As an active component in traditional Chinese herbal medicine, Tetramethylpyrazine (TMP) played a critical role in the treatment of cardiovascular, cerebrovascular, and eye diseases. In this study, the TMP has obtained from the Chinese herbal medicine Ligusticum chuanxiong and investigated the protective effects of TMP on oxidative damage in human LO2 cells with the treatment of different concentrations of TMP. The results showed that TMP reserved oxidative damage caused by hydrogen peroxide in LO2 cells. Further other investigations suggested that TMP protected LO2 cells from oxidative damage through elevating activities of antioxidant enzymes and glutathione levels. The TMP treatment promoted the nucleus translocation of the nuclear factor erythroid 2-related factor 2 (Nrf2) and further induced expression of antioxidant response element. Finally, it could be concluded that TMP treatment suppressed apoptosis induced by hydrogen peroxide in LO2 cells. Thus, the TMP might protect LO2 cells from oxidative damage through antioxidative and anti-apoptotic effects.