鱼藤酮诱导PC12细胞自噬的初步研究

Preliminary Study of Autophagy Induced by Rotenone in PC12 Cells

目的研究鱼藤酮处理对PC12细胞自噬水平的影响。方法培养PC12细胞,分别用0、0.01、0.05、0.1、0.5、1、5、10μmol/L鱼藤酮处理细胞12h,以及1μmol/L鱼藤酮分别处理细胞0、2、6、12、18、24h,Western blot检测各组细胞内LC3-Ⅰ和LC3-Ⅱ含量,荧光显微镜检测GFP-LC3荧光斑点的数量,透射电镜观察细胞超微结构。结果随着鱼藤酮处理浓度升高,LC3-Ⅱ/LC3-Ⅰ比值和GFP-LC3荧光斑点数量均随之增加;随着鱼藤酮处理时间延长,GFP-LC3荧光斑点数量也随之增加,LC3-Ⅱ/LC3-Ⅰ比值先升高后降低,透射电镜显示细胞线粒体受损和细胞自噬水平增加。结论鱼藤酮对PC12细胞自噬的影响具有量效性和时效性。

Objective To investigate the effects on autophagy induced by rotenone in PC12 cells. Methods PC12 cells was treated with 0,0. 01,0. 05,0. 1,0. 5,1,5,10 vcmol/L rotenone for 12 hours and with 1 μmol/L rotenone for 0,2,6,12,18,24 hours respectively. Expressions of LC3-I and LC3- II were detected by Western blot and dots of GFP-LC3 were examined by fluorescence microscopy. Cell ultrastructures were observed under transmission electron microscope. Results With increasing concentration of rotenone, the ratios of LC3- II / LC3- I and the numbers of dots increased accordantly. With increasing time of rotenone, the numbers of dots increased accordantly while the ratios of LC3- II / LC3- I rose at first and fell afterwards. The mitochondria were obviously damaged and the level of autophagy was increased in PC12 cells. Conclusion There is a time based and quantitative efficiency in the effects on autophagy induced by rotenone in PC12 cells.