〔Abstract〕 Objective To explore the role of glutamate transporter 2(EAAT2) in electroacupuncture preconditioning to reduce cerebral ischemia in mice. Methods 32 male C57 BL/6 mice were randomly divided into 4 groups(n=8): whole brain ischemia-reperfusion(BCCAO) group, electroacupuncture pretreatment(EA)+ BCCAO group, EAAT2 inhibitor dihydrokainate acid(DHK)+EA+BCCAO group, EAAT2 agonist ceftriaxone sodium(CXT)+ BCCAO group. In the BCCAO group, only the bilateral common carotid arteries were ligated. The DHK+EA+BCCAO group and the EA+BCCAO group were pre-treated with electroacupuncture once a day and 30 mins a time for 5 consecutive days. DHK 200 μg/kg was injected into the lateral ventricle before ischemia 1 h. In the CXT+BCCAO group, ceftriaxone sodium 600 mg/kg was injected intraperitoneally once a day for 5 consecutive days. After 24 h of preconditioning, global cerebral ischemia was performed for 20 mins. Neurobehavioral scores and HE staining were used to calculate apoptotic neurons in hippocampal CA1 area. Western blot was used to detect EAAT2 protein expression, and immunofluorescence double-label staining was used to detect EAAT2 and astrocyte activation markers(GFAP). Results Compared with the BCCAO group, the EA+BCCAO group and the CXT+BCCAO group had higher behavioral scores and decreased the number of apoptotic neurons(P<0.05). Compared with the EA+BCCAO group, the scores of neurobehavioral characteristics in the DHK+EA+BCCAO group decreased, and the number of apoptotic neurons increased(P<0.05). Western blot results: compared with the BCCAO group, the expression of EAAT2 protein in the EA+BCCAO group and the CXT+BCCAO group increased(P<0.05); compared with the EA+BCCAO group, the expression of EAAT2 protein in the DHK+EA+BCCAO group decreased(P<0.05). Immunofluorescence double-label staining: compared with the BCCAO group, the expression of EAAT2 and GFAP protein in the EA+BCCAO group and the CXT+BCCAO group increased(P<0.05), the expression of EAAT2 and GFAP protein in the DHK+EA+BCCAO group and the EA+BCCAO group decreased(P<0.05). Conclusion Electroacupuncture preconditioning can produce cerebral ischemic tolerance by regulating EAAT2 on astrocyte membrane.