Dehydroepiandrosterone (DHEA) has been proposed to regulate muscle dystrophy, while the underlying mechanisms for its protection against muscle atrophy are unknown. The present study was carried out to improve our understanding of the mechanism of DHEA's protective effect on muscle cells. We observed that DHEA significantly decreased the loss of cell death associated with H2O2-induced toxicity. Pretreating the muscle cells with DHEA led to a reduction of the intracellular accumulation of reactive oxygen species (ROS) in response to H2O2. In addition, DHEA reduced the H2O2-induced phosphorylation of ERK and p38 in a dose-dependent manner. Moreover, DHEA stimulated the activation of Nrf2, which led to the expression of an antioxidant response gene, HO-1. These results suggest that both antioxidants and anti-inflammatory properties mediate DHEA's effects for protection against muscle atrophy.