This study aimed at investigating the impact of Diazepam on serum liver enzyme, oxidative stressm, and som antioxidant markers in male rabbits. Methods: A total of 20 growing rabbits (7.5 weeks old) reared under high ambient temperature were divided into two equal groups, 10 rabbits each. The first group control administered with normal saline and the second group Diazepam-treated rats (50 mg/kg b. w. orally) for 30. Blood samples were withdrawn to measure serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), Glutathione (GSH), malondialdehyde (MDA) levels, and catalase (CAT), activities were assayed. Results: In Diazepam -treated group, significant increases in aspartate aminotransferases, and alanin aminotransferases activities, malondialdehyde (MDA) levels and significant decreases in catalase (CAT), glutathione activities levels were determined compared to the control group. Conclusion: Diazepam administration produces noticeable biochemical changes in a dosedependent manner associated with increased liver enzyme and oxidative stress markers and decreased antioxidative activity.