The purinergic P2X7 receptor (P2X7R) can be activated by ATP and plays significant and complex roles in neuropathology. However, research is limited concerning the role of P2X7R in radial glia following hypoxia-ischemia (HI). In this study, radial glial clone L2.3 cells were cultured and subjected to oxygen-glucose deprivation (OGD) to generate an HI model in vitro. We found that HI decreased P2X7R expression in the L2.3 cells. Activation of P2X7R in L2.3 cells by 3'-O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzATP) led to cell death in a dose- and time-dependent manner, while a P2X7R antagonist, oxidized ATP (oATP), alleviated the injury induced by BzATP or HI. We also found that P2X7R modulated the phosphorylation of glycogen synthase kinase-3β (GSK-3β). The present findings suggest that L2.3 cells express P2X7R, and this receptor may be involved in HI injury of radial glia by mediating phosphorylation of GSK-3β.