Drug induced liver injury has been regarded as the main cause of acute liver injury in many countries. Objectives: To evaluate the protective action of selenium against hepatotoxicity caused by methotrexate in mice. Study Design: Randomized, single blinded study. Methodology: Present study was done on 18 mice (n = 18) after taking approval from Ethical Review Committee. Administration of methotrexate induced hepatotoxicity. Selenium was administered along with methotrexate in order to evaluate the protective effects. The extent of hepatotoxicity and protective effects were analyzed by measuring levels of serum liver enzymes and histological examination of liver samples. Statistical analysis: One way ANOVA test was applied for comparison of biochemical markers between groups. Chi Square test was also applied for histopathological results. Results: In present study, results showed that significant hepatotoxicity was observed (p < 0.05) with elevated levels of serum liver enzymes with methotrexate. Selenium was found to significantly reduce the hepatotoxicity (p < 0.05). Histological examination revealed steatosis, which was found to be markedly decreased after treatment with selenium along with methotrexate. Conclusion: We concluded that selenium helped to protect against hepatotoxicity induced by methotrexate in mice. Keywords: Drug induced liver injury, Hepatotoxicity, Methotrexate and Selenium.