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    张涛. P2X7R拮抗剂通过p38MAPK抑制氧糖剥夺诱导的原代神经元凋亡[J]. 徐州医科大学学报, 2020, 40(4): 235-239.
    引用本文: 张涛. P2X7R拮抗剂通过p38MAPK抑制氧糖剥夺诱导的原代神经元凋亡[J]. 徐州医科大学学报, 2020, 40(4): 235-239.
    shu
    P2X7R antagonist inhibits primary neuronal apoptosis induced by oxygen and glucose deprivation through p38MAPK[J]. Journal of Xuzhou Medical University, 2020, 40(4): 235-239.
    Citation: P2X7R antagonist inhibits primary neuronal apoptosis induced by oxygen and glucose deprivation through p38MAPK[J]. Journal of Xuzhou Medical University, 2020, 40(4): 235-239.
    shu

    P2X7R拮抗剂通过p38MAPK抑制氧糖剥夺诱导的原代神经元凋亡

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

      摘要
    • 摘要: 目的 探讨嘌呤能P2X7受体（Purinergic P2X7 receptor, P2X7R ）拮抗剂通过p38MAPK（p38 mitogen-activated protein kinase）在原代神经元氧糖剥夺（oxygen glucose deprivation, OGD）模型中发挥的抗凋亡作用。方法 培养C57BL/6J胎鼠大脑皮层原代神经元，构建OGD模型。将原代神经元分为对照组，OGD 0.5，1，2，3和6 h组。Western blot检测P2X7R的表达变化。再将原代神经元分为对照组，OGD组，OGD+抑制剂（BBG或SB203580）组。Western blot检测P2X7R，磷酸化的p38MAPK，p38MAPK和活化的半胱天冬酶-3（Cleaved caspase-3）的蛋白表达，免疫荧光染色检测神经元凋亡。结果 Western blot结果显示：与对照组相比，P2X7R在原代神经元OGD 1，2，3，6 h表达均增加（P<0.05），第3 h 表达最高。与对照组相比，OGD组P2X7R，p-p38MAPK和Cleaved caspase-3的表达均明显升高（p<0.05）；而与OGD组相比，OGD+BBG组P2X7R，p-p38MAPK和Cleaved caspase-3的表达均下降（p<0.05）。与OGD组比较，OGD+SB203580组p-p38MAPK和Cleaved caspase-3表达下调（p<0.05）。免疫荧光染色显示：与对照组相比，OGD组神经元凋亡明显增加（p<0.05）；而与OGD组相比，OGD+BBG组神经元凋亡明显减少（p<0.05）。荧光强度比值分析显示：与对照组相比，OGD组p38MAPK磷酸化水平明显升高（p<0.05），BBG和SB203580预处理后p38MAPK磷酸化水平降低（p<0.05）。结论 P2X7R拮抗剂通过抑制p38MAPK磷酸化在原代神经元OGD模型中发挥抗凋亡作用。

       

      Abstract: Objective: To investigate the anti-apoptotic effect of purinergic P2X7 receptor (P2X7R) antagonist through p38MAPK (p38 mitogen-activated protein kinase) in primary neuron of oxygen-glucose deprivation (OGD) model. Methods: The primary neurons in the cerebral cortex of C57BL/6J fetal rats were cultured and the OGD model was established. The primary neurons were divided into control group, OGD 0.5, 1, 2, 3 and 6 h groups. The expression of P2X7R was detected by Western blot. The primary neurons were divided into control group, OGD group and OGD+ inhibitor (BBG or SB203580) group. P2X7R, phosphorylated p38mapK, p38MAPK and activated caspase-3 (Cleaved caspase-3) protein expression were detected by Western blot, and neuronal apoptosis was detected by immunofluorescence staining . Results: The results of Western blot showed that compared with the control group, the expression of P2X7R in primary neurons OGD 1, 2, 3, 6 h was increased (P < 0 05), and the expression of P2X7R was the highest at 3 h. Compared with the control group, the expression of P2X7R, pp38MAPK and Cleaved caspase-3 in OGD group was significantly higher than that in OGD group, while the expression of P2X7R, pp38MAPK and Cleaved caspase-3 in OGD+BBG group was significantly lower than that in OGD+BBG group. Compared with OGD group, the expression of p-p38MAPK and Cleaved caspase-3 in OGD+SB203580 group was down-regulated. Immunofluorescence staining showed that neuronal apoptosis in OGD group was significantly higher than that in control group (p < 0 05), while that in OGD+BBG group was significantly lower than that in OGD group (p < 0 05). Fluorescence intensity ratio analysis showed that compared with the control group, the phosphorylation level of p38MAPK in OGD group was significantly higher than that in control group (p < 0.05and the level of p38MAPK phosphorylation decreased after SB203580 pretreatment (p < 0.05and p < 0.05respectively). Conclusion: P2X7R antagonists play an anti-apoptotic role in primary neuronal OGD model by inhibiting p38MAPK phosphorylation

       

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