p38信号通路对缺氧下大鼠星形胶质细胞增殖凋亡的影响及机制研究

Mechanism of p38 signaling pathway on proliferation and apoptosis of astrocytes under hypoxia

目的探讨p38信号通路对缺氧下星形胶质细胞增殖凋亡的影响。方法从新生2 d的大鼠的脑组织分离原代星形胶质细胞,将细胞分为缺氧组、缺氧+p38抑制剂组和正常组,各组细胞培养12 h后,Western blot检测细胞中p38、p-p38蛋白表达;24 h后CCK8实验和流式细胞术分别检测细胞的增殖及凋亡情况,Western blot检测Bcl-2、Bax、Cleaved Caspase3蛋白表达。结果缺氧组p-p38蛋白表达显著高于正常组,而缺氧+SB203580组p-p38蛋白表达显著低于缺氧组(P<0.01);缺氧组细胞存活率及Bcl-2蛋白表达均显著低于正常组,细胞凋亡率及Bax、Cleaved Caspase3蛋白表达均显著高于正常组(P<0.01);缺氧+SB203580组细胞存活率及Bcl-2蛋白表达均显著高于缺氧组,细胞凋亡率及Bax、Cleaved Caspase3蛋白表达均显著低于缺氧组(P<0.01)。结论 p38信号通路的激活降低了缺氧下星形胶质细胞增殖并促进细胞的凋亡,而抑制p38信号通路可提高细胞的增殖及抑制细胞的凋亡。

Objective To investigate the effect of p38 signaling pathway on proliferation and apoptosis of astrocytes under hypoxia.Methods Primary astrocytes isolated from neonatal 2 day rat brain tissue,the cells were divided into hypoxia group,hypoxia group＋p38 inhibitor group and the normal group.The p38 and p-p38 protein expression were detected after the cells were cultured 12 h by Western blot;cells proliferation and apoptosis were detected after 24 h by CCK8 assay and flow cytometry;expression of Bcl-2,Bax,Cleaved Caspase3 protein were detected by Western blot.Results The expression of p-p38 protein in hypoxia group was significantly higher than that in normal group,while the expression of p-p38 protein in hypoxia group＋SB203580 group was significantly lower than that in hypoxia group （P＆lt;0.01）.Cell survival rate and Bcl-2 protein expression in hypoxia group were significantly lower than that in the normal group,the apoptosis rate and Bax,Cleaved Caspase3 protein expression were significantly higher than those of the normal group,respectively （P＆lt;0.01）.Cell survival rate and Bcl-2 protein expression in hypoxia＋SB203580 group were significantly higher than that in hypoxia group,the apoptosis rate and Bax,Cleaved Caspase3 protein expression were significantly lower than those in hypoxia group （P＆lt;0.01）.Conclusion The activation of p38 signaling pathway can decrease the proliferation and promote the apoptosis of astrocytes under hypoxia,and the inhibition of p38 signaling pathway can increase the proliferation and inhibit the apoptosis of cells.