Aims The objective of this study was to investigate whether vagal nerve stimulation(VNS)promotes recruitment of c-Kit positive cells through paracrine release of vascular endothelial growth factor(VEGF)from endothelial cells in coronary vessels in infarcted hearts.Methods and results VNS and its neurotransmitter Acetylcholine(ACh)enhanced VEGF expression and release in endothelial cells via m/nACh-R/PI3K/Akt/Sp1 pathway.The conditioned medium from endothelial cells obviously increased c-Kit positive cells migration at least in part through VEGF/VEGFR pathway.In vivo,VNS-stimulated VEGF expression in infarcted hearts resulted in massive mobilization and homing of c-Kit positive cells,triggering more vascular tube formation.Moreover,these c-Kit positive cells involved in angiogenesis in vitro and in vivo,leading to a reduced infarction size.Functional studies showed that VNS stimulated extensive angiogenesis in infarcted hearts as indicated by the expression of vWF Ⅷ,CD31 and α-SMA,and improved the left ventricular performance,while blockade of VEGF by shRNA or its receptor by antagonist significantly diminished the beneficial effects of VNS on ventricular performance.Conclusion VNS and its neurotransmitter ACh promotes angiogenesis in infracted heart at least in part through VEGF/VEGFR-mediated activation of c-Kit positive cells.