In this study we investigated whether ethanol could play neuroprotective effects against ischemic brain injury and the related mechanism. Cresyl violet staining results demonstrated that moderate dose of ethanol administered intracerebroventricularly (i.c.v.) had neuroprotective effect against ischemia-reperfusion induced neuronal cell death. Ethanol also inhibited the phosphorylation of JNK3 induced by cerebral ischemia-reperfusion. Three separate drugs, NS3763 (the selective antagonist of GluR5), GluR5 antisense oligodeoxynucleotides (AS-ODNs) and Bicuculline (an antagonist of GABA receptors), were found to inhibit the neuroprotective effect of ethanol. Moreover, the GABA receptor agonist muscimol could attenuate the JNK3 phosphorylation. Taken together, the results suggest that during ischemia-reperfusion ethanol may activate presynaptic GluR5-KA and postsynaptic GABA receptors continuously, and the activation of GABA receptors inhibits the JNK3 signal pathway. The results show a novel potential mechanism underlying ethanol protective effects.