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Hepatoprotective Activity of Adina Cordifolia Against Ethanol Induce Hepatotoxicity in Rats

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Hepatoprotective Activity of Adina Cordifolia Against Ethanol Induce Hepatotoxicity in Rats Sharma et al., International Current Pharmaceutical Journal 2012, 1(9): 279-284 International Current http://www.icpjonline.com/documents/Vol1Issue9/09.pdf Pharmaceutical Journal ORIGINAL RESEARCH ARTICLE OPEN ACCESS Hepatoprotective activity of Adina cordifolia against ethanol induce hepatotoxicity in rats *A Sharma1, B Sangameswaran2, V Jain3, M S Saluja1 1Research Scholars, Department of Pharmacy, Suresh Gyan Vihar University, Jaipur, Rajasthan, India 2Principal, SSM College of Pharmacy, Jambai Village, Bhavani Taluk, Erode-638 312, Tamil Nadu, India 3TIT College of Pharmacy, Bhopal, India ABSTRACT The acetone (AEAC) and aqueous extracts (AQEAC) of Adina cordifolia, belonging to the family Rubiaceae, were studied for hepatoprotective activity against Wister rats with liver damage induced by ethanol. It was found that AEAC and AQEAC, at a dose of 500 mg/kg body weight exhibited hepatoprotective effect by lowering the Serum Glutamate Pyruvate Transaminase (SGPT), Serum Glutamate Oxaloacetate Transaminase (SGOT), alkaline phosphate and total bilirubin to a significant extent and also significantly increased the levels of total protein. The hepatoprotec- tive activity was also supported by histopathological studies of liver tissue. Since results of biochemical studies of blood samples of ethanol treated rats showed significant increase in the levels of serum enzyme activities, reflecting the liver injury caused by ethanol and blood samples from the animals treated with AEAC and AQEAC showed significant decrease in the levels of serum markers, indicating the protection of hepatic cells against ethanol induced hepatocellular injury. The effects of AEAC and AQEAC were comparable with standard drug silymarin. Key Words: Silymarin, SGOT, SGPT, alkaline phosphate, total bilirubin, total protein. INTRODUCTIONINTRODUCTION (Sharma et al., 1991; Subranonium & Pushpangadan, Liver is one of the largest organs in human body 1999). Thus liver diseases are some of the fatal and the chief site for intense metabolism and disease in the world today. They pose a serious excretion. So it has a surprising role in the mainten- challenge to international public health. Modern ance, performance and regulating homeostasis of medicines have little to offer for alleviation of the body. It is involved with almost all the biochem- hepatic diseases and it is chiefly the plant based ical pathways to growth, fight against disease, preparations which are employed for their treat- nutrient supply, energy provision and reproduction ment of liver disorders. But there are not much drug (Ward and Daly, 1999). The major functions of the available for the treatment of liver disorders (Karan liver are carbohydrate, protein and fat metabolism, et al., 1999; Chaterrjee, 2000). Therefore, many folk detoxification, secretion of bile and storage of remedies from plant origin are tested for its poten- vitamin. Thus, to maintain a healthy liver is a crucial tial antioxidant and hepatoprotective liver damage factor for overall health and well being. But it is in experimental animal model (Rubinstein, 1962; continuously and variedly exposed to environmen- Suja et al., 2002). The medicinal use of many plants tal toxins, and abused by poor drug habits, and (as hepatoprotectives) like Andrographis paniculata, alcohol and prescribed & over-the-counter drug Azadirachta indica, Cassia fistula, Elephantopus scaber, which can eventually lead to various liver ailment Hibiscus rosasinensis, Phyllanthus debilis, Picrorrhiza like hepatitis, cirrhosis and alcoholic liver disease kurroa has been reported in the literature (Rajesh et al., 2001; Anandan et al., 1999). *Corresponding Author: Ajay Sharma, Assistant professor Department of Pharmacy Adina cordifolia, (Rubiaceae) (commonly known as Suresh Gyan Vihar University Haridru) is found throughout central and south India Jagatpura, Jaipur, Rajasthan, India. to Srilanka. The 7-hydroxycoumarin-1 and 7-β-D- E-mail: [email protected] glucosylcoumarin-2 were isolated from the root bark Contact No.: +91- 09827687111 © 2012 Sharma et al.; licensee Saki Publishing Club. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nd/3.0/), which permits unrestricted use (including commercial use), distribution and reproduction of the work in any medium, provided the original work is properly cited and remain unaltered. of Adina cordifolia, having antiamoebic activity Table 1: Evaluation of hepatoprotective activity by (Firdoos et al., 2009). According to ayurveda, the bark ethanol induced hepatotoxicity in rats. of this plant is used in liver disorders (Yoganarasim- Group Treatment han 1996). Adina cordifolia was also reported for its antiulcer potential activity (Kasinadhuni et al., 1999), I Received water (5ml/kg. p.o) for 21 days once daily, and served as normal control. antifertility activity (Sabir et al., 1970), anti- inflamma- I & II Received water (5 ml/kg. p.o) for 21 days tory and anti-nociptive activity (Jain et al., 2006). once daily and 40% ethanol (v/v, 2.0ml/l00g body wt, p.o.) for 21 days. I, II & III Received standard drug silymarin (25 METHODMATERIALS AND AND MATERIALS METHODS mg/kg. p.o.) for 21 days once daily and Procurement and Authentification of the Plant 40% ethanol (v/v, 2.0ml/l00g body wt, p.o.) The leaves of Adina cordifolia were collected from for 21 days. local area of village Ingoriya, Ujjain, Madhya I, II, III & IV Received AEAC (500 mg/kg) 21 days once Pradesh and it was authenticated by Dr. S. K. daily and 40% ethanol (v/v, 2.0 ml/l00 g body wt, p.o.) for 21 days. Billore, at Vikram University, Ujjain, Madhya V Received AQEAC (500 mg/kg) 21 days Pradesh. The voucher specimen (MIPS/N/012/2010) once daily and 40% ethanol (v/v, was deposited at Department of Pharmacognosy, 2.0ml/l00g body wt, p.o.) for 21 days. Mahakal Institute of Pharmaceutical Studies, Ujjain, Rats were divided into 5 groups of 6 animals (n=6) in each Madhya Pradesh for future reference. (Kapoor et al., 1994) Preparation of extracts of Adina cordifolia The powdered leaves (500g) were extracted by using SGPT and ALP etc. were obtained from Span acetone by sequentially extracted using petroleum Diagnostics Ltd., India. ether, chloroform, acetone and ethanol in Soxhlet apparatus. Whereas aqueous extract was obtained Acute toxicity studies by cold maceration processes. After about forty Healthy Wistar albino rats of either sex weighing siphons of each solvent extraction step, the materials 100-150 g maintained under standard laboratory were concentrated by evaporation (Farnsworth, conditions were used for acute oral toxicity test 1966). according to Organization for Economic Co- operation and Development guidelines 423. Ani- Animals mals were observed individually at least once Wistar albino rats (150-200g) used in studies, was during first 30 min after dosing, periodically during procured from Central Drug Research Institute, first 24 h (with special attention during the first 4 h) Lucknow, India. The animals were fed with stan- and daily thereafter for period of 3 days (OECD, dard pellet diet (Hindustan lever Ltd. Bangalore) 1996). Observations were done daily for changes in and water ad libitum. All the animals were acclima- skin and fur, eyes, mucus membrane (nasal), tized for a week before use. The experimental respiratory rate, circulatory signs (heart rate), protocols were approved by Institutional Animal autonomic effect (salivation, lacrimation, perspira- ethics Committee after scrutinization. Animals were tion, urinary incontinence and defecation) and received the drug by oral gavage tube. All the central nervous system (drowsiness, gait, tremors animals were care of under ethical consideration as and convulsion) changes. per the CPCSEA guidelines (CPCSEA, 2003) with regular inspections of rats. The laboratory condi- Assessment of hepatoprotective activity tions duly undertaken by registered veterinary After 24 h of ethanol administration, on 22nd day, practitioner. blood was obtained from animals by puncturing retro orbital plexus. Blood samples were allowed to clot for Chemicals 45 min at room temperature. Serum was separated by All the chemicals and solvents were of analytical centrifugation at 2500 rpm at 30°C for 15 min and grade. Silymarin was obtained as gift sample from utilized for the estimation of various biochemical Micro Lbs, Goa, India. Standard kits for SGOT, parameters including SGOT & SGPT (Reitman et al., 280 Table 2: Effect of AEAC and AQEAC on ethanol induced hepatotoxicity in rats. Treatment/ Dose SGPT(µ/min/l) SGOT(µ/min/l) ALP(µ/min/l) Total Bilirubin mg/dl Total Protein gm/dl Normal 62.0±3.71 168.04±2.80 190.0±8.01 0.38 ±0.06 9.57±0.24 Induced(ethanol) 98.75±8.86* 258.42 ±4.24* 244.76±8.82* 6.42±8.66* 5.40±8.46* Standard (silymarin 63.76 ±4.63** 176.28±8.47** 194.27±4.27** 0.45±2.82** 9.81±4.26** 25mg/kg) AEAC (500mg/kg) 79.88±8.22** 196.28± 4.24** 210.46±8.22** 0.68 ±4.62** 8.28± 8.12** AQEAC (500mg/kg) 68.28 ±6.76*** 186.84± 4.26*** 198±8.44*** 0.56.0±2.20*** 6.18±7.48*** Values are mean ±SEM, n= 6. (One way ANOVA Followed by Dunnette multiple Comparisons test) Statistically significance of ** P<0.01, *** P<0.001, when compared with respective control 1957), ALP (Kind et al., 1954), serum bilirubin (Amour mg/kg. This finding suggests that the AEAC and et al., 1965) and serum protein (Lowry et al., 1951) After AQEAC were safe or non-toxic to rats and hence collection of blood samples, the animals were sacri- doses of 500 mg/kg, p.o. were selected for the study. ficed under deep ether anesthesia. Effect of AEAC and AQEAC on serum marker Morphological parameters like weight of animals, enzyme levels weight of liver have also been used to evaluate the There was a significant elevation in the levels of protective effect of the drug. Hepatoprotective serum marker enzymes like SGOT, SGPT etc, chemical causes loss in liver weight/100 gm body content of ethanol intoxicated animals.
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