The present study determined the possible protective effect of pumpkin seed oil (PSO) on histopathological changes in liver tissue and serum alanine aminotransferase (ALT) activity, glucose, Non-Esterified Fatty Acids (NEFA) and triglyceride (TG) levels on chronic liver injury induced by carbon tetrachloride (CCl4) in rats. A total of 80 Wistar rats were divided into eight groups of ten rats each: the first group served as the control, group II was given intraperitoneal 0.2 mL/kg CCl4, twice a week for 8 weeks. Groups III-V were treated daily with PSO for 8 weeks (1, 2 and 3 mL/kg respectively). Groups (VI-VIII) were administered with intraperitoneal CCl4 (0.2 mL/kg) twice a week and simultaneously PSO for 8 weeks (1, 2 and 3 mL/kg, respectively). Groups VII and VIII showed a partial decrease of steatosis in the hepatocytes while the findings in the Group VI were similar to Group II. Serum glucose, TG and NEFA levels, which were increased as a result of liver toxicity, decreased significantly with the treatment of 1, 2 and 3 mL/kg PSO. In addition, serum ALT activity also decreased in CCl4 groups given 2 and 3 mL / kg PSO.As a result, using of PSO at a dose of 3 mL/kg is more effective than 1 and 2 mL/kg on the some biochemical and histopathological changes in CCl4 induced liver damage. Further investigations on dosage and duration that produce the best result without side effects, need to be performed.