JNK信号传导通路在星形胶质细胞缺氧/复氧后损伤的作用

目的 建立SD大鼠星形胶质细胞（astrocyte,AC）缺氧/复氧损伤模型,探讨JNK信号转导通路与星形胶质细胞损伤的关系.方法 体外培养新生SD大鼠星形胶质细胞,传至第3代,细胞自然纯化,实验设正常对照组（N）、SP600125组（SP组,10 μmol /L）、缺氧/复氧组（H/R组）和缺氧/复氧＋SP600125组（H/R＋SP组）.分别利用MTT法、AnnexinV/碘化丙啶（propidium iodide,PI）双染法、WB法检测缺氧8 h,复氧4、6、12、24 h时细胞存活率、凋亡率、c-Jun氨基末端激酶（JNK）、磷酸化JNK（p-JNK）的变化.结果 ①缺氧/复氧后细胞活力出现明显下降,SP600125阻断JNK通路后细胞活力高于H/R组;②缺氧/复氧后细胞凋亡率增加,SP600125阻断JNK通路后能够减轻细胞凋亡.③各组AC的JNK水平无显著变化,缺氧/复氧干预后p-JNK表达上调,用SP600125阻断JNK通路后,H/R＋SP组较H/R组p-JNK水平降低.结论 JNK信号通路参与了星形胶质细胞缺氧/复氧损伤过程.

The role of JNK signaling pathway in astrocytes after hypoxic/reoxygenation injury

Objective To establish hypoxia/reoxygenatiou induced injury model and investigate the relationship be-tween JNK signaling pathway and the astrocytes （AC） impairment induced by hypoxia/reoxygenation in vitro. Methods The AC from the cortex of SD fetal rat were cuhured in vitro. When subcultured to the third generation, the cells natu-rally purified. The AC were divided into 4 groups： normal contrast （N） group, SP600125 （SP） group, hypoxic/reoxy- genation（H/R） group, and hypoxie/reoxygenation ＋ SP600125 （ H/R ＋ SP） group. After 8 h of hypoxia, it began to reox- ygenation. Each group received reoxygenation for 4 h, 6 h, 12 h, and 24 h. MTT was used to detect the cell activity, and the apoptos＇s situation was evaluated by flow cytometry after Annexin - V/PI double staining. Western - blot semi - quantitative analysis was adopted to detect the JNK and p - JNK expression. Results ①Hypoxia/reoxygenation reduced the AC activity significantly, cell viability was higher than the H/R group with SP600125 block JNK signaling pathway. ②Hypoxia/reoxygenation increased the rate of cell apoptosis, and the rate was reduced after SP600125 blocked JNK sig-naling pathway. ③No significant changes were observed in JNK level among the 4 groups, p - JNK expression increased after the intervention of hypoxia/reoxygenation, and the levels of p - JNK reduced compared with H/R group after SP600125 inhibited the JNK signaling pathway. Conclusion JNK signaling pathway is involved in the process of AC in hypoxia/reoxygenation injury.