Objective To explore the synergistic effects and related mechanisms of brain-derived neurotrophic factor (BDNF) on aerobic exercise-induced angiogenesis and improved cardiac function. Methods In the in vitro experiments, human umbilical vein endothelial cells (HUVECs) were exposed to laminar shear stress (LSS) of 12 dyn/cm2 to mimic the effects of exercise training on vascular tissue. non?circulating flow, circulating flow, circulating flow with BDNF (50ng/ml) patterns were used to produce LSS. BDNF/TrkB protein expression levels were examined, meanwhile phosphorylated TrkB and Akt protein levels were also observed under the intervention of LSS induced by the three different flow patterns respectively; HUVEC were divided into five groups: control group, non?circulating flow group, circulating flow group, BDNF group, circulating flow with BDNF group; after intervention for 6 h, the capacities of cell migration and tube formation were analysed. In the in vivo experiments, fifty-eight rats were assigned to Sham, myocardial infarction (MIC), myocardial infarction + BDNF (MICB), myocardial infarction + exercise (MIE) and myocardial infarction + exercise + BDNF (MIEB) groups, The exercise groups were subjected to 4 weeks of treadmill running; The MIEB group included rats subjected to exercise and daily tail intravenous injection of rhBDNF at a dose of 0.4μg/kg 10 minutes before exercise, the MICB group included rats subjected to the same dose of rhBDNF only; The rats underwent cardiac functional evaluations, prior to exercise training, at 1 week after cardiac surgery and following the 4?week period of exercise training; The blood vessel density in the surrounding area of myocardial infarction was detected by immunohistochemistry after the 4?week period of training. Results An increased BDNF protein expression, as well as phosphorylated of TrkB and Akt levels were observed in HUVECs exposed to LSS generated by circulating fluid. LSS elicited sustained increase BDNF level in the circulating medium. Circulating fluid with BDNF could increase the TrkB and Akt phosphorylated degree and improve the angiogenesis ability of HUVECs. In HUVECs exposed to LSS generated by non-circulating fluid, an increased BDNF protein expression were also observed, while the TrkB and its downstream Akt pathway were inactive. The in vivo experiments confirmed that BDNF administration in association with aerobic exercise increased myocardial angiogenesis and improved cardiac function more significantly compared with that in rats receiving only exercise intervention. Conclusions Exercise activates BDNF/TrkB axis and its downstream Akt pathway through LSS in a BDNF?dependent manner; exogenous supplementation of BDNF in association with aerobic exercise may enhance the pro-myocardial angiogenesis effect of exercise and further improve cardiac function.