EVALUATION OF HEPATOPROTECTIVE ACTIVITY OF CARICA PAPAYA AND FICUS BENGALENSIS LATEX ON THIOACETAMIDE INDUCED HEPATOTOXICITY IN RATS.

Yogendra kumar 1 , Girendra Gautam 2 and Pushpesh Kumar Mishra 3 . 1. Research Scholar, Faculty of Pharmacy, Bhagwant University, Sikar Road, Ajmer, Rajasthan, India. 2. Faculty of Pharmacy, Bhagwant University, Sikar Road, Ajmer, Rajasthan, India 3. Department of Chemistry, Faculty of Pharmacy, Naraina Group of Institutions, Kanpur, U.P., India ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History

Carica Papaya Linn plant belongs to family Caricaceae is a tropical tree. It is native to the tropics of an Americas but now widely cultivated in other tropical regions of the world. Plant is available throughout the year [16]. Carica Papaya is fast growing plant, it measure 7-8m with copious latex and trunk about 20 cm in diameter [17].
Aqueous extracts of unripe Carica papaya have been reported antisickling and reversal of sickling properties [20]. Latex, juice and fruit of Carica Papaya have digestive properties and it is reported in used habitual constipation and chronic diarrhea [21]. Carica Papaya Latex reported wound healing properties [22].

Plant materials
Carica Papaya and Ficus Bengalensis were collected from Mathura, Uttar Pradesh. The authentication and identification was done by (Prof.) Dr. D.K. Singh, Department of Botany, KR (PG) College, Mathura, Uttar Pradesh.

Carica Papaya Latex Collection
Latex was collected locally in early morning 7:00 to 8:00 am, as the flow of latex is low during the day. Collection was done by making 1-2 mm deep vertical incisions on the skin of unripe fruit, but mature fruit. The latex was then dried at room temperature till it became crumbly and non-sticky. The dried latex was triturated using a mortar and pestle.. It was stored at 4-8 ºC until use [22].

Ficus Bengalensis Latex Collection
Latex was collected locally in early morning 7:00 to 8:00 am, as the flow of latex is low during the day. Collection was done by making 1-2 mm deep vertical incisions on the skin [22]. Latex was extracted by maceration process (48h) in methyl alcohol after defatted with petroleum ether at (72h) at room temperature. The extracted was dried by rotatory evaporator under reduced pressure [23]. The dried latex was triturated using a mortar and pestle. It was stored at 4-8 0 C until use.

Phytochemical studies
Phytochemical analysis was carried out on Carica Papaya Latex and Ficus Bengalensis Latex for the detection of various phytochemicals by following standard methods described in practical pharmacognosy by Trease and Evans.

Phytochemical Screening
Phytochemicals are chemical compounds that occur naturally in plants. The term is generally used to refer to those chemicals that may affect health, but are not established as essential nutrients. The presence of tannins, alkaloids, flavonoids, general test for glycosides (reducing sugars), anthraquinones, sterols and saponins were tested by simple qualitative methods (Trease and Evans, 1989) [24].

Animals
Albino rats of either sex (Wister strain) weighing 150-200g and female albino mice weighing 20-25g were used in this study. Animals were used from animal house of Sanjay College of Pharmacy, Mathura, U.P.
The animals were acclimatized for ten days under laboratory conditions. They were housed in polypropylene cages and maintained at 27 ºC ± 2 ºC , relative humidity 65 ± 10% under 12 hours light/ dark cycle. The animals were fed with rodent pellet diet (Gold Mohur Lipton India Ltd.) and water ad libitum.
Ethical clearance for performing the experiments on animals was obtained from the Institutional Animal Ethics Committee (IAEC) and registration number 1334/a/10/CPCSEA of Sanjay College of Pharmacy, Mathura, Uttar Pradesh.

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Toxicity study -Determination of acute toxicity LD 50 The acute toxity for Carica Papaya Latex and Ficus Bengalensis latex were determined in female albino mice. The animals were fasted overnight prior to the experiment, fixed dose method of OECD guideline No. 420; (Annexure 2d)of CPCSEA was adopted for this purpose. Group of three mice were taken for each test dose and 1/10 th of LD 50 cut off value of test latex selected as screening dose for Hepatoprotective activity [25]. Group -I serve as a normal control received Saline 1ml/Kg for 10 days. Group -II were administered a single subcutaneous injection of thioacetamide (100mg/Kg. sc) as a 2% w/v solution in distilled water. Group -III, IV and V were treated orally with Caraca Papaya Latex (dose 400mg/Kg.), Ficus Bengalensis Latex (dose 300mg/Kg.) and Silymarin (dose 100mg/Kg.) once daily for 10 days followed by single subcutaneous injection of Thioacetamide. 24 hours after the Toxin administration, the animals of each group were anaesthetized, the blood were collected directly from the heart. The blood samples were allowed to clot for 20-30 minutes. Serum was separated by centrifugation at 37 0 C. Serum was used for estimation of various biochemical parameters such as SGOT, SGPT Bilirubin, ALP and total protein through the auto analyzer for the study of the toxic effect of thioacetamide.

Biochemical Studies
Estimation of SGPT on blood serum was carried out using AGD Clinipak from AGD Biomedicals Pvt. Ltd. Mumbai.
Estimation of Alkaline Phosphate, Total Protein, SGOT on blood serum was carried out using Diagnostic test kit from Beacon Diagnostic Pvt. Ltd., NAVSARI.
Estimation of Bilirubin on blood serum was carried out using diagnostic kit from SIEMENS Ltd., Vadodara, Gujrat.

Histological studies
The Liver were isolated from the animals and washed with normal Saline. The liver was fixed in Formalin diluted to 10% with normal Saline then processed further for histological studies. The results were analyzed by student t-test.

Statistical analysis
All the values are expressed as mean ±S.D. result were analyzed statistically by analysis of variance (ANOVA) followed by student T-Test was used for determining significance.

Result And Discussion:-
The result of present study reveals that level of parameters like SGPT, SGOT, Bilirubin, and ALP were found to be elevated and decrease total protein in Thioacetamide treated group (dose 100 mg/kg b.w.) whereas treatment along with Carica Papaya Latex (dose 400mg/kg b.w.) and Thioacetamide (dose 100 mg/kg b.w.) group shows significant reduction in above said parameter SGPT (180.7±14.  Histological studies showed that Thioacetamide produced extensive vascular degenerative changes and centrilobular necrosis and Hepatocytes. It was shown in figure no.1 to 5.

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Administration of Thioacetamide treated group at the dose of 100 -300 mg/kg by s.c. route results in the Hepatic damage in animals. Thioacetamide is a Hepatotoxic that is metabolized by CYP 450 enzymes present in liver microsomes and is converted to a toxic reactive intermediate called Thioacetamide (s-oxide) due to oxidation process. Thioacetamide s -oxide induced oxidative stress in the Hepatic cells. It interferes with the movement of RNA from the nucleus to the cytoplasm which may cause membrane injury. It reduce the number of viable Hepatocytes as well as rate of oxygen consumption and also decrease the volume of bile and its content. In the present study groups treated with Thioacetamide100 mg/kg [26].
We found that the treatment with both latex prevented elevation of serum parameter like SGPT, SGOT ,Bilirubin , ALT,and decreased total protein due to Thioacetamide challenge models.
Treatment with both latex found that it improved the liver anatomy / hepatic anatomy with normalization of histopathological observation observe after Thioacetamide induced hepatotoxicity.

Conclusion:-
It is thus concluded that both plant Carica Papaya Latex and Ficus Bengalenesis Latex posseses a hepatoprotective activity against thioacetamide induced hepatotoxicity in rats as evidenced by reduction in serum parameter like SGPT, SGOT, Bilirubin, ALT,and elevation of total protein etc. Whereas Carica Papaya latex was found to be more effective compared to Ficus Bengalenesis Latex.
A finding from the hepatoprotective study indicates that both latex were showed significantly reduced the elevated serum parameter and enhanced the decreased level of total protein induce by hepatotoxin in rats .