The aim of the present study was to examine the cytoprotective effects of Schisandrae semen essential oil (SSeo), purified from Schisandrae fructus, against oxidative stress-induced cell damage in C2C12 myoblasts. SSeo attenuated hydrogen peroxide (H2O2)-induced growth inhibition and exhibited scavenging activity against the intracellular reactive oxygen species (ROS) that were induced by H2O2. SSeo also inhibited comet tail formation, chromatin condensation and phosphor-histone γH2A.X expression, suggesting that it prevents H2O2-induced cellular DNA damage and apoptotic cell death. Furthermore, SSeo significantly enhanced the expression of heme oxygenase-1 (HO‑1) associated with the induction of nuclear factor erythroid-2-related factor 2 (Nrf2) in a time- and concentration‑dependent manner. In addition, the protective effect of SSeo on H2O2‑induced C2C12 cell damage was significantly inhibited by zinc protoporphyrin IX, an HO‑1 competitive inhibitor, in C2C12 cells. These findings suggest that SSeo augments the cellular antioxidant defense capacity through intrinsic free radical scavenging activity and activation of the Nrf2/HO‑1 pathway, thereby protecting the C2C12 cells from H2O2‑induced oxidative cytotoxicity. As a result, SSeo may have therapeutic potential in the development of functional foods and as the raw material for medicines to protect against oxidative stress.