Abstract: The hepatoprotective effects of grape seed oil (GSO) against carbon tetrachloride (CCl₄) induced acute liver injury in mice was investigated. The model of acute liver injury was induced by intraperitoneal injection of CCl₄. The body weight and food and water intake were measured, the relevant indexes of serum and liver tissue were detected, and the pathological changes of liver tissues in mice were observed. The results showed that compared mice-treated with CCl₄, the mice were oral administered middle dose (10 mL/kg·d) of GSO (MGSO) and high dose (20 mL/kg·d) of GSO (HGSO) could significantly reduce the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and the levels of total cholesterol (TC), triglyceride (TG) (P<0.05) in serum, could significantly increase the activities of glutathione (GSH), superoxide dismutase (SOD) and reduce concentrations of malondialdehyde (MDA) (P<0.05), and could significantly improve the pathological changes of liver tissue. Therefore, GSO had significant hepatoprotective effects against CCl₄ induced acute liver injury in mice, and the hepatoprotective effects was stronger with the increasing dose．