Objective：To examine the protective effects of eryrthropoietin（EP） on neonatal SD rat cardiomyocytes injury induced by hypoxia／reoxygenation and explore its mechanisms involved. Methods：The cultured neonatal SD rat cardiomyocytes were divided into four groups：①hypoxia／reoxygenation group（H／R）：deoxygenation for 2 h and reoxygenation for 4 h；②EP group：EP（10 U／ml） was added 1 h before deoxygenation，then deoxygenation for 2 h and reoxygenation for 4 h；③Wortmannin+EP group（W+EP group）：Wortmannin（100 nmol／L）, a specific phosphatidylinositol-3-kinase（PI3-K） inhibitor，was added at 30 min before EP incubation；④Control group. Cell apoptosis were measured by flow cytometry.The expression of EP receptor and p-Akt／Akt level were detected by Western blot. Results：EP significantly decreased the apoptotic index induced by hypoxia／reoxygenation and strongly increased p-AKt／Akt level. However, pretreatment with Wortmannin attenuated the cytoprotective effect of EP. Conclusion：Myocardial death and apoptosis are involved in hypoxia／reoxygenation injury，and EP can protect neonatal rat cardiomyocytes from hypoxia／reoxygenation-induced apoptosis. The protective effects are partly associated with PI-3K／AKt pathway.