To evaluate effect of Polyherbal Formulation (PHF) on drug induced hepatotoxicity in rats and assessment of any possibility of co administration of PHF along with such hepatotoxic drug. Hepatotoxicity in rat was induced by Methotrexate (1mg/kg p.o./week for 3 weeks) and protective effect of PHF (0.25ml/kg/p.o. and 0.5ml/kg/p.o. either along with drug or followed by inducing hepatotoxicity) was measured by estimating marker enzymes for liver function like Aspartate aminotransferase, Alanine aminotransferase, Alkaline phosphatase and . glutamic transpeptidase. Oxidative stress markers like lipid peroxidation, reduced glutathione, super oxide dismutase and catalase. Protein profile likes Total bilirubin, direct bilirubin, Total albumin and Total protein. Histopathological study was carried out to confirm hepatotoxicity. Methotrexate induced hepatotoxicity characterized by significant (P<0.001) increase in marker enzymes for liver function and oxidative stress. Protein profile significantly depleted. Administration of PHF either along with Methotrexate or followed by inducing hepatotoxicity significantly improved (P<0.001) the level of marker enzymes for liver function, oxidative stress and depleted proteins profile. The study suggests protective role of PHF in drug induced hepatotoxicity and it can be utilized to treat the hepatotoxicity with long-term clinically useful drugs.