Hepatoprotective Activity of Vitex trifolia against Carbon Tetrachloride-induced Hepatic Damage

antipyretic , antibacterial 5 , against and to pharmacological screening for its hepatoprotective reports the hepatoprotective activity of Vitex trifolia against CCl 4 hepatic Aqueous was investigated for hepatoprotective activity against carbon tetrachloride induced liver damage. To assess the hepatoprotective activity of the extracts, various biochemical parameters viz., total bilirubin, total protein, alanine transaminase, aspartate transaminase and alkaline phosphatase activities were determined. Results of the serum biochemical estimations revealed signiﬁ cant reduction in total bilirubin and serum marker enzymes and increase in total protein in the animals treated with ethanol and aqueous extracts. However signiﬁ cant rise in these serum enzymes and decrease in total protein level was noticed in CCl4 treated group indicating the hepatic damage. The hepatoprotective activity is also supported by histological studies of liver tissue. Histology of the liver tissue treated with ethanol and aqueous extracts showed normal hepatic architecture with few fatty lobules. Hence the present study revealed that Vitex trifolia could afford signiﬁ cant protection against CCl 4 induced hepatocellular injury. Viswanathan, et al. : Synthesis and anticancer activity of L-glutamic amides Four N-(benzenesulfonyl)-L-glutamic acid bis(p-substituted phenylhydrazides) were synthesized and evaluated for anticancer activity in vitro in DU-145 and PC-3 prostate cancer and in COLO-205 colon cancer cell lines by MTT assay. The analog with the nitro group substitution exhibited potent activity (% Inhibition 84.7 and 72.0 in DU-145 and PC-3 respectively at 80 µ g/ml concentration). Another series of substituted 1-(benzenesulfonyl)-5-oxopyrrolidine 2-carboxamides (11a-f) were synthesized and evaluated for anticancer activity in vitro in colon (COLO-205), breast (Zr-75-1) and prostate (PC-3) cancer cell lines by MTT assay using adriamycin as standard. Test compounds 11a-c showed potent activity (% Inhibition 61.2 to 79.2 at 20 µ g/ml and 67.2 to 87.2 at 40 µ g/ ml) in PC-3 cell line which is superior to the activity of Adriamycin. In comparison compounds 11d-f were less potent. In Zr-75-1 cell line 11a-e showed % inhibition ranging from 32.4 to 54.9 at 10 µ g/ml concentration while in COLO-205 cell line 11a-f showed poor activity.

and increasing the permeability to ATP through cytoplasmic membrane. Similarly, Rath et al 9 , also reported the anti staphylococcal activity of Juniper and Lime essential oils against methicillin resistant Staphylococcus aureus (MRSA) through inhibition of cell membrane synthesis that corroborates with the findings observed in this investigation. The antibacterial activity of essential oils through membrane inhibition could be attributable to the hydrophobicity of essential oils, enables them to make partitions in the membrane, rendering permeability and leading to leakage of cell contents resulting in death of microbial cells 10-12. In conclusion, this investigation amply proved the antibacterial activity and mechanism of action of Jasminum sambac natural oil and its synthetic blends against E. coli MTCC-443 strain. Leaves were shade dried for a week, powdered Aqueous and ethanol extracts of leaf of Vitex trifolia was investigated for hepatoprotective activity against carbon tetrachloride induced liver damage. To assess the hepatoprotective activity of the extracts, various biochemical parameters viz., total bilirubin, total protein, alanine transaminase, aspartate transaminase and alkaline phosphatase activities were determined. Results of the serum biochemical estimations revealed signifi cant reduction in total bilirubin and serum marker enzymes and increase in total protein in the animals treated with ethanol and aqueous extracts. However signifi cant rise in these serum enzymes and decrease in total protein level was noticed in CCl4 treated group indicating the hepatic damage. The hepatoprotective activity is also supported by histological studies of liver tissue. Histology of the liver tissue treated with ethanol and aqueous extracts showed normal hepatic architecture with few fatty lobules. Hence the present study revealed that Vitex trifolia could afford signifi cant protection against CCl 4 induced hepatocellular injury.
Key words: Vitex trifolia, aqueous and ethanol leaf extracts, CCl 4 , hepatoprotective activity  lobules (fi g. 4). However signifi cant liver protection was observed in the liver sections of ethanol extract treated animals as evident by the presence of normal hepatic cords, absence of necrosis with few fatty lobules (fi g. 5).
CCl 4 -induced hepatic injury is the common model used for hepatoprotective drug screening. The extent of hepatic damage is assessed by the elevated level of biochemical parameters which is attributed to the generation of trichloromethyl free radical which in turn causes peroxidation of lipids of cellular membrane 15 . In the present investigation, preliminary phytochemical analysis of leaf extracts revealed the presence of flavonoids, tannins, saponins, glycosides, steroids and triterpenoids. Flavonoids 16 and triterpenoids 17 are well known for their antioxidant and hepatoprotective activities. In this study ethanol extract showed protective effect against toxicity induced by CCl 4 , which may be attributed to the individual or combined effect of antioxidant and hepatoprotective activity of phytoconstituents present in it. Based on the above results of the pharmacological screening, it can be concluded that the ethanol and aqueous leaf extracts of Vitex trifolia possesses significant hepatoprotective activity, which provides scientific evidence to the ethnomedicinal value of this rare plant genetic resource used by the tribal group of Western Ghats in treating jaundice.
bleeding into sterilized dry centrifuge tubes and allowed to coagulate for 30 min at 37°. The clear serum was separated at 3000 rpm for 10 min and was subjected to biochemical investigation viz., total bilirubin 10 , total protein 11 , serum alanine transaminase, aspartate transaminase 12 and alkaline phosphatase 13 . Results of biochemical estimations were reported as mean±SE of six animals in each group. The data was subjected to one way ANOVA followed by Tukey's multiple comparison tests. P≤0.01 was considered as statistically signifi cant.
The liver samples were excised from the experimental animals of each group and washed with the normal saline. Initially the materials were fixed in 10% buffered neutral formalin for 48 h and processed for paraffin embedding. The sections were taken at 5 µ thickness using microtome, processed in alcohol-xylene series and were stained with alum-haemotoxylin and eosin 14 . The sections were examined microscopically for the evaluation of histological changes.
Effect of ethanol and aqueous leaf extracts of Vitex trifolia on CCl 4 -induced liver damage in rats with reference to biochemical changes in serum is shown in the Table 1. At the end of 14 d treatment, blood samples of CCl 4 -treated animals showed significant increase in the levels of total bilirubin, alanine transaminase, aspartate transaminase and alkaline phosphatase compared to normal control groups but the total protein level decreased refl ecting the liver injury caused by CCl 4 . Whereas blood samples from the animals treated with ethanol and aqueous leaf extracts of Vitex trifolia showed signifi cant decrease in the levels of serum markers and signifi cant increase in total protein to the near normal which are comparable to the values registered in the standard drug treated group of animals, indicating the protection of hepatic cells. Among the two extracts ethanol leaf extract showed significant protection against CCl 4 induced hepatic damage.
Histological profi le of control animal showed normal hepatocytes (fi g. 1). The section of liver of the group II animals exhibited severe intense centrilobular necrosis, vacuolization and macro vesicular fatty changes (fi g. 2). The liver sections of silymarin-treated animals showed normal hepatic architecture (fi g. 3). The liver sections of the animals treated with aqueous extract exhibited moderate accumulation of fatty