摘要
目的观察鱼藤酮对PC12细胞凋亡以及细胞外信号调节酶1和2(ERK1/2)蛋白表达的影响,以深入探讨ERK1/2蛋白表达与鱼藤酮诱导PC12细胞凋亡之间的关系。方法通过建立PC12细胞鱼藤酮染毒模型,光镜下观察鱼藤酮对PC12细胞形态的影响;流式细胞仪检测PC12细胞凋亡率的改变;Western蛋白印迹法和免疫组化法检测PC12细胞ERK1/2蛋白表达及定位情况。结果鱼藤酮处理24h后PC12细胞形态明显改变且凋亡率增加,20μmol/LMEK抑制剂PD98059预处理1h与单纯鱼藤酮染毒比较凋亡率没有显著性改变;磷酸化ERK1/2蛋白表达量随着鱼藤酮剂量的加大而增高,且活化的ERK1/2蛋白主要集中在胞浆内表达。结论鱼藤酮体外作用可以诱导PC12细胞凋亡以及ERK1/2蛋白磷酸化表达增强。
Objective To investigate the effects of rotenone on the apoptosis and expression of extracellular signal regulated kinases 1 and 2 （ERK1/2） proteins in PC12 cells in order to find out correlation between ERK1/2 and rotenone-induced apoptosis of PC12 cells. Methods The model of PC12 cells treated by rotenone was established. The morphological changes of PC12 cells induced by rotenone were observed by microscope, and cell apoptosis rates were detected by flow cytometry. The expression and localization of ERK1/2 were determined by Western blot and immunohistochemical methods. Results After treatment with rotenone for 24 hours, PC12 cells showed evident morphological changes and higher apoptosis rates, and the changes became more evident as a higher-dose rotenone was given. Pretreatment with MEK inhibitor PD98059（20 μmol/L） for an hour had not changed cell apoptosis rates significantly compared with rotenone intoxication merely. The phosphorylated ERK1/2 expressions were significantly up-regulated at protein levels and mostly located in cytoplasm. Conclusions Rotenone could induce apoptosis of PC12 cells, and the expression of phosphorylated ERK1/2 proteins is enhanced by activation of MAPK/ERK signal cascade.
出处
《疾病控制杂志》
2007年第1期40-43,共4页
Chinese Journal of Disease Control and Prevention
基金
国家自然科学基金资助项目(30400347)
