Abstract: AIM: To explore the protective effect and mechanism of recombinant human erythropoietin (rhEPO) on cardiomyocytes against oxidative injury. METHODS: An experimental model of oxidative injury was established using cultured neonatal rat cardiomyocytes. The survival rate of cultured cardiomyocytes incubated in the presence or absence of rhEPO was examined using the MTT assay. Myocyte damages were estimated by release of lactate dehydrogenase (LDH) and cellular structure. Malondialdehyde (MDA) content was measured as an index of lipid peroxidation. The activity of Na+-K+-adenosine triphosphatase (Na+-K+-ATPase) and glutathione peroxidase (GSH-PX) was also assayed. RESULTS: The presence of rhEPO improved the survival of cultured cardiomyocytes. EPO treatment significantly decreased LDH leakage and MDA production and restored activities of Na+-K+-ATPase and GSH-PX. CONCLUSION: Pretreatment with rhEPO protects cardiomyocytes from induced injury. The beneficial effect may be related to its antioxidant properties and protection of Na+-K+-ATPase from oxidative injury.