论文部分内容阅读
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。
Objective To investigate the role of C-Jun N-termi-nal kinase (JNK) pathway in the injury of PC12 cells induced by chemical hypoxia mimic CoCl 2. Methods PC12 cells were treated with cobalt chloride (CoCl 2), a chemical hypoxia simulator, to establish a model of chemical hypoxia injury. The cell viability was detected by CCK-8 colorimetric assay; Morphological changes of apoptosis were observed by Hoechst33258 nuclear staining; Mitochondrial membrane potential (MMP) was detected by JC-1 staining; Western blot was used to detect the expression of JNK protein . Results PC12 cells were treated with 600μmol·L-1CoCl2 for a time-dependent manner (12-48 h), and the survival rate of PC12 cells was inhibited in a time-dependent manner. Treatment of PC12 cells with 600 μmol·L -1 CoCl2 for 48 h resulted in the appearance of nuclei CoCl2 can significantly reduce the MMP of PC12 cells; CoCl2 can induce the phosphorylation of JNK. Specific anti-JNK inhibitor SP600125 can inhibit the above-mentioned injury effect of CoCl2 on PC12 cells. Conclusion CoCl2 can induce PC12 cell injury, which may be related to its induction of JNK phosphorylation.