P2X7R antagonist inhibits primary neuronal apoptosis induced by oxygen and glucose deprivation through p38MAPK

Objective To investigate the anti-apoptotic effect of purinergic P2X7 receptor (P2X7R) antagonist through p38 mitogen-activated protein kinase (p38MAPK) in primary neuron of oxygen-glucose deprivation (OGD) model. Methods The primary neurons in the cerebral cortex of C57BL/6J fetal mice were cultured and the OGD model was established. The primary neurons were divided into control group and OGD 0.5, 1, 2, 3, and 6 h groups. The expression of P2X7R was detected by Western blot. Then the primary neurons were divided into control group, OGD group, OGD+inhibitor (Brilliant Blue G, BBG) or SB203580) group. The protein expression of P2X7R, phosphorylated p38MAPK, p38MAPK, and activated caspase-3 (cleaved caspase-3) were detected by Western blot, and apoptosis was detected by immunofluorescence staining. Results The results of Western blot showed that P2X7R expression in OGD 1, 2, 3 and 6 h of primary neurons increased (P<0.05), and the highest expression was at 3 h. Compared with the control group, the expression of P2X7R, P-P38MAPK and cleaved caspase-3 in OGD group increased significantly(P<0.05). Compared with OGD group, the expression of P2X7R, P-P38MAPK and cleaved caspase-3 in OGD+BBG group was decreased (P<0.05). Compared with OGD group, the expression of P-P38MAPK and cleaved caspase-3 in OGD+SB203580 group decreased (P<0.05). Immunofluorescence staining showed that compared with the control group, the apoptosis of neurons in OGD group increased significantly (P<0.05), while compared with OGD group, the apoptosis of neurons in OGD+BBG group decreased significantly (P<0.05). Compared with the control group, the phosphorylation level of p38MAPK in OGD group was significantly higher (P<0.05), and the phosphorylation level of p38MAPK in BBG and SB203580 pretreatment group was lower (P<0.05). Conclusions P2X7R antagonists play an anti-apoptotic role in primary neuronal OGD model by inhibiting p38MAPK phosphorylation.