Background: Medicinal herbs are significantly effective against a variety of liver disorders and Trapa natans was traditionally used for the treatment of anti-inflammatory, pain disorder, and various types of hepatic ailment. Objective: The purpose of this study was to evaluate the hepatoprotective activity of T. natans fruit peel extract against antitubercular drugs (isoniazid + rifampicin [INH + RIF])-induced hepatotoxicity in rats. Materials and Methods: Liver toxicity was induced by INH + RIF at a dose level of 50 mg/kg each, intraperitoneally. for 15 days. Fifty percent ethanolic extract of T. natans (TNE) at a dose of 200 and 400 mg/kg was administered orally once daily for 15 days. The hepatoprotective activity was assessed using various biochemical parameters such as aspartate transaminase, alanine transaminase, alkaline phosphate, lactate dehydrogenase, albumin, cholesterol, and bilirubin. Furthermore, in vivo antioxidant activities and histopathological investigation were performed to assess hepatoprotective activity. Results: Obtained results demonstrated that the level of liver marker enzymes and antioxidant parameters were significantly altered by INH + RIF treatment. Treatment with T. natans peel extract causes significant (P < 0.01 to P < 0.001) reduction in liver injury & normalized all altered liver marker enzymes. In addition, TNE significantly normalized the activity of antioxidant enzymes, namely, lipid peroxidation (P < 0.01 to P < 0.001), reduced glutathione (P < 0.05 to P < 0.001), superoxide dismutase (P < 0.05 to P < 0.001), and catalase (P < 0.01 to P < 0.001) in the liver tissue of INH + RIF-treated groups. Histological observations of the liver tissues correlated with the biochemical observations. Conclusion: These findings powerfully support that the protective effect of T. natans fruit peel extract against liver injury which may be attributed to its hepatoprotective activity due to normalizes the altered liver marker enzymes and antioxidant defense status and thereby contributed to its antihepatotoxic potential.