Objective To investigate the expression changes of Natural Killer Group 2 member D (NKG2D) signaling pathway in microglia of mice after oxygen glucose deprivation injury and its potential role in microglia inflammatory response. Methods A mouse neuronal HT22-oxygen glucose deprivation (OGD) Model was established (HT22-model group). After 19 hours of modeling, the cells were oxygenated and glycosylated, and normal conditioned media (Control) and OGD media (Model) were collected to stimulate mouse BV2 microglia cells. The cell viability of HT22 and BV2 cells was detected by CCK-8 assay. NKG2D receptor signaling pathway and inflammation mediators expression in BV2 cells were detected by qPCR. The content of inflammation mediators in supernatant of BV2 cell culture was determined by ELISA.Results The survival rate of HT22 neurons was significantly decreased by OGD (P＜0.01), but the cell viability of BV2 cells was not affected by OGD supernatant (P＞0.05). Compared with the Control group, the mRNA expression levels of NKG2D receptor, its H60 ligand and adaptor protein DAP10 were significantly increased (P＜0.01), but there were no significant changes in RAE-1 and MULT1 ligand (P＞0.05). Meanwhile, the mRNA level and the protein content of BV2 cells of tumor necrosis factor (TNF-α) in Model group were significantly increased (P＜0.01). Conclusion Oxygen glucose deprivation can induce the expression of H60/NKG2D, downstream effector DAP10 and inflammation mediators in BV2 cells. These results suggest that the NKG2D receptor signaling pathway may play an important role in the inflammatory response of microglia after cerebral ischemia.