薯蓣皂苷元对鱼藤酮损伤PC-12细胞的保护作用研究

Effects of diosgenin on PC-12 cells damaged by rotenone

中文关键词:  薯蓣皂苷元  细胞自噬  帕金森病  鱼藤酮

英文关键词:diosgenin  autophagy  Parkinson's disease  rotenone

中文摘要:

      目的 研究薯蓣皂苷元对PC-12细胞生长的影响及其自噬诱导作用。方法 用薯蓣皂苷元(diosgenin, DG)干预PC-12细胞, 四甲基偶氮唑盐(methyl thiazolyl tetrazolium, MTT)法和平板克隆形成实验检测DG对PC-12细胞的保护作用。DG处理PC-12细胞, Western Blot方法检测P62、LC3-II蛋白的相对表达量。DG处理经过鱼藤酮诱导损伤的PC-12细胞, 激光共聚焦显微镜检测细胞内活性氧的变化, Mito Tracker Red方法观察线粒体变化。结果 MTT法和平板克隆形成实验结果表明, 薯蓣皂苷元在15 μg/mL时对PC-12细胞有保护作用。DG干预PC-12细胞12 h后, P62蛋白表达减少(P<0.05), LC3-II蛋白水平升高(P<0.05)。DG干预鱼藤酮损伤的PC-12细胞24 h后, 细胞内活性氧降低(P<0.001), 线粒体损伤减弱。结论 薯蓣皂苷元可以通过诱导细胞自噬对鱼藤酮诱导的PC-12细胞损伤发挥保护作用。

英文摘要:

      Objective To investigate the effects of diosgenin on the growth of PC-12 cells and its autophagy induction. Methods Diosgenin (DG) was used to intervene PC-12 cells, and the protective effects of DG on PC-12 cells were detected by methyl thiazolyl tetrazolium (MTT) assay and plate clone formation test. PC-12 cells were treated with DG, and the relative expression levels of P62 and LC3-II proteins were detected by Western Blot. The PC-12 cells damaged by rotenone were treated with DG, and the changes of intracellular reactive oxygen species (ROS) were detected by laser scanning confocal microscope and mitochondrial changes were observed by Mito Tracker Red method. Results The results of MTT assay and plate clone formation experiments showed that diosgenin protected PC-12 cells at 15 μg/mL. After 12 h of DG intervention in PC-12 cells, the expression of P62 protein was decreased (P<0.05), and the level of LC3-II protein was increased (P<0.05). After 24 h of DG intervention in rotenone-injured PC-12 cells, intracellular ROS was decreased (P<0.001), and mitochondrial damage was weakened. Conclusion Diosgenin could protect rotenone-induced PC-12 cell injury by inducing autophagy.