Objective To explore the effect of electroacupuncture on CD34⁺VEGFR2⁺endothelial progenitor cell (EPC)-derived vessels and stromal cell-derived factor-1α (SDF-1α)/CXCR4, and study its mechanism of promoting angiogenesis in hippocampus after focal cerebral ischemia/reperfusion. Methods A total of 180 healthy male adult Sprague Dawley (SD) rats were randomly divided into sham operation (sham) group, model (I/R) group, electroacupuncture (I/RE) group, I/RE plus AMD3100 (A specific antagonist of CXCR4) group (I/REA) and AMD3100 (I/RA) group. The rats received filament occlusion of the right middle cerebral artery for 2 hours followed by reperfusion. Electroacupuncture was applied to "Baihui" (GV20)/"Siguan" (Hegu LI 4/Taichong LR 3) acupoints for 30 min, once a day. The mRNA expression of SDF-1α and CXCR4 were detected by reverse transcription-polymerase chain reaction (RT-PCR). Double immunofluorescence was used to stain CD34⁺VEGFR2⁺ EPC-derived vessels. Results Compared with the sham group, the mRNA expressions of SDF-1α and CXCR4 were significantly upregulated in I/R and I/RE group(P<0.05), but that in I/RE group was more significantly increased than I/R group(P<0.05). In addition, the mRNA expression of SDF-1α and CXCR4 were highly increased on day 1 in the I/REA group than that of I/RE group, but decreased than that of I/RE group on day 7 after reperfusion (P<0.01). CD34⁺VEGFR2⁺ EPCs-derived vessels were obviously increased on 3d and 7d in the I/RE group compared with that of the I/R group, and significantly decreased on 7d in the I/REA group compared with that of the I/RE group (P<0.01). Conclusions Electroacupuncture can effectively promote angiogenesis through upregulating the expression of SDF-1α and CXCR4 in rat ischemic hippocampus after focal cerebral ischemia/reperfusion.