Aim To explore the effect of brain-derived neurotrophic factor (BDNF) on Sestrin2 expression and angiogenesis-related mechanisms in endothelial cells. Methods Human umbilical vein endothelial cells (HUVEC) were treated with BDNF (100 μg/L) for 1 h, 2 h, 4 h, 6 h, 8 h, and the protein and mRNA expression of Sestrin2 were detected with immunofluorescent staining, Western blot and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. HUVEC were divided into six groups:control group, BDNF (100 μg/L) group, BDNF+TrkB-Fc (1 mg/L) group, BDNF+KT-5823 (500 nmol/L) group, BDNF+L-NAME(NG-nitro-L-arginine methyl ester) (10－4 mol/L) group, BDNF+DMSO(dimethyl sulfoxide)group; after intervention for 4 h, the expression of Sestrin2 was detected with Western blot. HUVEC were divided into four groups:control group, BDNF (100 μg/L) group, BDNF+Sestrin2 siRNA group, BDNF+control siRNA group; after intervention for 6 h, the capacities of cell migration and tube formation were analysed.Results Sestrin2 mRNA increased in 2 h, 4 h, 6 h group compared with that of the 0 h, 1 h group (P＜0.001), while the protein expression of Sestrin2 increased in 2 h, 4 h, 8 h group compared with that of the 0 h, 1 h group (P＜0.05). BDNF-induced increase in Sestrin2 expression was abolished by L-NAME and PKG inhibitor (P＜0.001). BDNF-induced cell migration and tube formation were completely blocked because of the suppressed expression of Sestrin2 by Sestrin2 siRNA (P＜0.01). Conclusion BDNF confers certain aspects of its proangiogenic capacity through NO/PKG/Sestrin2 pathway.