Abstract: Objective To investigate the effects of[Gly14]-Humanin(HNG) pretreatment on NF-κB p65 inhibition, inflammatory response and apoptosis in the rats with focal cerebral ischemia-reperfusion injury. Methods Eighty healthy male Sprague-Dawley rats were randomly divided into sham-operation, modelp, normal saline and HNG groups, with 20 rats in each group. A rat model of acute middle cerebral artery occlusion(MCAO) and reperfusion was established by suture embolism, relevant medicines(normal saline in the rats of the sham-operation group and normal saline group, HNG in the rats of the HNG group) were given to rats 3 days before operation respectively(iv, three times, qd). After cerebral ischemia 3 h and 24 h of reperfusion, neurological deficit score(NDS) were performed for each group. ELISA was respectively used to detect levels of NF-κB p65 and IFN-γ in cerebral tissue. HE staining is used to observe morphologic changes of neuron cells in ischemia region. TUNEL staining was used to detect the neuron apoptosis. Then the linear correlation analysis was used to determine the correlation between the levels of IFN-γ and NF-κB p65 in cerebral tissue with HNG group. Results Compared with sham-operation group, the NDS, the levels of NF-κB p65, IFN-γ and the number of neurocyte apoptosis were significantly increased in the other three groups(P<0.05); Compared with model group and normal saline group, the NDS, the levels of NF-κB p65, IFN-γ and the number of neurocyte apoptosis were significantly decreased in HNG group(P<0.05); The levels of IFN-γ and NF-κB p65 in HNG group were perfect positive correlations(r=0.739, P<0.001). Conclusion [Gly14]-Humanin can alleviate the inflammation in the process of the focal cerebral ischemia reperfusion, resistance the neurocyte apoptosis of ischemic area, thus mitigate the neurologic deficit. Its mechanism is probably through to down-regulating NF-κB p65 levels and then to reduce the release of IFN-γ.