Objective: The aim of this study was to evaluate the hepatoprotective and antioxidant effects of Withania somnifera against Paracetamol-induced liver injury in rats. Methods: In the present study, the protective effect of Withania somnifera was investigated against Paracetamol-induced hepatotoxicity and compared with Silymarin, a standard hepatoprotective reference drug. The rats received a single dose of paracetamol (900 mg/kg body weight, i.p.); Withania somnifera (500 mg/kg body weight and 1000mg/kg body weight, p.o.) and Silymarin (25 mg/kg body weight, p.o.) were administered 30 min after the injection of paracetamol. Liver marker enzymes (Aspartate Transaminase, Alanine Transaminase and Alkaline Phosphatase), Total Protein content, Bilirubin, Antioxidant status (Reduced Glutathione, Superoxide Dismutase, Catalase and Glutathione-S-Transferase) were evaluated and histopathological analysis was done for the control and experimental rats. Results: Paracetamol treatment leads to elevated levels of liver marker enzymes and bilirubin and there was deterioration in total protein content, histological observations and antioxidant status. However, treatment with Withania somnifera significantly reversed (p < 0.05) the above changes compared to the control group as observed in the paracetamol-challenged rats. Conclusion: The results clearly demonstrate that Withania somnifera possesses promising hepatoprotective effects through its antioxidant activity and hence suggests its use as a potential therapeutic agent for protection from paracetamol overdose.