JNK信号通路介导化学性缺氧对PC12细胞的损伤作用

目的探讨C-Jun蛋白氨基末端激酶(C-Jun N-termi-nal kinase,JNK)通路在化学性缺氧模拟剂氯化钴(CoCl2)对PC12细胞损伤中的作用。方法应用化学性低氧模拟剂氯化钴(CoCl2)处理PC12细胞以建立化学性缺氧损伤模型。应用CCK-8比色法检测细胞存活率;Hoechst33258核染色法观察细胞凋亡的形态学改变;JC-1染色荧光显微镜照像检测线粒体膜电位(MMP);Western blot法检测JNK蛋白的表达水平。结果 600μmol.L-1CoCl2作用PC12细胞不同时间(12～48 h)后,可时间依赖性地抑制PC12细胞的存活率;600μmol.L-1的CoCl2处理PC12细胞48 h时,可引起细胞出现核固缩等典型的凋亡特征;CoCl2能明显的降低PC12细胞的MMP;CoCl2能诱导JNK的磷酸化,特异性的JNK阻断剂SP600125可抑制CoCl2对PC12细胞的上述损伤作用。结论 CoCl2可引起PC12细胞损伤,此作用可能与其诱导JNK磷酸化有关。

JNK-mediated chemical hypoxia-induced PC12 cell injury

Aim To investigate the role of C-Jun N-terminal kinase(JNK) in PC12 cell injury induced by Cobalt chloride(CoCl2).Methods PC12 cells were treated with Cobalt chloride(CoCl2) to set up a chemical hypoxia-induced cellular injury model.Cell viability was tested by cell counter kit(CCK-8);Cell apoptosis was observed by Hoechst33258 staining and photofluorography;Mitochondrial membrane potential(MMP) was determined by JC-1 staining followed by photofluorography;Phosphorylation of JNK was measured by western blot assay.Results From 12 to 48 h,CoCl2 time-dependently inhibited cell viability in PC12 cells.Treatment with CoCl2 at 600 μmol·L-1 for 48 h induced PC12 cell apoptosis and decreased MMP.CoCl2 upregulated JNK phosphorylation.SP600125,a specific blocker of JNK significantly inhibited CoCl2-induced PC12 cell injury.Conclusion CoCl2 may induce PC12 cell injury.This effect may be associated with the CoCl2-induced activation of JNK.