Objective: To investigate the effect of electroacupuncture (EA) promoting hippocampal neural stem cells (NSCs) proliferation in rats with focal cerebral ischemia- reperfusion (I/R) via Notch pathway and to clarify the possible mechanism of EA in treatment of cerebral ischemia. Method: Fifty- four male adult SD rats were randomly divided into the sham operation control group (SC group), the ischemia model group (IC group) and the electroacupuncture group (EA group). Middle cerebral artery occlusion (MCAO) was performed to establish the focal cerebral I/R injury model. Proliferation of hippocampal NSCs in cerebral I/R injured rats was determined by nestin immunohistochemical staining. The expressions of Notch1 and intracellular domain of Notch(NICD) were detected by Western blotting and RT- PCR. The serum level of vascular endothelial growth factor (VEGF) was measured using enzyme-linked immunosorbent assay (ELISA). Result: EA at Quchi (LI11) and Zusanli (ST36) acupoints alleviated neurological deficits significantly and promoted the proliferation of hippocampal NSCs (IC vs EA: 173.40±38.76 vs 246.80±47.73, P=0.028) in cerebral I/R injured rats; enhanced the expressions of Notch1 and NICD, crucial signaling molecules in Notch signaling pathway, and increased the secretion of VEGF. Conclusion: The up-regulatory effect of EA on Notch signaling pathway and neurotrophic factor secretion may result in the promotion of NSCs proliferation and consequently provide the therapeutic effect on cerebral ischemia.