Aims: The aim of the present study was to investigate how cytochrome P450 (CYP) 2J2-derived epoxyeicosatrienoic acids (EETs) regulate the AKT1 and FOXO1 and BIM pathway and protect against endothelial apoptosis in the development and progression of atherosclerosis. Methods and Results: Recombinant adeno-associated virus (rAAV)-mediated CYP2J2 overexpression increased EET levels and prevented high-fat diet-induced atherosclerosis in ApoE(-/-) mice, which was associated with reduced vascular apoptosis. We also observed that CYP2J2 overexpression suppressed the weight gain induced by a high-fat diet. In vitro CYP2J2 overexpression increased EET levels, subsequently preventing tumor necrosis factor-a-induced apoptosis in human umbilical vein endothelial cells (HUVECs), which contributed to the pathogenesis of atherosclerosis. We observed that AKT1 suppressed BIM expression and apoptosis by inhibiting the function of FOXO1 in cultured HUVECs. Furthermore, we found that CYP2J2 overexpression and its metabolic products, EETs, suppressed tumor necrosis factor-alpha-induced apoptosis through AKT1 and FOXO1 and BIM dependent signaling in HUVECs. Conclusions: In summary, we identified a novel molecular mechanism for CYP2J2-derived EETs to protect against atherosclerosis.