Objective:To observe the effect of electroacupuncture(EA) on the expression of Nestin and Neuron specific nuclear protein antigen(neuron-specific protein,NeuN) at different time points after cerebral ischemia reperfusion injury.Methods:One hundred and fifty SD male rats were used to generate a one-side middle cerebral artery occlusion(MCAO) model using thread embolism method,then randomly divided them into the sham operation group,MCAO group,EA group,Nerve Growth Factor(NGF) group and EA+NGF group.The three benzene chloride tetrazolium(TTC) staining was used to detect the volume of cerebral infarction in the rats.Immunohistochemical staining was used to observe the expression of immune positive cell markers such as endogenous neural stem cell(NSC) marker protein Nestin,bromodeoxyuridine(Brdu) and NeuN on 1,7 and 14 days after cerebral ischemia in rats,and compared with the sham operation group.Results:TTC detection of the cerebral infarction volume showed that the nerve function injury score of the EA+ NGF group was significantly lower than MCAO group(P < 0.01).Ischemia led to severe loss of brain function,and EA+ NGF helped in the recovery of cerebral ischemia.Immunohistochemical results showed almost no expression of Nestin in the sham operation group;positive cells was expressed significantly higher Nestin on the 7th day(P < 0.05);then was peaked on the 14 th day.Nestin/BrdU labeled cells in the sham operation group showed minor expression,which was increased on the 7th day(P < 0.05) and was peaked on the 14 th day after ischemia.The differences were statistically significant on the 7th and 14 th day among the EA group,NGF group,EA+NGF group and MCAO group(P< 0.05).NeuN expression was higher in the sham operation group,and was increased on the 7th day after ischemia.NeuN/BrdU labeled cells showed higher expression in the sham operation group,the most obvious improvement was in the EA group,and the NGF and EA+NGF groups also showed significantly increased expression,while MCAO group showed the least.Conclusion:Protective effect of EA combined with NGF on cerebral ischemia reperfusion injury may induce nerve cell regeneration,accelerate proliferation of newborn cells,and promote differentiation of newborn cells,which is important for the recovery of nerve function.