Hepatoprotective Activity of Methanolic Extract of Whole Plant of Rhynchosia beddomei in Wistar Rats

In the present study, the methanolic extract of Rhynchosia beddomei leaves was evaluated for its hepatoprotective effect against Ranitidine and Paracetamol induced hepatic injury in rats. Alteration in the levels of biochemical markers of hepatic damage like SGOT, SGPT, ALP and total proteins were evaluated. Paracetamol (500 mg/kg) and Ranitidine (150 mg/kg) induces hepatotoxicity in 7 and 21 days respectively and enhances the SGPT, SGOT, ALP, liver weight and reduced total proteins. Treatment with methanolic extract of Rhynchosia beddomei (150 mg/kg and 300 mg/kg) has brought back the altered levels of biochemical markers significantly to the near normal levels in the dose dependent manner. The results were supported by histopathological studies of liver tissue. Phytochemical analysis of Rhynchosia beddomei indicated the presence of alkaloids, phenolics, saponins, flavonoids and polysaccharides and its hepatoprotective potential may be due to the presence of flavonoids. Keyword: Rhynchosia beddomei; Biochemical markers; SGOT; SGPT; ALP Suvarchala Reddy et al 379 J Pharm Chem Biol Sci , September-November 2015; 3(3): 378-387 necrosis, increase in tissue lipid peroxidation and depletion in the tissue GSH levels. In addition serum levels of many biochemical markers like SGOT, SGPT, ALP and bilirubin are elevated [2]. Herbs play a major role in the management of various liver disorders along with other system associated diseases. Hepatotoxicity is very common aliment resulting in serious debilities ranging from severe metabolic disorders to even mortality [3]. Plant derived natural products such as flavonoids, terpenoids and steroids have received considerable attention in recent years due to their diverse pharmacological properties including antioxidant and hepatoprotective activity [4,5]. Herbal medicines are known to play an important role in the treatment of various elements including liver disorders and many traditional practioners have claimed that numerous medicinal plants can be extensively used for the alleviation of different types of liver disorders [6]. Inspite of phenomenal growth of modern medicine there are no synthetic drugs are available for the treatment of hepatic disorders. However there are several herbs/herbal formulation claimed have possess beneficial activity in treating hepatic disorders. Rhynchosia beddomei Baker, is plant of natural origin belongs to the family Fabaceae is a rare and endemic plant restricted to Seshachalam (Gogarbham hills) of Eastern Ghats of Andhra Pradesh, India. Different parts of R. beddomei plant have been found to possess medicinal properties; it possesses wound healing [7], in diuretic [8], anti-inflammatory, carcinoma, antioxidant [9] and antidiabetic activity [10]. R. beddomei contains the volatile oils was first reported. The phytochemical properties of the leaves and reported the presence of alkaloids, indole alkaloids, anthracene glycosides, anthraquinones, carotenoids, coumarins, dihydrochalcones, fatty acids, flavonoids, flavones, flavonols, steroids and triterpenoids [11]. So in the present investigation whole plant of R. beddomei was selected and screened for hepatoprotective activity. MATERIALS AND METHODS Plant material and preparation of extract R. beddomei whole plant was collected from Seshachalam hills and authenticated by Dr. K. Madhava Chetty, Assistant Professor in Department of Botany, Sri Venkateshwara University, Tirupati, Chittoor district, Andhra Pradesh. The crude plant material was dried under shade and powdered mechanically to coarse powder. The coarsely powdered plant material (500 gm) was subjected to extraction with methanol using simple distillation. The extract was evaporated to semisolid mass and subjected to preliminary phytochemical investigation. Animals Studies Healthy adult Wistar Albino rats of 180-250 gm were selected for the study. The animals were obtained from Gentox laboratories, Hyderabad. The animals were housed according to CPCSEA guidelines (under standard temperature condition). They were given a pellet diet and water ad libitum. The ethical clearance was obtained from Institutional Animal Ethics Committee (IAEC) before the experiment (Reg. no.1175/ac/08/CPCSEA). Acute toxicity studies (OECD 423) An acute toxicity study was performed on methanol extract following OECD guidelines (423). The dosage for the pharmacological studies was selected as 1/10 of the highest dose (3000 mg/kg) administered. Experimental design Ranitidine induced hepatotoxicity In this study Male Wistar Albino rats of 150 – 200 gm are taken. Total of 30 animals are involved. Animals are divided in to six groups as Group I as (Control) which receives normal distilled water, Group II as (Negative control) Suvarchala Reddy et al 380 J Pharm Chem Biol Sci , September-November 2015; 3(3): 378-387 which receives Ranitidine at a dose of 150 mg/kg bd.wt for 21 days through i.p route, Group III as (Test I) which receives Ranitidine for 21 days and MERB at a dose of 150 mg/kg bd.wt for 10 days through oral route, Group IV as (Test II) which receives Ranitidine for 21 days and along with it MERB at a dose of 300 mg/kg for 10 days through oral route, and Group V as (Standard) which receives Ranitidine for 21 days and the standard drug Liv-52 at a dose of 5 ml/kg bd.wt through oral route [12,13]. On the 11th day, 18 h after the dose of drugs, all the animals were anaesthetized under light ether anesthesia and the blood was collected from retro orbital sinus using a heparinized capillary tube for the study of biochemical parameters. Paracetamol induced Hepatotoxicity In this study Male Wistar Albino rats of 150 – 200 gm are taken. Total of 30 animals are involved. Animals are divided in to six groups as Group I as (Control) which receives normal distilled water, Group II as (Negative control) which receives Paracetamol at a dose of 500 mg/kg bd.wt for 7 days through i.p route, Group III as (Test I) which receives Paracetamol for 7 days [6,14] and MERB at a dose of 150 mg/kg bd.wt for 7 days through oral route, Group IV as (Test II) which receives Paracetamol for 7 days and along with it MERB at a dose of 300 mg/kg for 7 days through oral route, and Group V as (Standard) which receives Paracetamol for 7 days and the standard drug Liv-52 at a dose of 5 ml/kg bd.wt through oral route. On the 8th day, 18 h after the dose of drugs, all the animals were anaesthetized under light ether anesthesia and the blood was collected from retro orbital sinus using a heparinized capillary tube for the study of biochemical parameters. In vivo Antioxidant studies Antioxidant studies such as superoxide scavenging activity, Lipid peroxidation (LPO) assay, GSH (glutathione) assay and Catalase assay were evaluated. [6] Histopathological studies A portion of liver tissue (liver slices) of normal control, ranitidine control, paracetamol control and treated groups of rats with Liv-52 and MERB (150 mg/kg) and MERB (300 mg/kg) were stored in containers for 12 hours in 10% formalin solution and subjected to histopathological studies. Observed microscopically for histopathological changes i.e., normal liver, damaged and recovered liver was studied and compared. [15] The results were shown in Figure 1 & Figure 2. Statistical analysis All the values were expressed as mean ±SEM. The data’s were statistically analyzed by one way ANOVA followed by Dunnett’s t-test and values p< 0.05 was considered to be significant.