Naringenin (NG), a flavanone extracted from various plants, has potent vasoprotective effects likely related to the induction of heme oxygenase-1 (HO-1). In the current study, we investigated mechanisms underlying the effect of NG on HO-1 expression and high glucose (HG)- or free fatty acids (FFA)-induced apoptosis in human umbilical vein endothelial cells (HUVECs). First, we found that HUVECs exposed to NG exhibited enhanced HO-1 expression in a concentration- and time-dependent manner. Moreover, HUVECs treated with NG exhibited activation of phosphoinositide 3 kinase (PI3K)/Akt, extracellular-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK). LY294002 (a PI3K inhibitor) and SP600125 (a JNK inhibitor) reduced NG-induced HO-1 expression, whereas BIRB796 (a p38 inhibitor) and PD98059 (an ERK inhibitor) had no effect. The cytoprotective effects of NG were correlated with activation of the transcription factor NF-E2-related factor 2 (Nrf2), a critical regulator of HO-1 expression. Indeed, the results of our experiments using LY294002 and SP600125 indicated that NG may stimulate Nrf2 through PI3K/Akt and JNK pathway activation. Moreover, treatment of HUVECs with Nrf2 siRNA decreased NG-induced HO-1 expression. Finally, pretreatment of HUVECs with NG remarkably reduced HG- or FFA-induced cell apoptosis, and this effect was greatly abrogated in the presence of SnPP (an HO-1 inhibitor). Above all, our data show that NG increased HO-1 expression and reduced HG- or FFA-induced cell apoptosis in HUVECs by upregulating PI3K, JNK, and Nrf2 pathways, which may confer an adaptive survival response in diabetes-induced vascular injury. •The present study provides the first experimental evidence that Naringenin attenuated high glucose or free fatty acids-mediated apoptosis dependent on HO-1 activation in human umbilical vascular endothelial cells.•Naringenin activated HO-1 through the PI3K and JNK/Nrf2 pathway in vascular endothelial cells.•Naringenin may be considered a potential endothelium protection agent when blood vessel exposure to high glucose environment.