GJRMI - Volume 5, Issue 7, July 2016

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INDEX – GJRMI - Volume 5, Issue 7, July 2016 MEDICINAL PLANT RESEARCH Ethno-Botany ETHNOMEDICINAL PLANTS USED FOR WOUND HEALING PURPOSES BY MALAYALI TRIBES OF KALRAYAN HILLS, SALEM DISTRICT, TAMIL NADU, INDIA M Kannan, T Senthil Kumar*, M V Rao

203–216

Bio-Chemistry EVALUATION OF THE HEPATOPROTECTIVE POTENTIAL OF HYDROETHANOLIC EXTRACT OF FICUS PUMILA L. ON CCl4 INDUCED LIVER DAMAGE IN RATS Christopher Larbie*, Dennis Torkornoo, Emmanuel Nyanor, Osei Asibey

217–225

Cover Page Photography: Dr. Hari Venkatesh K.R. Plant ID: Branch of Ingudi [Balanites aegyptiaca (L.) Delile]* of the family Zygophyllaceae; Place: Vriddhachalam, Cuddalore District, Tamil Nadu, India *Botanical Name validated from www.theplantlist.org as on 25/07/2016

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 203–216 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal

Research article ETHNOMEDICINAL PLANTS USED FOR WOUND HEALING PURPOSES BY MALAYALI TRIBES OF KALRAYAN HILLS, SALEM DISTRICT, TAMIL NADU, INDIA M Kannan1, T Senthil Kumar 2*, M V Rao3 1

Department of Botany, Directorate of Distance Education, Vinayaka Missions University, Salem-636 308, Tamil Nadu, India 2 Department of Industry University Collaboration, Bharathidasan University, Tiruchirappalli - 620 024, Tamil Nadu, India 3 Department of Plant Science, Bharathidasan University, Tiruchirappalli-620 024, Tamil Nadu, India *Corresponding Author: E-mail: senthil2551964@yahoo.co.in

Received: 27/05/2016; Revised: 19/07/2016; Accepted: 21/07/2016

ABSTRACT Ethnobotanical study plays an important role in the field of herbal research and enumeration of new medicinal plants. The tribal communities are dependent on the forest vegetation and utilize them for medicinal, non-medicinal, socio-cultural and religious purposes in their day-to-day life. The present paper documents the ethnomedicinal knowledge of Malayali tribes of Kalrayan hills on wound healing purposes only. Periodical field visits were made to collect the ethnomedicinal information on the plants used for wound healing purposes through personal interviews among the elders, village heads and traditional healers with the help of standardized questionnaires and discussions with them. Use of 72 plant species from 66 genera belonging to 43 families has been recorded along with the parts used, mode of preparation, mode of administration and their dosage. These plants are used in the form of paste, powder, juice, etc., for the treatment. In the present study, we observed that the Malayali tribes of Kalrayan hills are well experienced in the administration of local herbs for the treatment of wounds and other related injuries. KEYWORDS: Ethnobotany, Kalrayan hills, Malayali tribes, Traditional Healers, Medicinal Plants, Wound Healing, Salem.

Cite this article: M Kannan, T Senthil Kumar, M V Rao (2016), ETHNOMEDICINAL PLANTS USED FOR WOUND HEALING PURPOSES BY MALAYALI TRIBES OF KALRAYAN HILLS, SALEM DISTRICT, TAMIL NADU, INDIA, Global J Res. Med. Plants & Indigen. Med., Volume 5 (7): 203–216

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

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 203–216

INTRODUCTION Ethnobotany is a distinct branch of natural science dealing with the study of how people of a particular culture and region make use of indigenous plants for various aspects such as medicine, religion, cultural, agriculture instruments, house hold implements and several other disciplines (Patel and Patel, 2013). Now, ethnobotany has become a part of the tradition and culture through out the world. The uses of natural herbal drugs whether traditional or modern, have originated directly or indirectly from folklore and rituals known as ethnomedico-botany (Kutum et al., 2011). In the rural communities, wounds arising from bruises, cuts and scratches are untreated at the initial stages and such wounds became septic and inflamed before they are brought to the attention. In such cases, the wounds are treated with plant materials or it requires advice of herbalists (Grierson and Afolayan, 1999). A wound may be defined as a breakdown in the skin by the loss of continuity of epithelium with or without loss of underlying connective tissues such as muscles, bones, nerves, due to the injury to the skin or underlying tissues or organs caused by surgery, a blow, a cut, chemicals, heat, cold, friction, shear force, pressure or as a result of disease, such as ulcers. The most agreed phases of wound healing are inflammatory phase, proliferative phase and remodeling or maturing phase (Norman Williams et al., 2008). Wound healing occurs by regenerating dermal tissue as a natural process (Arshad Mehmood Abbasi et al., 2010). Throughout the world, wounds have been treated mostly topically with different medicinal herbs or with their extracts solely or in combination with some other plant parts (Ayyanar and Ignacimuthu, 2009). More than 70% of wound pharma products are plant based, 20% are mineral based and remaining contains animal products as their base material (Ramya Subramanian et al., 2011). There are several reports in adjacent hills of Kalrayan hills, around the Salem District of

Tamil Nadu, India on ethnomedicinal aspects (Sankaran and Alagesaboopathi 1995; Senthil Kumar and Krishnamurthy 1997; Alagesaboopathi 2009; 2011; 2012; 2013; 2014; Mishra et al., 2008; Parthipan et al., 2011; Rekha and Senthil Kumar, 2014). Though few workers have documented the ethnobotanical and ethnomedicinal plants in the study area, still lot of works are to be done (Kadavul and Dixit, 2009; Selvaraju et al., 2011; Natarajan et al., 2012; Manikandan and Alagu Lakshmanan, 2014; Kannan et al., 2015; 2016). Although most of the healers and elders are having vast knowledge on wound healing herbs, there is no separate report for wound healing plants in the study area. With this back ground, the present work was taken up to explore the ethnomedicinal plants used for the wound healing purposes and other related injuries such as cuts, burns, boils, sores, etc., by the Malayali tribes of Kalrayan hills, Salem district, Tamil Nadu, India. MATERIALS AND METHODS Study area Kalrayan (Kalvaryan) hills are a range of hills situated in the Eastern Ghats of the Southern Indian state of Tamil Nadu, lies between the North latitudes 11o 36‟ and 12o 01‟ N and the East longitudes 78o 29‟ and 78o 54‟ E. It runs over three districts viz. Salem (Southern and South Western portion), Villupuram (Central and Eastern portion) and a small region of northern part in Thiruvannamalai district and it stretches over an area of 1158.4 km2 (Sakthivel et al., 2010). The Kalrayan measures 25.76 km (NS) and 37 km (EW). The altitude varies from 1000 to 3800 meters above mean sea level (Natarajan et al., 2012). The average annual rainfall in the study area ranges from 782.98 to 1787.20 mm. The temperature varies from minimum of 25°C to a maximum of 40°C. It is composed of seven soil types and varies from red-loam to black clay (Kadavul and Parthasarathy, 2001) and is endowed with rich natural resources.

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

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 203–216

Divisions of Kalrayan hills Geographically, Kalrayan hills are divided into 5 regions or “Nadu”s (Cluster of tribal villages) namely, Chinnakalrayan Nadu (Northern part), Periyakalrayan Nadu (Western part), Jadaya Gaundan Nadu (Southern and Eastern part), Kurumba Gaundan Nadu (Central part) and Ariya Gaundan Nadu (North). Among them, Chinnakalrayan Nadu (Little Kalrayans) and Periyakalrayan Nadu (Big Kalrayans) belongs to Salem district and they include 58 and 44 tribal hamlets respectively. The remaining three regions belong to Villupuram district. Tribal community The tribal history of Kalrayan hills with its Jagirdars (Under the Muslim government, Jagirs are servants of the state, collect the revenue and looked the administration of the district/hills) dates back to the time of Krishna Deverayar, the Emperor of Vijaya Nagara Kingdom. The native people of the Kalrayans were called as Vedar (Hunter). The warriors belonging to Karalar community had invaded from Kanchipuram and settled in Kalrayan hills. After sometimes, they over rided the native people, Vedar of Kalrayan hills and married their girls. The mixed population of Karalar and Vedar communities presently called Malayali and they call themselves as Goundars (Velayutham Saravanan, 2003). Social status Ethnic people of Kalrayan hills are farmers and their life style is plain agricultural type. Due to the poor irrigational facilities, their agriculture is strictly unproductive, uneconomic and seasonal (Prabakaran et al., 2013). Most of them are poor and they are engaged in agriculture as Koolis. Only few people are in government services and some are engaged in cottage industries. To supplement their economy rest of the people are doing works interrelated with agriculture and involved in hunting, live stock keeping, poultry farming, rearing honeybees, collection of honey, bee wax and other minor forest products (Kuru

Suresh et al., 2011). In the off-seasons, (January – May months) the male members of the family travel to nearby towns such as Salem, Villupuram, Namakkal, Erode and Karur as daily wages or they migrate to the neighboring states like Karnataka or Kerala to compete their economy. They are interested in folktales, songs, worship, mythology, taboos, religious and social ceremonies (Thurston and Rangachari, 1909). Most of the tribal people of the study area are residing in remote and inaccessible forest areas and they are having no source to get modern medicine for their health care. Hence, they are practicing phytotherapy to treat the common ailments and they are having good knowledge on the herbal treatment for their health care. Enumeration of plants used for wound healing purposes is the part of the ethnobotanical research work carried out by us in the study area. Data Collection Ethnomedicinal data were collected through conversations and interviews with Vaidhyars (Traditional healers), tribal heads, elders and farmers having familiarity and sound knowledge on herbal treatments using standard methods adopted by Jain (1987). Regular field visits were made to the various hamlets of Kalrayan hills from Dec‟2009 to Dec‟ 2014 for the ethnomedicinal investigations. The medicinally important plants were collected, identified and verified with standard floras (Gamble, 1935; Mathew 1981-1983) and the voucher specimens were deposited in the Department of Botany, Vinayaka Missions University, Salem, Tamil Nadu, India. The documentation of ethnomedicinal information includes botanical name, local name, family, parts used, mode of preparation and administration. The information collected from the respondents of the study area were cross verified with the informants, healers or village headmen of the adjacent hamlets and also previously reported literatures of the study area and nearby hills.

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

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 203–216

RESULTS AND DISCUSSION

Wounds and related injuries

During the study period, we could able to understand that the large sections of tribal communities of the study area are well experienced in the administration of local herbs for various diseases. But this valuable herbal knowledge is confined to few members of the community in a hamlet. Most of the traditional healers learnt this knowledge from their ancestors from child hood by assisting them in the collection of herbal plants from wild, preparation of crude drugs, assisting in administration and other treatment procedures.

There are about 16 types of wounds and other related injuries noticed in the present study area (Table 1.) All these wounds fall under two major categories namely physical injuries and skin problems. About 11 types of injuries comes under the physical injuries and these type of wounds occur during various dayto-day life activities of the tribal community. Remaining five types of injuries occur as skin infections due to the microbial infections or as the impact of environmental factors.

Table 1: Various categories of wounds and other related injuries occurred among Malayali tribes of Kalrayan Hills, Salem, Tamil Nadu. S. No. 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16.

Name of the Diseases (Tamil) Kan Pungal Adi Paduthal Kuruma Pun (Naalpatta Pun) Mul Paduthal Padukkai Pun Periya Kaayam Pun Ratha Kattu Thee Pun Veekkam Vettu Kaayam Sirangu Koppalam Mukha Paru Kattigal Silanthi Kattigal

Name of the Diseases (English) Eye wounds / Injuries Injuries Wound Thorns in legs Bed sore Deep wounds Wound Contusion Burn wound Swelling Cut wounds Skin problems Boils Pimples Tumors Abscess

Ethnomedicinal plants For the treatment of wounds and other related injuries such as cuts, burns, boils, sores and wounds caused by external injuries, Malayali tribes of the study area are utilizing 72 plant species belonging to 66 genera under 43 families in which most of the plants are herbs (34 species), followed by tree (25 species), shrubs (7 species) and climbers (6 species). The predominant families are Apocynaceae with 5 species, Euphorbiaceae,

Terminology

Category

Ocular trauma Physical injuries Trauma Chronic wound Wound Decubitus ulcer Wound Wound Haematoma Burns Inflammation Laceration Impetigo Skin problems Furuncle / Boils Acne Neoplasm Abscess

Fabaceae and Rubiaceae with 4 species each followed by Capparaceae, Malvaceae, Moraceae, Rutaceae and Solanaceae with 3 species each. Remaining families are represented by one or two species each (Table 2). The plants were enumerated alphabetically under each disease along with the information such as their botanical name, vernacular name, family, part of the plant used and mode of preparation and administration (Table 3).

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

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 203–216

Bryophyllum pinnatum, Cassia fistula, Clausena dentata, Calotropis gigantea, Erythrina indica, Jatropha curcas, Lantana camara, Pterocarpus marsupium, Ricinus communis and Tridax procumbens are some of

the notable plants used for the wound healing purposes and it is in agreement with previous reports (Ramya Subramanian et al., 2011; Shrirame and Gogle, 2014; Ramesh Kumar Ahirwar and Kumud Sandya, 2015).

Table 2: Family wise distribution of ethnomedicinal plants used for wound healing purposes in Kalrayan hills, Salem district. S. No.

Family name

No. of plant species (in each family) 5

1.

Apocynaceae

2.

Euphorbiaceae, Fabaceae, Rubiaceae

4

3.

Capparaceae, Malvaceae, Moraceae, Rutaceae, Solanaceae

3

4.

Asteraceae, Boraginaceae, Caesalpiniaceae, Convolvulaceae, Liliaceae, Mimosaceae Acanthaceae, Aizoaceae, Amaranthaceae, Apiaceae, Arecaceae, Cactaceae, Caricaceae, Celastraceae, Crassulaceae, Dioscoriaceae, Ebenaceae, Hyacinthaceace, Hypoxidaceae, Lamiaceae, Legoniaceae, Melastomataceae, Musaceae, Nyctaginaceae, Oxalidaceae, Pedaliaceae, Piperaceae, Poaceae, Polygonaceae, Pteridaceae, Salvadoraceae, Sapotaceae, Verbenaceae, Zingiberaceae

2

5.

1

Table 3: List of Ethnomedicinal plants used for wound healing purposes by Malayali Tribes of Kalrayan Hills, Salem, Tamil Nadu. S. No.

Diseases

Binomial Name

Vernacular Name

Family

Parts used

Formulations and doses

1.

Kan Pungal (Eye Wounds)

Clausena dentata M. Roem.

Nana chedi

Rutaceae

Leaf

Diospyros Roxb. Tephrosia (L.) Pers.

Vakana Maram Oosi Thuvarai

Ebenaceae

Leaf

Young leaves ground with mother milk and extract is applied externally in eyes as drops. Leaf juice used as eye drops.

Fabaceae

Leaf

Palai

Apocynaceae

Leaf

Vel Velam

Mimosaceae

Bark

Nayuruvi

Amaranthaceae

Leaf, Stem

Wrightia R.Br.

2.

Pungal (Wound Healing)

montana purpurea

tinctoria

Acacia leucophloea Willd. Achyranthes aspera L.

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

Two or three drops of leaf juice is administered as eye drops to cure wounds made by stick injuries. Also used in cattle for the same purpose. Crushed leaf extract mixed with mother's milk. Clean white cloth is dipped in the extract and administered as eye drops in nights. Bark paste in hot water is applied externally. Paste is applied externally.

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 203–216 Biophytum sensitivum (L.) DC. Blepharis maderaspatensis B.Heyne ex Roth

Male surungi

Oxalidaceae

Leaf

Leaf paste is applied externally.

Nenthria Poondu

Acanthaceae

Leaf

Bryophyllum pinnatum (Lam.) Kurz

Ranakalli

Crassulaceae

Leaf

Leaf juice with Sesamum indicum oil is boiled and applied on the wounds twice or thrice a day. Leaf paste is applied on wounds, insect bites and boils to reduce the pain and severity.

Cleome gynandra L.

Velai Chedi

Capparaceae

Root

Cleome viscosa L.

Naai velai

Capparaceae

Leaf

Cleome viscosa L.

Naai velai

Capparaceae

Leaf

Dioscorea bulbifera L.

Kaattu Vethala Kodi

Dioscoreaceae

Tuber

Erythrina indica Lam.

Kalyana Murungai

Fabaceae

Leaf

Euphorbia heterophylla L. Ficus microcarpa L.f.

Paal Perukki

Euphorbiaceae

Leaf

Kal Arasan

Moraceae

Bark, Leaf

Heliotropium indicum L. Aloe vera (L.) Burm.f.

Boraginaceae

Seed

Liliaceae

Leaf

Opuntia dillenii Haw.

Thel Kodukku Sotru Katralai Sappathikalli

Cactaceae

Fleshy

Calotropis procera (Aiton) W.T.Aiton Cocos nucifera L.

Vellai Erukan Thennai

Apocynaceae

stem Leaf

Arecaceae

Oil

Ipomoea asarifolia Roem. & Schult. Pavonia zeylanica (L.) Cav. Centella asiatica (L.) Urb. Ixora coccinea L.

Sundan chedi kodi Vedivana poondu Vallarai

Convolvulaceae

Leaf

Malvaceae

Leaf

Apiaceae

Leaf

Idli Poo

Rubiaceae

Leaf,

It is specially used to heal the wounds made by broken glass pieces. Root paste is applied and tied on the wound area for a day. The broken glass pieces will be dissolved and oozes out if any inside the wound. Leaf paste is applied externally up to cure. Leaf juice is applied externally up to cure. Tuber is ground well and applied on the wounds externally. Leaf juice is applied externally to stop bleeding and leaf paste tied on the wounds to heal. Leaf paste is applied externally. Bark paste is used for healing wounds and leaf decoction also used for cleaning and washing the wounds. Seed paste is applied externally. Along with plant materials small quantity of sugar is added and ground well in to paste and applied externally. Dried leaf powder is mixed with coconut oil and applied externally as paste. Leaf paste is applied externally up to cure.

Dry decoction is externally on wounds.

applied

Flower

Madhuca longifolia J.F.Macbr. Mirabilis jalapa L. Mitragyna Korth.

parvifolia

Iluppai

Sapotaceae

Bark

Anthi Mantharai Kadamba Maram

Nyctaginaceae

Root

Rubiaceae

Leaf

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Root paste or powder is applied externally up to cure. Leaf paste is used to dress wounds.

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 203–216

3.

4.

5.

6.

Vettu Kaayangal (Cut Wounds)

Periya Kaayangal (Deep Wounds) Kuruma Pun (Naalpatta pun Chronic Wounds)

Thee

Pun

Musa paradisiaca L.

Vazhai

Musaceae

Latex

Opuntia dillenii Haw. Strychnos nux-vomica L. Sesamum indicum L. Waltheria indica L.

Sappathikalli Yetti Maram

Cactaceae Loganiaceae

Fruit Seed

Ellu Sengali Poondu Sangamullu

Pedaliaceae Malvaceae

Oil Root

Salvadoraceae

Root

Mavilangam

Apocynaceaee

Root

Unnichedi

Verbenaceae

Leaf

Premna tomentosa Willd. Tridax procumbens L.

Poda nari

Lamiaceae

Leaf

Vettu Kaya Poondu

Asteraceae

Leaf

Biophytum sensitivum (L.) DC. Canthium parviflorum Lam. Curculigo orchioides Gaertn. Naringi crenulata (Roxb.) Nicolson Curcuma longa L.

Male surungi

Oxalidaceae

Leaf

Karai

Rubiaceae

Leaf

Nilapanai kizhangu Pori valai Poondu Manjal

Hypoxidaceae

Rhizome

Rutaceae

Leaf

Zingiberaceae

Sida rhombifolia L.

Marunthu Thalai

Malvaceae

Rhizo -me Leaf

Curcuma longa L.

Manjal

Zingiberaceae

Rhizome

Dendrocalamus strictus Nees Calotropis gigantea (L.) W.T. Aiton

Siruvarai Mungil Erukku

Poaceae

Leaf

Apocynaceae

Latex

Latex is applied on the wounds two or three times daily up to cure.

Allium sativum L. Ipomoea asarifolia Roem. & Schult. Pavonia zeylanica (L.) Cav. Piper nigrum L. Hemionitis cordifolia Roxb. in Wall.

Poondu Sundanchedi kodi Vedivana poondu Milahu Kal paruthi

Liliaceae Convolvulaceae

Bulb Leaf

Malvaceae

Leaf

Plant materials are ground well with Allium sativum and Piper nigrum and paste is applied externally up to cure.

Piperaceae Pteridaceae

Seeds Leaf

Cassia fistula L.

Sara kondrai

Caesalpiniaceae

Leaf

Azima tetracantha Lam. Decalepis hamiltonii Wight & Arn. Lantana camara L.

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Latex is applied on the wounds and dressed with clean white cloth. Then 2 or 3 drops added on the dressing. Cloth is persistent on wounds up to cure. Fruit pulp is applied externally. Paste is applied externally. Dry powder is mixed with Sesamum oil and applied externally. Paste is applied externally. Root powder is applied externally to cure cut wounds. Paste is applied externally. As it dries up and persistent on the wounds up to cure, it may form pus in some persons. Leaf paste is roasted and applied externally. Leaf juice will control the bleeding immediately and paste is applied externally on wounds to heal. Young leaves are made in to paste and applied externally up to cure.

Leaf is ground with fresh or dry turmeric and made in to paste. It is applied externally on wounds and this paste is persistent on wounds up to cure. Leaf paste is applied externally on cut wounds with Curcuma longa powder.

Ground with cow ghee and applied externally up to cure. Maantham, Selanthi Kattikal, wounds in knuckles of fingers are also cured. Fresh leaves are ground with

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 203–216 (Burns)

Pungal & Muka Paru (Wounds & Pimbles) Sirangu (Impetigo)

7.

8.

Koppalam (Furuncle / Boils)

9.

Cocos nucifera L.

Thennai

Arecaceae

Oil

Hemionitis cordifolia Roxb. Wall. Musa paradisiaca L.

Kalparuthi

Pteridaceae

Leaf

Vazhai

Musaceae

Leaf

Polygonum glabrum Willd. Memecylon umbellatum Burm.f.

Attarali

Polygonaceae

Leaf

Allan Maram

Melastomataceae

Leaf

Acacia concinna DC.

Seevakkaai

Mimosaceae

Fruit

Acalypha indica L.

Kuppai Meni

Euphorbiaceae

Leaf

Cordia myxa L.

Naruvili

Boraginaceae

Bark

Ipomoea quamoclit L.

Mayil Manikkam Therani chedi

Convovulaceae

Leaf

Rubiaceae

Leaf

Citrus limon (L.) Burm.f. Mollugo nudicaulis Lam. Xanthium indicum J.Koenig ex Roxb. Hemionitis cordifolia Roxb. Wall.

Elumichai

Rutaceae

Fruit

Parpadakam

Aizoaceae

Leaf

Dry powder is used as antiseptic for washing wounds. Paste is applied externally up to cure. Stem bark paste is externally applied up to cure. Crushed leaves are applied externally on boils up to cure. Leaf paste is applied on the boils and boils with pus and also applied externally on skin allergies. Leaf is ground with lime juice and applied externally up to cure.

Otta Chedi

Asteraceae

Leaf

Leaf juice is applied externally.

Kal paruthi

Pteridaceae

Leaf

Tarenna Kuntze

Therani chedi

Rubiaceae

Leaf

Cleome monophylla L.

Ellu Sakkalathi

Capparaceae

Leaf, Seed

Wattakaka volubilis Stapf Pterocarpus marsupium Roxb.

Peria Kurinjan Vengai Maram

Apocynaceae

Leaf

Fabaceae

Leaf

Leaves are ground with cow ghee and applied on the infected regions. Leaf paste is applied on the boils and boils with pus and also applied externally on skin allergies. Either seed or leaf of the plant is ground and applied on the boils or cut wounds. It will cure and prevent the formation of pus. Leaf paste is applied externally on the surfaces. Crushed leaves are applied externally on boils and sores.

Azima tetracantha Lam. Carica papaya L.

Sangamullu

Salvadoraceae

Root

Pappali

Caricaceae

Latex

Urginea indica Kunth

Kaatu Poondu

Hyacinthaceae

Bulb

Withania (L.) Dunal

Amukara Kizhangu

Solanaceae

Tuber

Tarenna Kuntze

10.

11.

12.

Furuncle between Fingers Boils & Skin Allergies Boils Wounds

13.

Boils Sores

14.

Kattigal (Tumors)

&

&

asiatica

asiatica

somnifera

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coconut oil and made in to paste and applied externally. Leaf paste is applied externally. In heavy burns the persons will be laid down in the leaves up to the recovery. Leaf paste is applied externally for burn wounds. Leaf paste is applied externally up to cure.

Root is ground with water and applied externally. Latex is applied on the tumors daily up to cure. Ground and paste is applied externally on tumors for 2 or 3 days. Paste is applied on the surfaces will control the Kattihal (tumers), Amukara means amukuthal meaning suppress the tumors.

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 203–216 15.

Silanthi Katti

Lycopersicon esculentum Mill.

Thakkali

Solanaceae

Fruit

16.

Adi Paduthal & Veekkam (Trauma & Swelling) Swelling & Boils

Artocarpus heterophyllus Lam. Cassine glauca (Rottb.) Kuntze

Pala Maram

Moraceae

Latex

Karuvaali Maram

Celastraceae

Root

Datura metel L.

Karuoomath ai

Solanaceae

Leaf

Leaf is ground in to paste with hot water and applied externally.

Tamarindus indica L.

Puli

Caesalpiniaceae

Leaf

Erythrina indica Lam.

Kalyana Murungai Kattu Kottai

Fabaceae

Leaf

Euphorbiaceae

Leaf

Poda nari

Lamiaceae

Leaf

Kottai Maram Kaatu Pala

Euphorbiaceae

Oil

Moraceae

Bark

Phaseolus mungo L.

Ulundhu

Fabaceae

Seed

Sesamum indicum L.

Ellu

Pedaliaceae

Oil

Premna Willd.

Poda nari

Lamiaceae

Leaf

Sesamum indicum L.

Ellu

Pedaliaceae

Oil

Calotropis gigantea (L.) W.T.Aiton

Erukku

Apocynaceae

Latex

Leaf is ground in to paste with hot water and applied externally. Also used to treat pain. Leaf paste is applied externally on the surfaces. Leaves are soaked in rice water overnight, heated and tied around painful area to reduce pain. Leaves are boiled in water and fixed on the injured area by means of castor oil. It will be continued up to cure. Fresh or dried bark powder and Phaseolus mungo seed powder are mixed together in equal quantities. Sesamum oil and egg white yolk are mixed with the above powder and made in to paste. Applied externally on surfaces of the injured regions, bone fractures and Ratha kattu (blood clotting in injured area). Sesamum oil is applied on the leaf surface of Premna tomentosa and shown in direct fire for few seconds (Vaatti) and they are applied on the injured or fractured area. For removal of thorns from the leg, a drop of latex is applied on the thorny area. On the next day thorns can be removed easily. It also heals the wounds formed due to the thorns.

17.

18.

Ratha Kattu (Contusion)

Jatropha curcas L.

Premna tomentosa Willd. Ricinus communis L. 19.

20.

Ratha Kattu,Bone fracture, Injuries

Thorns legs

in

Artocarpus Lam.

hirsutus

tomentosa

Parts of the plants used For wound healing treatments, plant parts such as leaf, stem, root, bark, latex, seed, flower, fruit, tuber, bulb, rhizome and seed oil are used by the local healers and elders of the study area. Leaves were found to be most frequently used for the treatment, which accounts for 52 preparations followed by root (Seven preparations) and oil (Six preparations).

Fruit is made in to small pieces and tied with white cloth on the surface of the Silanthi Katti for 3 to 4 days. Latex is applied externally daily up to cure. Root paste is applied externally.

Other plant parts are represented in few preparations only (Figure 1). Tribes of Kalrayan hills are using different dosage forms for wound healing purposes. Among them, use of fresh plant parts as such, dry powder, pastes, juice, decoction, combinations of various parts of plants are some of the important dosage forms used by them. There are about 74 preparations in use

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among the villagers in which use of paste form is dominating over all other dosage forms with 47 preparations (Figure 2). Jatropha curcas and Premna tomentosa leaves are processed with oil and shown in fire directly then applied on the wounds for the healing process. Latex from Artocarpus heterophyllus and Carica papaya are also used for the healing purposes.

In certain cases, two types of preparations of the same plant were used in a single treatment. For example, Ficus microcapa leaf decoction is used for cleaning the wounds where as, its bark paste is applied externally on wounds for healing purposes. In the same way, leaf juice of Tridax procumbens is used for the control of bleeding in cut wounds where as its paste is applied externally to heal the wounds.

Figure 1: Various parts of ethnomedicinal plants used by Malayali tribes of Kalrayan hills for wound healing purposes. Leaf 3

Root

21 2 2

Oil

4

Bark

5

Latex Seed

5

Fruit

5

Rhizome

52

Stem 6

Tuber Bulb

7

Flower

Figure 2: Various modes of preparations of medicaments used for the wound healing purposes by Malayali tribes of Kalrayan hills, Salem.

3

2 2 Paste

4

Juice

6

Fresh Parts Powder Processed Materials

10 47

Decoction Latex

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Modes of administration Maximum number of preparations used for promoting wound healing was used as external application (68) and only two preparations are administered orally. Many researchers have reported the external application as the dominant mode of administration in wound healing treatments (Senthil Kumar and Krishnamurthy, 1997; Grierson and Afolayan, 1999; Ayyanar and Ignacimuthu, 2009; Ramya Subramanian et al., 2011). As the treatment for wound healing is continued up to cure, dosage of administration is not recommended for any of the preparations. Bandages made using the bark extract of Musa paradisiaca and pastes prepared from the plants such as Curcuma longa, Lantana camara, Sida rhombifolia etc., when applied are persistent on the wounds till it heals and the dried bandages and plant pastes will fall off automatically after the healing. As the tribal communities of the study area are forest dwellers, they depend on their surrounding for their day-to-day activities. It was observed that the wounds are one of the major problems suffered by those forest inhabitants due to their life style in the forest area and they easily get injured during their day-to-day activities in the forest. As they are dwelling and working in the interior regions of the forests, immediate need for the remedy arises for their wounds with in the forest and they go for phytotherapy. Hence, most of them learnt the herbal knowledge from their mother, father, elders of the family, neighbors, and relatives etc., who are nearer to them at the time of injuries in the forest. CONCLUSION The present documentation revealed that the ethnomedicinal plants are still in use among Malayali tribal communities of the study area and they are well experienced in the administration of local herbs for the treatment of wounds and other related injuries. These plants are common, cost effective and easily accessible at the time of requirement. But the

practice of herbal medicine is being utilized by few „Vaidhyars‟ and elderly people only. It is concluded that the healthcare system of Malayali tribe traditionally depends on herbal practices for their ailments. Moreover, elders and traditional healers have rich knowledge on herbal wealth. The traditional knowledge of the healers helped us in the documentation of the ethnobotanical information. Hence, the documentation of traditional knowledge will help in the field of herbal research and enumeration of new medicinal plants for a particular therapeutic effect. At the same time the detailed pharmacological investigations of the herbal plants will be helpful in development of newer drugs for a particular condition. ACKNOWLEDGEMENTS One of the authors (MK) is thankful to The Chancellor, Vinayaka Missions University, Salem. The authors are also thankful to the resident tribes of the study area for their response and participation in the survey by sharing their knowledge on plants. The authors are also grateful to Dr. R. Prabakaran, Vivekanandha College of Arts and Sciences (W), Tiruchengode for accompanying us during the field visit. A special word of gratitude for tribal informant Mr. A. Murugesan who helped the author a lot during the course of study and Dr. A. Anuradha, Department of Obstetrics & Gynecology, Sivaraj Homeopathy Medical College & Research Institute, Salem for the suggestions.

REFERENCES Alagesaboopathi C (2009). Ethnomedicinal plants and their utilization by villagers in Kumaragiri Hills of Salem district of Tamilnadu, India. Afr J Trad Complement Altrn Med 6(3):222–227. Alagesaboopathi C (2011). Ethnobotanical studies on useful plants of Kanjamalai Hills of Salem district of Tamil Nadu, Southern India. Arch Appl Sci Res 3(5):532–539.

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Alagesaboopathi C (2012). Ethnobotanical survey of medicinal plants used by Malayali tribals and rural people in Salem district of Tamilnadu, India. J Pharm Res 5(12):5248–5252

Kadavul K, Dixit AK (2009). Ethnomedicinal studies of the woody species of Kalrayan and Shervarayan Hills, Eastern Ghats, Tamil Nadu. Indian J Tradit know 8(4):592–597.

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Kannan M, Senthil Kumar T, Rao MV (2016). Utilization of plant resources for nonmedicinal purposes by Malayali tribes of Kalrayan hills, Salem district, Tamil Nadu, India, International Journal of Herbal Medicine, 4(1): 47–57.

Ayyanar M, Ignacimuthu S (2009). Herbal medicines for wound healing among tribal people in Southern India; Ethnobotanical and Scientific evidences, International Journal of Applied Research in Natural Products, 2(3):pp.29–42. Gamble JS, Fischer CEC (1935). Flora of the Presidency of Madras, London, Adlard and Son, Ltd., Calcutta, Vol I-III. Grierson DS, Afolayan AJ (1999). An ethnobotanical study of plants used for the treatment of wounds in the Eastern Cape, South Africa. Journal of ethnopharmacol, 67:327–332. Jain SK (1987). Ethnobotany, its scope and various sub-disciplines, In: A manual of Ethnobotany. S.K. (Edu). Scientific publishers Jodhpur, pp1–11.

Kuru Suresh, Kottaimuthu R, Selvin Jebaraj Norman T, Kumuthakalavalli R, Sabu M Simon (2011). Ethnobotanical study of medicinal plants used by Malayali tribals in Kolli hills of Tamilnadu, India, International Journal of Research in Ayurveda & Pharmacy, 2(2):502– 508. Kutum A, Sarmah R, Hazarika D (2011). An ethnobotanical study of Mishing tribe living in Fringe villages of Kaziranga national park of Assam, India, Indian Journal of Fundamental and Applied Life Sciences, 1(4): pp.45–61 Manikandan S, Alagu Lakshmanan GM (2014). Ethnobotanical survey of Medicinal Plants in Kalrayan hills, Eastern Ghats, Tamil Nadu. Int Lett Nat Sci, 12(2): 111–121. Matthew KM (1983). The Flora of Tamil Nadu Carnatic (The Rapinat Herbarium, St. Joseph‟s College, Tiruchirapalli, India), Vol I-III.

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Mishra SB, Dwivedi S, Shashi A, Prajapati K (2008). Ethnomedicinal uses of some plant species by ethnic and rural people of the Salem district of Tamilnadu with special reference to the conservation of vanishing species. Ethnobot Leaflet, 12: 873–887. Natarajan V, Anbazhagan M, Rajendran R (2012). Studies on Ethnomedicinal plants used by the Malayali tribe of Kalrayan Hill, Tamil Nadu state. Res Plant Biol, 2(1):15–21. Norman S Williams, Christopher JK, Bulstrode, P. Ronan O‟ Connell (2008). Bailey & Love‟s Short practice of surgery, 25th edition, Hodder Arnold, London, pp24–26. Parthipan M, Aravindhan V, Rajendran A (2011). Medico-botanical study of Yercaud Hills in the Eastern Ghats of Tamil Nadu, India. Ancient Sci Life, 30(4):104–109. Patel

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Prabakaran R, Senthil Kumar T, Rao MV (2013). Ethnoforestry and ethnoagricultural knowledge of Malayali tribes of Chitteri hills, Tamil Nadu, Journal of Biodiversity and Environmental Sciences, 3(5):12–19. Ramesh Kumar Ahirwar, Kumud Sandya (2015). Ethnobotany and Ethnoveterinary Plants used for Wounds Healing by Baiga Tribes Umaria District, Madhya Pradesh, India, IJPPR. Human, 4(1):159–166.

Ramya Subramanian, Gopinath Krishnasamy, Aruna Devaraj, Padmavathy Sethuraman, Ramaraj Jayakumararaj (2011). Wound healing ethnopharmocological potentials of selected medicinal plants used by Malayali tribes, International Research Journal of Pharmacy, 2(5):132–137. Rekha

R, Senthil Kumar S (2014). Ethnobotanical plants used by the Malayali tribes in Yercaud hills of Eastern Ghats, Salem District, Tamil Nadu, India. Global J Res Med Plants Indigen Med, 3(6):243–251.

Sakthivel R, Manivel M, Jawahar Raj N, Pugalanthi V, Ravichandran N, Vijay D. Anand (2010). Remote sensing and GIS based forest cover change detection study in Kalrayan hills, Tamil Nadu. J Environ Biol, 31(5):737–747. Sankaran S, Alagesaboopathi C (1995). Some medicinal plants used by the tribals of Shevaroy Hills, Tamilnadu. Int J Flora Fauna, 1:137–138. Selvaraju A, Ayyanar M, Rathinakumar SS, Sekar T (2011). Plants used in ethnoveterinary by Malayali tribals in Salem district of Tamil Nadu, India. Med plants, 3(3):1–7. Senthil Kumar T, Krishnamurthy KV (1997). Ethnobotanical study on Shevaroy hills of Eastern Ghats. Solai Bull Ethnopharmacol, pp.31–36. Shrirame AM, Gogle DP (2014). Survey on ethnobotanical plants used for wound healing:Nagpur region, Int J of Life Sciences, Special issue A2:62–64. Thurston E, Rangachari K (1909). Castes and Tribes of Southern India, Government press (Reprinted 1975), Cosmo puplication, New Delhi, pp.406–436.

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Velayutham Saravanan (2003). Colonial commercial forest policy and tribal private forests in Madras Presidency: 1792–1881, Indian Economic Social History Review, 40:403. Source of Support: NIL

Conflict of Interest: None Declared

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 217–225 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal

Research article EVALUATION OF THE HEPATOPROTECTIVE POTENTIAL OF HYDROETHANOLIC EXTRACT OF FICUS PUMILA L. ON CCl4 INDUCED LIVER DAMAGE IN RATS Christopher Larbie1*, Dennis Torkornoo2, Emmanuel Nyanor3, Osei Asibey4 1,2,3

Department of Biochemistry and Biotechnology, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana 4 Biochemistry Laboratory, KomfoAnokye Teaching Hospital, Kumasi, Ghana *Corresponding Author: E-mail: clarbie.cos@knust.edu.gh

Received: 27/05/2016; Revised: 19/07/2016; Accepted: 21/07/2016

ABSTRACT Liver diseases and defects are on the constant rise all around the world, with the available pharmaceutical drugs not being efficient for treatment and also presenting adverse effects after use. The general objective of the study was to determine the hepatoprotective effect of 50% hydroethanolic extract of Ficus pumila on carbon tetrachloride (CCl4) induced liver damage in rats. In this work, liver damage was induced in experimental animals by administering 1 ml/kg CCl4 orally, after which the rats were treated with 50% hydro-ethanolic extract of F. pumila and Silymarin, a standard hepatoprotective drug. 50% hydroethanolic extract at 100 and 250 mg/kg were administered for 7 days by oral route. Hepatoprotective effect was studied by assaying the activity of hepatobiliary enzymes such as ALT, AST, ALP, GGT and biochemical parameters such as cholesterol, triglycerides, high density lipoproteins, total bilirubin, direct bilirubin concentration and indirect bilirubin concentration. Body weight and liver weights (absolute and relative) of the rats were also measured. The activity of all the hepatobiliary enzymes registered significant increases upon CCl4 administration. There were also increases in cholesterol and triglyceride concentration and in total and indirect bilirubin, an indication of liver damage. F. pumila at all doses significantly (0.05– 0.001) restored liver function compared to the Silymarin control and also increased body and absolute liver weight. The results therefore indicate that leaves of F. Pumila possess hepatoprotective activity. KEYWORDS: Hepatoprotective, Ficus pumila, carbon tetrachloride, Silymarin, liver damage.

Cite this article: Christopher Larbie, Dennis Torkornoo, Emmanuel Nyanor, Osei Asibey (2016), EVALUATION OF THE HEPATOPROTECTIVE POTENTIAL OF HYDROETHANOLIC EXTRACT OF FICUS PUMILA L. ON CCl4 INDUCED LIVER DAMAGE IN RATS, Global J Res. Med. Plants & Indigen. Med., Volume 5 (7): 217–225

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INTRODUCTION ‘Liver diseases’ is a broad term that covers all the potential problems that leads to the failure of the liver to perform designated functions (MedicineNet.com, 2015). The liver’s fundamental role in human physiology includes metabolism of nutrients (carbohydrates, lipids and proteins) and excretion of drugs and other xenobiotics (Abraham, 2014). According to Friedman et al. (2003), more than 900 drugs have been implicated in causing liver injury, which leads the chart of reasons why pharmaceuticals are removed from the market. Hepatotoxicity and drug-induced liver injury also account for a substantial number of compound failures, highlighting the need for drug screening assays such as stem cell-derived hepatocyte-like cells that are capable of detecting toxicity early in the drug development process (Greenhough and Hay, 2012). The deleterious usage of these chemicals usually causes subclinical injury to the liver, which manifests only as abnormal liver enzyme tests. Hepatotoxicity is responsible for 5% of all hospital admissions and 50% of all acute liver failures (McNally, 2006; Ostapowicz et al., 2002). Upon hepatotoxic exposure, the patients that end up with drug-induced liver defects add on to the already existing outrageous prevalence of liver diseases. The percentage of people affected by liver damage is constantly rising all around the globe. In the year 2003, established cirrhosis had a 10-year mortality of 34–66% largely dependent on the cause of the cirrhosis; alcoholic cirrhosis had a worse prognosis than primary biliary cirrhosis and cirrhosis due to hepatitis. The risk of death due to all causes was increased twelve fold. If one excluded the direct consequences of the liver disease, there was still a five-fold increased risk of death in all disease categories (Sørensen et al., 2003). Although all causes of these liver defects aside drug induced hepatotoxicity are not thoroughly known, certain investigations and studies bring out most probable causes which eventually get accepted, at least theoretically, as causes.

In order to overcome the effect of hepatotoxicity caused by certain pharmaceuticals, interest has been shifted towards the use of alternative medicines. Traditional medicines and extracts from medicinal plants have been extensively used as alternative medicine for better control and management of certain diseases. Plants have always been an essential part of human civilization. Many plants and herbs are naturally scattered all over the face of the earth. Each one of them possesses various significant properties and qualities that enable it to be used as medicine for the treatment of various ailments (Ahmed et al., 2013). Numerous studies have been conducted on the composition of Ficus pumila, resulting in the identification of a number of compounds as constituents. These include apigenin and luteolin (Harborne, 1986) and also rutin, genistein, hesperidin, astragalin, isoquercitrin and chrysin (Morelli et al., 2000). The Ficus pumila L. (family: Moraceae) is an ornamental plant usually cultivated for its aesthetic properties with many great properties and as a result has been used by different people worldwide for positive health purposes. It is prepared either as a beverage or herbal medicine to treat diabetes, high blood pressure, dizziness and neuralgia by the Okinawan folks in Japan (Mitsuhashi, 1988). The leaves of Ficus pumila serve as a good and rich natural source of various antioxidants and also phenolic acid compounds; flavanoid glycosides and antioxidant properties have been reported from the 50% hydroethanolic leaf extract of Ficus pumila (Leong et al., 2008; Larbie et al., 2015). Further, tannins, saponins, general glycosides, alkaloids and triterpenes have been identified in the leaf and stem extract with leaves showing high levels of phenolic compounds. The antioxidant and antiproliferative effect on leukaemic cell lines with high selectivity indices have been reported already (Larbie et al., 2015). The current study evaluates the hepatoprotective effect of hydroethanolic extract of leaves of F. pumila in CCl4-induced liver damage in rats following

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the observed antioxidant activity and high phenolic content in previous studies. MATERIALS AND METHODS Chemicals The chemicals used in this study include Carbon tetrachloride (CCl4) (Sigma-Aldrich, Germany) and Silymarin (Legalon, Flordis, Australia). Biochemical kits for the determination of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), bilirubin (total and direct), high density lipoprotein (HDL) cholesterol and triglycerides were obtained from Fortress Diagnostics (UK).

conditions of temperature and humidity and had free access to standard feed (AGRICARE, Kumasi-Ghana) and freshly prepared distilled water except an overnight fast before sacrificing. In experimental grouping of the animals, their body weights and sexes were taken into consideration to achieve approximately equal conditions among the groups. The animals were identified by colour codes made on their tails using permanent markers. All animal experiments were conducted in accordance with the guidelines of the Committee for the Purpose of Control and Supervision of Experiment on Animals (CPCSEA, New Delhi, India) and guide for care and use of laboratory animals (Washington, US).

Plant Material and Extract Preparation Evaluation of Hepatoprotective Activity The leaves of Ficus pumila were collected from the Republic Hall on the Kwame Nkrumah University of Science and Technology (KNUST) campus (Kumasi, Ghana) and authenticated (KNUST/HM1/2014/L093) at the Department of Herbal Medicine, KNUST, in the month of November, 2014. The leaves were washed, shade-dried, milled. The 50% hydroethanolic extraction of the plant leaves was carried out by suspending 100 g of the powder of Ficus pumila leaves in 1000 ml of 50% ethanol (50:50, ethanol: water, v/v). The extraction was allowed to stand for 24 hours at room temperature on a shaker (Gallenkamp, UK) and extraction was repeated three times, after which rotary evaporation (Buchi R205, Switzerland) was done. The hydroethanolic extract was freeze-dried (Labconco, England) to obtain the F. pumila hydroethanolic extract (FPE) and transferred into zip locks. It was reconstituted in normal saline for administration. Animals Sprague–Dawley rats of either sex weighing between 170 and 210 g were used for the study. They were obtained from the animal facility of the University of Ghana Medical School, Accra, Ghana. Animals were housed in aluminium cages suitably bedded with wood shaving. They were maintained under standard

Five groups of four animals each were used for the study. Hepatoprotective activity of FPE against CCl4 was assessed by methods described by Arthur et al. (2012). Group 1 served as normal control and received 1 ml/kg normal saline throughout the duration of the experiment. Hepatotoxicity was induced in the other groups with 1 ml/kg CCl4 diluted with olive oil (1:1 vol/vol) administered orally. All the animals of groups II–V were treated with CCl4 for two successive days (2nd to 3rd day by oral route). Group II animals were maintained as CCl4 control without any drug treatment. Groups III and IV were pretreated with 100 and 250 mg/kg FPE, respectively, by oral route. Group V animals were treated with 75 mg/kg silymarin standard. The drug (FPE and silymarin) treatments were administered orally from the 1st to 7th day with concurrent administration of CCl4 on the 2nd and 3rd days. Effect of treatment on body weight Rats in all groups were weighed on the first day (D0) and at the beginning of each day: D1, D2, D3, D4, D5, D6 and D7. The percentage (%) change in body weight was calculated using the formula (Arthur et al., 2011): %

Change

in

Body Weight Ă— 100%

đ??šđ?‘–đ?‘›đ?‘Žđ?‘™ đ?‘Šđ?‘’đ?‘–đ?‘” đ?‘•đ?‘Ąâˆ’đ??źđ?‘›đ?‘–đ?‘Ąđ?‘–đ?‘Žđ?‘™ đ?‘Šđ?‘’đ?‘–đ?‘” đ?‘•đ?‘Ą đ??źđ?‘›đ?‘–đ?‘Ąđ?‘–đ?‘Žđ?‘™ đ?‘Šđ?‘’đ?‘–đ?‘” đ?‘•đ?‘Ą

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=

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expressed as the mean Âą standard error of mean (SEM). Data was assessed by one-way ANOVA followed by Newman–Keuls multiple comparison test. Values for which p<0.05 were considered statistically significant. Percentage protection was calculated with the formula (Arthur et al., 2012): Percent Protection = đ?‘‰đ?‘Žđ?‘™đ?‘˘đ?‘’đ?‘ đ?‘œđ?‘“ đ?‘‡đ?‘œđ?‘Ľđ?‘–đ?‘› đ??śđ?‘œđ?‘›đ?‘Ąđ?‘&#x;đ?‘œđ?‘™ −đ?‘‰đ?‘Žđ?‘™đ?‘˘đ?‘’đ?‘ đ?‘œđ?‘“ đ?‘‡đ?‘’đ?‘ đ?‘Ą đ?‘†đ?‘Žđ?‘šđ?‘?đ?‘™đ?‘’ Ă— đ?‘‰đ?‘Žđ?‘™đ?‘˘đ?‘’đ?‘ đ?‘œđ?‘“ đ?‘‡đ?‘œđ?‘Ľđ?‘–đ?‘› đ??śđ?‘œđ?‘›đ?‘Ąđ?‘&#x;đ?‘œđ?‘™ −đ?‘‰đ?‘Žđ?‘™đ?‘˘đ?‘’đ?‘ đ?‘œđ?‘“ đ?‘ đ?‘œđ?‘&#x;đ?‘šđ?‘Žđ?‘™ đ??śđ?‘œđ?‘›đ?‘Ąđ?‘&#x;đ?‘œđ?‘™

Assessment of hepatoprotective activity All animals were sacrificed on day 8 following an overnight fast. Incisions were quickly made in the sacrificed animal’s cervical region with the aid of a sterile blade and blood samples collected from the heart and dispensed into gel activated tubes for biochemical assays. The plasma obtained from the blood samples was used for liver function tests [AST, ALT, ALP, GGT bilirubin (total and direct)]. Total cholesterol, HDL and triglycerides were determined using the Cobas Integra 400 Clinical Chemistry Analyzer (Roche, USA).

100% RESULTS Effect of Treatment on Percent Body Weight Change

Effect of Treatment on Liver Weight

The effect of the FPE on the body weight of rats is shown in Fig. 1. There was a rise to a peak within the first three days of experimentation in all the groups of rats. Decrease in body weight was observed in rat groups that received CCl4, an indication of toxicity. Increases in body weight were observed from Day 5 till end of the experiment. Best increases were observed in 100 mg/Kg (7.18 Âą 2.304) and 250 mg/Kg (10.26 Âą 2.98) FPE groups, and lowest decreases in CCl4 (1.66 Âą 0.479) and Silymarin (−2.40 Âą 0.835) groups at termination.

Excised liver of the rats were washed in buffered normal saline, blotted dry and weighed to obtain the absolute liver weights. The relative organ weights were calculated with the formula: Relative Organ Weight = đ??´đ?‘?đ?‘ đ?‘œđ?‘™đ?‘˘đ?‘Ąđ?‘’ đ?‘‚đ?‘&#x;đ?‘”đ?‘Žđ?‘› đ?‘Šđ?‘’đ?‘–đ?‘” đ?‘•đ?‘Ą Ă— 100 đ??ľđ?‘œđ?‘‘đ?‘Ś đ?‘Šđ?‘’đ?‘–đ?‘” đ?‘•đ?‘Ą đ?‘Žđ?‘Ą đ?‘†đ?‘Žđ?‘?đ?‘&#x;đ?‘–đ?‘“đ?‘–đ?‘?đ?‘’ Statistical Analysis Data were analyzed using GraphPad Prism 5 for Windows. The experimental results were

Body Weight Changes 15

Normal CCl4

10

100 mg 250 mg Silymarin

5

-5

7 D

6 D

5 D

4 D

3 D

2 D

1 D

0

0 D

Percent Change in Body Weight

Fig. 1: Effect of treatment on Percent Body Weight changes. Each point represents a mean of 4 animals

Days

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Fig. 2: Effect of treatment on absolute and relative liver weight.

Liver Weight 6

Liver Weight

5

***

4

**

Normal CCl4

***

100 mg 250 mg Silymarin

*** ** * *

3 2 1 0 Absolute Weight

% Relative Weight

Each bar represents a mean ± SEM of 4 animals. Statistical difference; *p<0.05, **p<0.01, ***p<0.001 against Normal.

Effect of Treatment on Absolute and Relative Liver Weight

0.146 g) and 100 mg/Kg FPE (4.52 ± 0.104 g) groups.

The effect of Ficus pumila on absolute and relative weights of CCl4 treated rats is shown in Fig. 2. Administration of the hepatotoxin CCl4 caused significant increase in both the absolute and relative liver weights indicating a toxic effect of CCl4 on the liver. The FPE treatment at all doses could not reverse the liver hypertrophy caused by CCl4. Highest significant increase for the absolute weight was observed in the 250 mg/Kg (4.91 ±

Effect of parameters

treatment

on

biochemical

The effect of CCl4 and FPE treatments on indices of liver function are as shown in Table 1. CCl4 caused a 2-fold increase in the levels of ALT, ALP, GGT and TBil and a 1.5-fold increase in AST and DBil levels, an indication of toxicity. Administration of FPE restored these increases to near normal levels.

Table 1: Effect of treatment on the various parameters Treatment

Normal

CCl4

100 mg/Kg of FPE

250 mg/Kg of FPE

Silymarin

228.98 ± 2.64

288.9 ± 6.66*

228.3 ± 12.03+

253.23 ± 17.47

262.78 ± 18.51

Parameters AST (U/L)

+

89.8 ± 11.877 180.25 ± 3.95** 130.40 ± 9.27 131.75 ± 14.37 123.30 ± 15.06 ALT (U/L) ++ 1.60 ± 0.35 3.05 ± 0.10* 2.3 ± 0.48 1.5 ± 0.18 3.43 ± 0.32** GGT (U/L) 163.4 ± 22.07 238.35 ± 25.71** 201.43 ± 3.03 258.90 ± 9.75*** 163.65 ± 14.34++ ALP (U/L) 2.25 ± 0.23 4.68 ± 1.09*** 2.175 ± 0.05++ 2.075 ± 0.19+++ 3.28 ± 0.61 TBil (μmol/L) 1.48 ± 0.16 2.18 ± 0.20 1.58 ± 0.18 1.7 ± 0.15 2.15 ± 0.22 DBil (μmol/L) 0.80 ± 0.30 2.48 ± 1.03* 0.58 ± 0.23+ 0.38 ± 0.06++ 1.13 ± 0.41 IBil (μmol/L) 1.97 ± 0.09 2.37 ± 0.04* 2.18 ± 0.09 2.26 ± 0.11 2.19 ± 0.12 TChol (μmol/L) 0.86 ± 0.08 0.69 ± 0.06 0.75 ± 0.03 0.72 ± 0.06 0.68 ± 0.11 TTrig (μmol/L) 1.55 ± 0.10 1.54 ± 0.07 1.62 ± 0.08 1.8 ± 0.19 1.86 ± 0.08 HDL (μmol/L) *p<0.05, **p<0.01, ***p<0.001 against Normal; +p<0.05, ++p<0.01, +++p<0.001 against CCl4 group TBil-Total bilirubin: DBil-Direct bilirubin: IBil-Indirect bilirubin

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 217–225

Fig 3: Percentage protection based on Liver function indices (ALT, AST, ALP, GGT, TBil, DBil and IBil)

Percentage Protection (%)

90 80 70 60 50

40 30 20 10 0 100 MG

250 MG

SILY

Treatments

Percentage Protection of FPE against CCl4 The various treatments recorded distinct percentage protections based on major indicators of liver function: ALT, AST, ALP, GGT, total, direct and indirect bilirubin (Figure 3). FPE at 100 mg/Kg recorded the best protection (79.87%), followed by 250 mg/Kg (70.46%). Silymarin had the least protection. DISCUSSION The effect of a toxic nature (such as CCl4) is always evident in study subjects by the decrease in body weight due to disturbance in energy metabolism. There was significant increase in the body weight of all the animals from D1 to D3 due to the fact that they received food and water ad libitum. Administration of the CCl4 on D2 and D3 caused sharp decreases in all groups which received the CCl4. This signified the induction of hepatotoxicity. Weight loss is inevitable when animals lose appetite due to disturbance in the carbohydrate, protein or fat metabolism caused by the administration of the toxic material (Klaasen, 2001). Body weight increase was observed from D5 to the end of the experiment in all groups except for the 100 mg/Kg FPE group, which showed body weight increase starting from D6, attributable

to the lower concentration of phytochemicals in the extract required for better pharmacological effect and physical weight gain. At termination, 250 mg/kg had better effect on weight changes (10.26 ± 2.98%). Similar observations were made in previous studies (Arthur et al., 2012). There was significant increase in liver weight upon administration of the CCl4 compared to the normal group due to the toxic action of this hepatotoxin. The toxic effect of CCl4 in all groups receiving it was due to free radical formation which caused lipid peroxidation of cellular membranes (Raju et al., 2003). The liver weight gain is explained by a mechanism leading to the infiltration of the liver by inflammatory cells subsequently resulting in liver necrosis due to the effect of CCl4 intoxication. The 100 and 250 mg/Kg FPE treatment groups recorded significantly higher absolute liver weights compared to the normal and CCl4 groups. The hypertrophy observed could be attributed to the potential of the Ficus pumila extract to enhance the regenerative property of the liver to produce more hepatocytes to mitigate the toxic effect of the CCl4, as observed in the 100 (4.52 ± 0.104) and 250 mg/Kg (4.91 ± 0.146 μmol/L) FPE groups. The lower triglyceride levels observed is an indication that the hypertrophy was not

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 217–225

due to the accumulation of fats but rather accumulation of dead cells, as the lipid peroxidation causing toxicity in the liver was halted. The administration of CCl4 resulted in significantly elevated levels of total cholesterol in the blood serum. The 100 mg/Kg (2.18 ± 0.078 μmol/L), 250 mg/Kg (2.26 ± 0.107 μmol/L) and silymarin (2.19 ± 0.122 μmol/L) had reduced levels compared to the CCl4 (2.37 ± 0.043 μmol/L). Cytochrome P450 acts on the hepatotoxin (CCl4) to produce radicals that bind covalently to the macromolecules, and induce lipid peroxidation of the membrane lipids of the endoplasmic reticulum rich in polyunsaturated fatty acids, causing an increase in cholesterol levels in the CCl4 group. Results of FPE groups indicated a cholesterol lowering potential. High density lipoprotein (HDL) plays a protective role as it reverses cholesterol transport, inhibiting the oxidation of low density lipoprotein (LDL) and neutralizing the atherogenic effects of oxidized LDL (Navab et al., 2001). In the current study, the total triglycerides were found to have decreased while the HDL levels increased with the 250 mg/Kg FPE group (0.72 ± 0.063 μmol/L). The major bioactive components in the extract responsible for the antihyperlipidaemic properties may be directly or indirectly associated with free radical scavenging abilities. This is supported by previous studies which indicated that 50% hydroethanolic extract of FPE is rich in polyphenols and has significant antioxidant activity (Larbie et al., 2015). In the present study, the levels of AST (288.9 ± 6.66 U/L), ALT (180.25 ± 3.95 U/L), ALP (238.35 ± 25.710 U/L) and GGT (3.05 ± 0.10 U/L) were significantly higher (p<0.050.001) in the CCl4 group as compared to the normal group, while the drug treated groups showed a decrease in concentration. The elevated levels of these enzymes indicate liver damage. The hepatoprotective activities of 100 and 250 mg/Kg FPE were monitored by reduced levels in marker enzymes which gave a conclusive idea on the improved functional

state of the liver (Rao and Mishra, 1997) attributable to the antioxidant property of F. pumila (Larbie et al., 2015; Sirisha et al., 2010). In the GGT determination, the CCl4 group recorded elevated levels indicating induced hepatotoxicity (Tate and Meister, 1985) while the 250 mg FPE significantly decreased the GGT levels indicating hepatoprotection. The direct bilirubin in the CCl4 group (4.68 ± 0.202 μmol/L) was higher compared to the normal group (1.48 ± 0.160 μmol/L). This indicates liver malfunctioning as the level of unconjugated or indirect bilirubin in the serum of the group treated with the hepatotoxin CCl4 (2.48 ± 1.031 μmol/L) increased compared to the normal (0.80 ± 0.30 μmol/L). Increase in the concentration of unconjugated (indirect) bilirubin in the blood may result from a defect in the ability of the liver to conjugate bilirubin for excretion (Iniaghe et al., 2008). The 100 mg/Kg (0.58 ± 0.229 μmol/L) and 250 mg/Kg (0.38 ± 0.063 μmol/L) FPE caused significant decreases in unconjugated (indirect) bilirubin levels. The ability of the FPE to improve the conjugating function of the liver suggests that the extracts may activate the constitutive androstane receptor which is a key regulator of bilirubin clearance in the liver (Arthur et al., 2012). In response to elevated bilirubin levels, the constitutive androstane receptor activates expression of multiple components of the bilirubin clearance pathway resulting in increased clearance due to general glycosides (Moore et al., 2004). It could also be attributable to the presence of general glycosides in the crude extract (Larbie et al., 2015) which could be metabolized to glucuronic acid for glucuronidation of bilirubin in humans and rats for excretion (Arthur et al., 2012). The ability of the FPE at 100 and 250 mg/Kg to significantly decrease the concentration of serum total bilirubin when compared to the CCl4 group suggests its potential for bilirubin clearance from the serum, and therefore act as an anti-jaundice agent.

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 7 | July 2016 | 217–225

A comparative analysis of the two doses of FPE and silymarin showed the 100 mg/Kg (79.87%) to have a better protection than the 250 mg/Kg dose (70.46%) compared to standard Silymarin (46.04%). This means that though the FPE showed significant hepatoprotective activity, the dosage at which it is administered is also important.

CONCLUSION This study showed that 50% hydroethanolic extract F. pumila is an effective hepatoprotective agent which brings about functional improvement of liver function. This can be exploited as a therapeutic agent against liver diseases.

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Conflict of Interest: None Declared

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