicinal formulation in the form of powder) and Sutashekara rasa (A Medicinal formulation in the form of pills) (Chunekar and Pandey, 2006). Raw drug samples of Shunthi, Maricha and Pippali were authenticated by Agharkar Research Institute; Pune and Voucher specimens were deposited at the Herbarium of Sri Jayendra Saraswathi Ayurveda College, Chennai. The drugs were made into coarse powder. Equal quantity of the individual powder was taken separately and mixed thoroughly to prepare a compound drug Trikatu. Other compound drugs like Hingwashtaka churna (Formulation containing Zingiber officinale Rosc, Piper nigrum Linn, Piper longum Linn, Apium graveolens Linn, Sodii chloridum, Cuminum cyminum Linn, Carum carvi Linn, Ferula foetida Regel) and Sutashekara rasa (Formulation containing Zingiber officinale Rosc, Piper nigrum Linn, Piper longum Linn, Purified Mercury, Borax, Purified Aconitum ferox Wall, Datura metel Linn, Purified and processed Sulphur, Bhasma prepared from Copper, Bhasma of Conch Shell, Elettaria cardamomum Maton, Cinnamomum zeylanicum Blume, Cinnamomum tamala Nees & Eberm, Mesua ferrea Linn, Aegle marmelos Corr, Curcuma zedoaria Rosc, juice extract of Eclipta alba Hassk) of Dabur and Baidyanath companies were collected from the drug house (Mishra Shri Brahmashankar, 2006). Procedure – Extracts of the above drugs were prepared separately by taking 1 gm of powder and 10 ml of alcohol by cold maceration. Fine slurry of silica gel with binder Calcium Carbonate (stationary phase) was prepared in a beaker with distilled water. The slurry was then prepared on standard glass plates with the help of spreader and a thin layer was prepared about 250 µm thickness. These plates were made to
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dry in air for 30 min, then in an oven at 110°C for another 30 min and allowed to cool, which makes the adsorbent layer active. By using micro capillaries the spots of the extract of above drugs not more than 4 mm in diameter were placed on to the starting line of the plates, about 15 mm above the lower edge. Then the spots were allowed to dry. In the proportion of 93:7, Toluene and Ethyl acetate (Mobile phase) was taken. 10 ml of it was poured into separate chromatographic chambers. Then the chambers were closed and allowed to stand for at least 1 h at room temperature to achieve saturation of the chamber, in order to avoid unequal solvent evaporation losses from the developing plate. Then the plates were placed into the chromatographic chambers ensuring that the spots were above the surface of the mobile phase. Then the chambers were closed and the solvents were allowed to flow by ascending type. The chromatograms were allowed to develop at room temperature (Chatwal and Anand 1996). After the mobile phase ascended to the specified distance, the plates were removed from the chamber; solvent fronts were marked and allowed to dry at room temperature. The spots produced were observed under ultraviolet light. The color of each spot was noted and the
center of each spot was marked with a needle. The distance from the center of each spot to the point of application was measured and recorded (Anonymous, WHO 2002). Then the Rf value of each spot was calculated by using the formula. Rf = Distance between the point of application and the center of the spot / Distance between the point of application and the solvent front. RESULTS Rf value of Trikatu i.e., 0.68 is similar to standard Rf value of Shunthi i.e., 0.68. Rf value of Trikatu i.e., 0.09 is similar to standard Rf value of Maricha i.e., 0.09. Rf value of Trikatu i.e., 0.30 is similar to standard Rf value of Pippali i.e., 0.30. Hence it indicates the presence of Shunthi, Maricha and Pippali in Trikatu. Rf value of Hingwashtaka churna i.e., 0.65 is similar to standard Rf value of Trikatu i.e., 0.65. Rf value of Sutashekara rasa i.e., 0.18 is similar to standard Rf value of Trikatu i.e., 0.18. Hence it indicates that Trikatu is present in Hingwashtaka churna and Sutashekara rasa. (Table 1)
Table 1: Rf values under 366 nm UV light
Shunthi Maricha Pippali Trikatu Hingwashtaka churna Sutashekara rasa
a) 0.26 b) 0.64 c)0.68
a) 0.04
a) 0.03
b) 0.09
b) 0.10
c) 0.21
c) 0.16
a) 0.09 b) 0.18 c) 0.28
a) 0.03
a) 0.08
b) 0.21
b) 0.18
c) 0.65
c) 0.22
d) 0.30
d) 0.30
d) 0.58
e) 0.57
e) 0.65
e) 0.62
f) 0.63
f) 0.68 g) 0.71
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DISCUSSION As TLC is comparatively simple and rapid, the apparatus required is cheap and compound mixtures can be handled with comparative ease, hence is widely used in quality analysis of Ayurveda drugs. Standardization: The Rf values of Shunthi i.e., 0.26, 0.64 and 0.68 are standard values to that drug. Similarly Rf values of Maricha, Pippali, Trikatu are standard to those drugs. These standard values indicate the quality and purity of those drugs. Similarly TLC can be performed for other drugs and standards can be maintained which ensures the uniformity of thought and practice so that each and everyone can understand it in the same manner (Anonymous, PLIM 2003). So TLC is mentioned in monographs of Ayurveda drugs in the Ayurvedic Pharmacopoeia of India. Identification: TLC helps in proper identification of the right plant species. Single drug: Standards are selected as a model to which other test samples can be compared. Rf values of Shunthi, Maricha and Pippali are standard values. Any other market test samples of Shunthi, Maricha and Pippali can be compared with standard values. It should be similar to that of standard value, which indicates the presence of that drug. Compound drug: Trikatu contains a mixture of three drugs namely Shunthi, Maricha and Pippali. In order to know, whether these drugs were present in Trikatu, the Rf values of Trikatu were compared with the standard Rf
values of Shunthi, Maricha and Pippali. Rf values of Trikatu i.e., 0.68, 0.09, 0.30 are similar to standard Rf values of Shunthi, Maricha and Pippali i.e., 0.68, 0.09, 0.30 respectively. Hence it indicates the presence of Shunthi, Maricha and Pippali in Trikatu. Rf values may vary with each experiment depending on the saturation conditions in the chromatographic chamber, the activity of the adsorbent layer and the composition of the mobile phase. However, the analyst may use any other solvent system and detecting reagent if he is satisfied that the method which he uses, even by applying known reference standards, will give better result to establish the identity of any particular chemical constituent reported to be present in the drug (Anonymous, API 1999). CONCLUSION Market test samples of Hingwashtaka churna and Sutashekara rasa were selected to identify the presence of Trikatu in them. Rf values of Hingwashtaka churna and Sutashekara rasa are compared with standard Rf values of Trikatu. Rf value of Hingwashtaka churna i.e., 0.65 is similar to standard Rf value of Trikatu i.e., 0.65. Rf value of Sutashekara rasa i.e., 0.18 is similar to standard Rf value of Trikatu i.e., 0.18. Hence it indicates that Trikatu is present in Hingwashtaka churna and Sutashekara rasa. By this it can be confirmed that TLC can also be used to test the quality of market samples. Hence it can be concluded that Thin layer Chromatography is useful in Identification, Standardization and Qualitative evaluation of Ayurveda drugs.
REFERENCES Anonymous, (1999) - The Ayurvedic Pharmacopoeia of India, Government of India, Department of Ministry of Health and Family Welfare, the Control of Publication, Delhi, Part-I, Volume II.
Anonymous, (2003) - Phyto-chemistry, Standardization and Biotechnological aspects of ISM Drugs, PLIM publications, Ghaziabad, 15â&#x20AC;&#x201C;28.
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Anonymous, WHO, Geneva (2002) - Quality Control methods for medicinal plant materials, A.I.T.B.S. Publishers and Distributors (Regd.), 22–27.
Kirtikar K.R. & Basu B.D. (2005) – Indian Medicinal plants, International Book Distributors, Dehradun, Vol III, 2126– 2129.
Bhattacharjee A.K. & Das A.K. (1969) – Crude Drug Research, Vol IX, 1409.
Kokate C.K., Purohit A.P., Gokhar S.B. (2005) - Pharmacognosy, Nirali Prakashan, Pune, 67–86.
Chatwal and Anand (1996) - Instrumental methods of chemical Analysis, Himalaya publishing House, Bombay, 23–35. Chunekar K.C. & Pandey G.S. (2006) – Bhavaprakasha Nighantu of Sri Bhavamisra, Chowkambha Bharati Academy, Varanasi, 16–19.
Source of Support: Nil
Mishra
Shri Brahmashankar (2006) – Bhaisajyaratnavali of Shri Govinda Dasji, Chaukhambha Sanskrit Bhavan, Vol I, 643 & Vol III, 743.
Nadkarni K.M. (2007) - Indian Materia Medica, Vol I, Popular Prakashan Pvt.Ltd,Mumbai, Vol I, 965–969.
Conflict of Interest: None Declared
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 644–649 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Review article A REVIEW ON PHARMACODYNAMICS OF VAMANA KARMA AND VAMANOPAGA DASEMĀNI Ujjaliya Nitin1*, Remadevi R2 `1
Lecturer, Dept of Dravyaguna, Shri Dabur Dhanvantari Ayurveda College, Chandigarh, India Prof. & HOD. Dept of Dravyaguna, V.P.S.V. Ayurveda College, Kottakkal, Kerala, India *Corresponding Author: E-mail: drnujjaliya@gmail.com; Mobile: +918699327675
2
Received: 06/10/2012; Revised: 31/10/2012; Accepted: 04/11/2012
ABSTRACT Sodhana therapy is considered as the best management technique for a disease. Vamana karma is the first Pancakarma procedure and the best treatment for kapha dosha. Quick response of a drug on the body and the bodys’ response to the drug results Vamana karma. This procedure is quite difficult in some cases. Pharmacokinetics and Pharmacodynamics of Vamanopaga drugs is not clear yet. Pancakarma procedures are becoming popular among people as these cure diseases from the root cause. So it is necessary to establish mode of action of Vamana and Vamanopaga dravya for the betterment. This article is an effort to make a concept regarding mechanism of Vamana karma and Pharmacodynamics of Vamanopaga dasemāni of Caraka Samhitā. KEYWORDS: Vamana karma, Mechanism, Vamanopaga Dasemāni, Pharmacodynamics
To Cite this article: Ujjaliya Nitin, Remadevi R (2012), A REVIEW ON PHARMACODYNAMICS OF VAMANA KARMA AND VAMANOPAGA DASEMĀNI, Global J Res. Med. Plants & Indigen. Med., Volume 1(11), 644–649
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INTRODUCTION Vamana and Karma both words together make a procedure, Vamana karma (therapeutic emesis). This special procedure is preceded by Snehana (oleation) and Swedana (sudation) karmas. All together these procedures make a unique therapy for Kapha dosa. There are two types of conditions of dosas in the body. One is physiological, which maintain and helps in the functions of the body and the other is malabhūta (morbid), causing various diseases. Agnimāndya of Rasa and Raktadhātu produces morbid kapha and pitta respectively. The forcible expulsion of malarūpī kapha (morbid kapha) or excess pitta through the upper route is called Vamana. After preoperative procedures (snehana and swedana procedures) the process is followed by induction of Vamana by Vamana kalpas (medicine for Vamana), which are described in Chardanīya gana and Kalpa sthāna (Agnivesa, Caraka Samhita, 2001). The patient is advised to Samsarjana krama (specific dietary regimen) to reach up to normal diet and to complete the therapy. The whole procedure including Snehana, Swedana and Samsarjana krama is called Vamana karma. According to Ācarya Caraka, Virecana procedure, which is having the property of eliminating dosās from the body, is of two types. The removal of malarūpī dosās through oral route is called Vamana and through anal route is called Virecana (therapeutic purgation) (Agnivesa, Caraka Samhita, 2001). A lot of literature about indications and contraindications of Vamana karma are available in our classics. Selection of patient and Vamana kalpa (medicine) are the important factors for the procedure. Ācarya Caraka has mentioned six important drugs for Vamana and lots of combinations of them, in kalpa sthāna for different diseases (Agnivesa, Caraka Samhita, 2001). In the context of Mahākasāya, Ācarya mentioned Vamanopaga dasemāni (table no.1), ten drugs which are helpful in Vamana karma (Agnivesa, Caraka Samhita, 2003). Later Ācarya Vāgbhata mentioned Chardanīya gana having 21 drugs, Vamana and Vamanopaga (drugs help during
therapeutic emesis) together (Ācarya Vāgbhata, Astānga Hrudaya, 2003). Vāgbhata has not mentioned about upaga dravyas separately for vamana karma. Practically the drugs which are mentioned in kalpa sthāna are the main constitution of Vamana kalpas and drugs of Vamanopaga dasemāni are having supportive action on therapy. Definition and nirukti “Tatradosasharanam Urdhabhāgam vaman samjñakam” (Agnivesa, Caraka Samhita, 2001). ‘Vama dhatu’ adding by ‘ach’ pratyaya derived as word vamana. (Siddhānta Kaumudī) It means the purification of body or elimination of malarūpī dosas through oral route. Vamanopaga means helpful in Vamana and dasemāni means a group of ten drugs, which have similar pharmacological action, together or individual in case of Vamana karma. Mechanism of Vamana Karma Ācarya Caraka mentioned a deep conceptual mechanism of Vamana and Virecana karma. The difference between both mechanisms is in the mahābhaūtik composition of kalpa (medicine) thereby eliminating route. The mechanism described in the first chapter of kalpa sthāna said that, the drugs which are having Usna, Tiksna, Suksma, Vyavāyī, Vikāsī guna (hot, sharp, micro and fast acting properties), by their own Vīrya (potency) enters in to the heart. With the help of dhamanī (arteries) it enters in to both types of Srotas, Suksma and Sthūla (subtle and gross channels). After entering in to the all channels of the body due to Āgneya guna, it causes instant digestion and initiates the movement after softening. Here Tiksna guna separates the ready to go sticky dosās in the channels. After the detachment of dosa and mala from subtle and gross channels, these malarūpī dosās are ready to come in to mahāsrotas. A proper snehana and swedana procedure is helpful in this phenomenon. Ācarya has given a simile for this process. Due to oleation therapy, dosās will be not remaining in the Srotas. It is just like a well
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Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 644–649
oleated container never retains a drop of water inside it. Gunas (properties) of vamana drugs assist specific stages of vamana karma. Suksma property of medicine allows penetration into minute channels. Downward movement of dosās from suksma to mahāsrotas (gross channels) helps to come into Āmāsaya (Stomach). The pressure gradient developed by Anu-Pravanabāva (difference between subtle and gross channels) of medicine between Suksma and mahāsrotas helps to maintain a flow from channels. Due to the specialty of Vamana drugs (Agni and Vāyu mahābhūta dominant) (Maharsi Susruta, Susruta Samhita, 2005) these accumulated dosas and malas in the stomach move in the upward direction and gets expelled out, result Vamana karma (Agnivesa, Caraka Samhita, 2001). On the other hand this procedure facilitating the function of Udāna vāyu (one of the type of Vata dosa). Vāyu dosa is responsible for the movement of muscles involved in Vamana. Agni mahābhūta or Pitta dosa can be correlated with hormone or chemical trans-mediators of muscles and cells. Initiation of muscular contraction by Agni mahābhūta and Usnaguna, followed by Vāyu mahābhūta makes a forcible contraction of diaphragmatic and inter-costal muscles. Vyavāyī and Vikāsiguna make Vamana in faster pace. Due to the action of these gunas, drugs used in Vamana kalpa do not undergo digestion. All the drugs which possess emetic effect cannot be used for Vamana karma. The specialty of Vamana drugs is AnuPravanabāva, which is discussed above. Due to this, drugs are not deposited in the cells therefore not causing any complication. The Vamana drug goes to minute channels and returns quickly after exerting their effect at the site of action. This is the main difference between poisonous drug and Vamana drugs, though they have some similar property.
Observations during Vamana Karma 1. Perspiration – Liquidity of dosas in Srotas. 2. Romaharsa (horripilation) – Calatva (mobilization) of dosas. 3. Fullness of kostha (GIT) – Dosas have reached in to stomach. 4. Hrallās (nausea) – Dosas ready to get expelled out. Pharmacodynamic of drugs (Table no. 1) The overall Pharmacodynamic of Vamanopaga dasemāni drugs is based on guna concept. Most of the drugs (90%) are having property of Laghu and Ruksa guna. These are based on Vāyu, Agni and Ākasa mahābhaūtik (one of the five elements of the universe) composition. Ācarya Caraka has mentioned only the role of gunas in the Pharmacodynamic of Vamana karma (Bhadanta Nāgārjunā, Rasavaisesika, 2010). In fact guna is the thing which represents a drug. So, the selection of a drug should be on the basis of gunas for Vamana karma. (Table no. 2) Ācarya has mentioned predominance of Vāyu and Agni mahābhūta drugs for Vamana karma. Rasas (taste) of vamana dravyas are chiefly katu and kasāya rasa which are composition of the same mahābhūtas. Most of drugs are katu Vipāka having similar bhaūtic constitution. Other drugs are supportive to the therapy or to avoid complications during Vamana karma. As an example; honey which is mentioned in Vamanopaga dasemāni is added to Vamana kalpa (prepared medicine) for increasing the palatability and giving soothing effect. Āyurveda says it is a good kapha chedaka (expectorant), helps in better expulsion of malarūpī kapha by vamana karma. Likewise Saindhava (salt) should be added to Vamana kalpa for Vilāyana (Agnivesa, Caraka Samhita, 2001) (liquefying) of sticky Kaphadosa in channels. Effect of both the drugs is to help in a comfortable and irritation less procedure.
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 644–649
Table no. 1. Vamanopaga DaSemāni No. 01 02 03 04 05 06 07 08 09 10
Drugs Madhu Madhuka Kovidār Karbudār Nīp Vidul Bimbī Sanapuspī Sadāpuspī Pratyakapuspī
Latin name Honey Glycyrrhiza glabra. Linn. Bauhinia purpurea. Linn. Bohania variegata. Linn. Anthocephalus cadamba. Miq. Baringtonia acutangula. ( Linn) Gaemt. Coccinia indica. W & A. Crotalaria verrucosa. Linn. Calotropis procera. (Ait.) R. Br. Achyranthes aspera. Linn.
Table no. 2. Pharmacodynamic of drugs Rasa Katu Kasāya Tikta Madhura
Percentage 40 30 20 10
Guna Laghu, Ruksa Guru, Snigdha
Percentage 90 10
Table no. 3. Vamana and Vamanopaga Dravyas VAMANA DRAVYAS
VAMANOPAGA DRAVYAS
The main constitution of vamana kalpa (medicine) Single dose is adequate as per given in classics Vamana dravyas act in a faster pace with a minimum dose A therapeutic vamana kalpa should contain these drugs Vamana drugs act on the basis of Prabhāva
Supportive to the kalpa during therapeutic vamana Dosage can be changed according to Vegas during vamana Act under the influence of vamana drugs
Used according to disease
More than a disease depend on present condition of procedure Mainly act on mahāsrotas
Reach in all Srotas of the body
These can induce vomiting but cannot perform therapeutic vamana Act on the basis of Pancabhautik composition
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 644–649
In common practice hot infusion of Yasthīmadhu (Glycyrrhiza glabra Linn.) is used in higher dose for induction of better vega in vamana karma. Yasthīmadhu is sweet, demulcent, cool and soft in nature, in high dose acts as emetic but simultaneously soothes the throat. As per the Āyurvedic pharmacology it is a good kapha nihsāraka (expectorant) so helps in vamana karma for eliminating the dosa through oral route. Comparison of Vamana and Vamanopaga Drugs There is a modest difference between these drugs in their pharmacological actions because of their different bhaūtik composition and time of administration. (Table no. 3) Mechanism of Vomiting In the mechanism of Vomiting, different receptors are activated depending on the cause of vomiting or cause of nausea. For example 1.
Dopamine receptors in the chemotactic trigger zone (CTZ) in the fourth ventricle are stimulated by metabolic or drug causes of nausea.
2.
On the second way, gastric irritation stimulates histamine receptor in the vomiting centre via vagus nerve.
Vomiting is a highly integrated and complex reflex involving both autonomic and somatic neural pathway. Synchronous contraction of diaphragm, inter costal muscles and abdominal muscles raising the abdominal pressure, combined with relaxation of the lower esophageal sphincter, results in forcible ejection of gastric content. Stretch reflex and psychic stimuli are also the factors which initiate the sense of vomiting. (Davidson, Principles & Practice of Medicine, 2006) CONCLUSION Vamana drugs act on the basis of Usna, Tiksna, Suksma, Vyavāyī, Vikāsī gunas and bhaūtik composition. Drugs of Vamanopaga dasemāni support the whole procedure. Their action differs from drug to drug. From the modern point of view the drugs act via any of the pathways mentioned above. Thinking of individual guna and the overall effect of kalpa should be evaluated prior to the therapy. Vamanopaga daSemāni and Chardanīya gana of Susruta and Vāgabhata are the guidelines for making a suitable Vamana kalpa.
REFERENCE Agnivesa ’Caraka Samhita’, revised by Caraka and Drudhabala with ‘Ayurveda Dīpikā’ commentary, by Cakrapānīdatta, edited by Pt. Kashinath Sastri and Dr. Gorakhnath Chaturvedi (reprint 2001), Chaukhamba Vishvabharti, Varanasi -221001 (India), Kalpa Sthāna 1/4.
Agnivesa ’Caraka Samhita’, revised by Caraka and Drudhabala with ‘Ayurveda Dīpikā’ commentary, by Cakrapānīdatta, edited by Pt. Kashinath Sastri and Dr. Gorakhnath Chaturvedi (reprint 2003), Chaukhamba Vishvabharti, Varanasi -221001 (India), Sutra Sthāna 4/8/13.
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Global J Res. Med. Plants & Indigen. Med. | Volume 1, Issue 11 | November 2012 | 644–649
Astānga Hrudaya of Ācarya Vāgbhata edited by Prof. K.R. Srikantha Murthy (2003), Chaukhamba Krishnadas Academy, Varanasi, Sutra Sthāna 15 / 1.
Davidson Principles & Practice of Medicine, Sir Stanley Davidson, 20th Edition, Churchill Livingstone Elsevier Health Science, Philadelphia, (2006).
Bhadanta Nāgārjunā’s Rasavaisesika , edited by Vaidyabhushanam K. Raghvan Thirumulpad (2010,), Department of publication, Arya Vaidya Sala Kottakkal, 1st Edition Chapter 3 / 118124 & 3/45-51.
Susruta Samhita of Maharsi Susruta, edited by Āyurveda Tattva Sandīpikā by Kaviraj Ambikadatta Shastri (2005,), Chaukhamba Samskrit Samsthan, Varanasi -221001 (India), Sutra Sthāna 37/7.
Source of Support: Nil
Conflict of Interest: None Declared
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