Endothelial cell dysfunction is a fundamental injury of atherosclerosis cardiovascular disease (ASCVD), closely linked to ferroptosis, which is a novel type of cell death induced by iron-dependent lipid peroxidation. Several clinical trials have suggested that empagliflozin, a selective inhibitor of sodium glucose co-transporter 2, reduces the risk of hospitalization for heart failure and cardiovascular death in patients with type 2 diabetes. However, little is known about the mechanism of EMPA in endothelial cell ferroptosis in ASCVD. The aim of the present study was to evaluate the potential mechanism of EMPA against ferroptosis induced by RAS-selective lethal 3 (RSL3) in endothelial cells.EA.hy926 human umbilical vein endothelial cells were cultured in vitro and were divided into four groups: The Control, RSL3, RSL3 + low-concentration EMPA intervention and RSL3 + high-concentration EMPA intervention groups. Iron-dependent lipid peroxidation was assessed by detecting the fluorescence intensity of ferrous ion (Fe), lipid reactive oxygen species (ROS) and content of malondialdehyde (MDA). The expression of ferroptosis-related genes was assessed by RT-qPCR and western blotting. siRNA nuclear factor erythroid 2-related factor 2 (NRF2) was transfected into EA.hy926 cells to measure the expression of target genes.It was demonstrated that the level of MDA, Fe and lipid ROS was higher in the RSL3-treated group compared with the EMPA intervention group, while EMPA markedly mitigated that effect. In addition, EMPA can reverse the RSL3-induced low expression of glutathione peroxidase 4 (GPX4) and high expression of ACSL4 in endothelial cells, as evidenced by the upregulation of nuclear factor transcription factor nuclear factor, erythroid 2 (NFE2)-related factor 2 and heme oxygenase-1 expression, while siNRF2 transfection impaired the antiferroptosis effect of EMPA. The present study indicated that EMPA may inhibit RSL3-induced ferroptosis in endothelial cells by activating the NRF2/heme oxygenase 1 gene pathway.