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目的探讨鱼藤酮致大鼠多巴胺能嗜铬细胞瘤(PC12)细胞凋亡中前列腺凋亡反应蛋白(Par-4)表达变化,为帕金森病(PD)的基因治疗提供新的靶点。方法以1、5、10μmol/L的鱼藤酮分别作用于PC12细胞6、24 h,采用MTT比色法检测细胞存活率;采用Western blot法筛选出Par-4表达改变明显的鱼藤酮浓度和作用时间,给予信号转导抑制剂,检测Par-4表达量的变化。结果1、5、10μmol/L鱼藤酮干预后,PC12细胞出现不同程度的受毒形态改变,6 h后,10μmol/L组光密度比值显著升高(P=0.029 646);20μmol/L c-jun氨基末端激酶(JNK)信号通路抑制剂Ginestin预处理1 h,光密度比值与单纯鱼藤酮作用组比较显著降低(P=0.013 1);24 h后,PC12细胞的存活率降低显著,1、5μmol/L 2组光密度比值均升高,与对照组差异有统计学意义(P=0.018 461,P=0.043 415)。结论鱼藤酮可诱导PC12细胞Par-4凋亡蛋白表达升高,具有时间和浓度依赖性,JNK通路参与其信号转导途径。
Objective To investigate the changes of the expression of Par-4 in apoptosis induced by rotenone in rat dopaminergic pheochromocytoma (PC12), and to provide a new target for the gene therapy of Parkinson’s disease (PD). Methods Rat PC12 cells were treated with 1, 5 and 10 μmol / L rotenone for 6 and 24 h respectively. Cell viability was detected by MTT colorimetric assay. The concentration of rotenone and the effect of Par-4 expression were determined by Western blot. Signal transduction inhibitors were administered to detect changes in Par-4 expression. Results After exposure to 1,5 and 10 μmol / L rotenone for 24 h, the morphological changes of PC12 cells were observed. After 6 h, the optical density of 10 μmol / L group was significantly increased (P = 0.029 646) and 20 μmol / L c-jun After treated with Ginestin, an inhibitor of JNK signaling pathway for 1 h, the optical density ratio decreased significantly compared with the untreated group (P = 0.013 1). After 24 h, the survival rate of PC12 cells decreased significantly. The survival rate of 1,5 μmol / The optical density ratio of L 2 group was significantly higher than that of the control group (P = 0.018 461, P = 0.043 415). Conclusion Rotenone can induce the apoptosis protein expression of Par-4 in PC12 cells in a time-and concentration-dependent manner, and JNK pathway participates in its signal transduction pathway.