Aim To study the protective effect and mechanism of glucagon like peptide-1 (GLP-1) on hypoxia/reoxygenation (H/R) injury of H9c2 cardiomyocytes. Methods H9c2 cardiomyocytes were cultured and randomly divided into the control group, H/R group, and 1,5, 10 μmol/L GLP-1 group to verify the protective effect of GLP-1. Also, H9c2 cardiomyocytes were randomly divided into si-NC group, si-NC＋H/R group, si-NC＋H/R＋10 μmol/L GLP-1 group, and si-Nrf2＋H/R＋10 μmol/L GLP-1 group to verify the role of nuclear factor E2-related factor 2 (Nrf2) in GLP-1 alleviating cell injury. The contents of lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB), malondialdehyde (MDA) and total antioxidant capacity (T-AOC) and the expression levels of Nrf2 and heme oxygenase-1 (HO-1) were detected after 24 hours treatment. Results The contents of LDH, CK-MB and MDA in H/R group were higher than those in control group, and the contents of T-AOC and the expression levels of Nrf2 and HO-1 were lower than those in control group. The contents of LDH, CK-MB and MDA in different doses of GLP-1 groups were lower than those in H/R group, the contents of T-AOC and the expression levels of Nrf2 and HO-1 were higher than those in H/R group. After knocking down Nrf2 expression, the contents of LDH, CK-MB and MDA in si-Nrf2＋H/R＋10 μmol/L GLP-1 group were higher than those in si-NC＋H/R＋10 μmol/L GLP-1 group, the content of T-AOC and the expression levels of Nrf2 and HO-1 were lower than those in si-NC＋H/R＋10 μmol/L GLP-1 group. Conclusion GLP-1 can protect H9c2 cardiomyocytes from hypoxia/reoxygenation injury, and activation of Nrf2/HO-1 pathway is a possible molecular mechanism.