Abstract: The present study was intended to evaluate the hepatoprotective activity on ethanolic extract of leaves of Vitex negundo was determined by using Wistar Albino rats in male sex. The V. negundo Linn. is a natural plant product, in it’s the leaves are used with the added advantage to revert Ibuprofen induced hepatotoxicity. Oral administration of ethanol extract of V. negundo (100 and 300 mg) produced a significant and dose dependent inhibition to the acute hepatotoxic induced rats and various parameters were analyzed, when compared with negative control V. negundo showed that the significant activity in 300 mg/kg/b.wt. They exhibited a significant inhibition of hepatic toxicity by using various marker enzymes and the histopathological analysis. The inhibitory effect of the V. negundo on hepatotoxicity was compared to that of positive control group. The various parameters such glucose, protein, triglycerides, bilirubin, urea, creatinine, ALP, ACP, SGPT, SGOT and histopathological parameters was measured by dissection the rats. A significant index and values were observed in the acute assays; an effective significant alteration in all biochemical and histopathological sections was observed. From these results, concluded that the V. negundo having the potential effectiveness at the dose of 300 mg/kg/b.wt., (p<0.01) significance in a dose dependent manner. These results suggest that leaves of V. negundo having the hepatoprotective activity, which support the hepatic cells protection.