Cathepsin L在鱼藤酮诱导的PC12细胞凋亡中的作用  

The Role of Cathepsin L in Rotenone-induced PC12 Cells Apoptosis

作　　者：袁诗宇 徐舒青 宋云珍[1] 梁中琴[1] 

机构地区：[1]苏州大学药学院,江苏苏州215123

出　　处：《现代生物医学进展》2017年第14期2640-2645,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81072656;81102466;81373430)

摘　　要：目的:研究Egr-1对Cathepsin L的调控在鱼藤酮(Rotenone)诱导的多巴胺能神经元PC12凋亡的作用,初步探讨Egr-1与Cathepsin L的关系及机制。方法:常规培养PC12细胞,分别取1μM,2μM的Rotenone处理,用倒置显微镜观察细胞的形态变化;确定敏感性最强的浓度后再在不同的时间点下用Western blotting检测Egr-1、Cathepsin L蛋白表达情况;采用Egr-1si RNA转染PC12细胞,空载体siVector转染PC12细胞,Hoechst染色法检测细胞凋亡,Western blotting检测各处理组中Egr-1、Cathepsin L蛋白的表达情况。结果:Western blotting结果显示,经Rotenone刺激过的PC12细胞在2μM的浓度下最敏感,其Egr-1和Cathepsin L蛋白的表达量显著增加,且Egr-1在15 min就有明显的增加,而Cathepsin L在30 min才明显增加,说明Egr-1的确是出现在Cathepsin L蛋白的上游;Egr-1siRNA转染的PC12细胞的Cathepsin L表达量明显低于于空载体转染PC12细胞。结论:多巴胺能神经元PC12在Rotenone刺激下,细胞内Cathepsin L的表达与细胞内Egr-1蛋白水平有关,并且在抑制Egr-1的表达后,细胞内Cathepsin L的表达也相应的降低。所以我们得出Egr-1对Cathepsin L可能有调控作用,从而来调控多巴胺能神经元的凋亡。Objective： To research the regulation between Cathepsin L and Egr-1 in Rotenone-induced dopaminergic neurons PC12＇s apoptosis, and explore the relationship between Cathepsin L and Egr-1 and its mechanism. Methods： Conventional culture PC12 cells, and we observed cell morphology by inverted microscope. We took 1 μM and 2 μM Rotenone to treat PC12 cells and detect its sensitivity. Respectively we chose the concentration of the most sensitivity and then at different time points, we examined the expression level of Cathepsin L and Egr-1 in PC12 cells treated by Rotenone using Western blotting. PC12 cells were transfected with Egr-1 siRNA plasmid or control siRNA plasmid-A, and Hoechst detects cell apoptosis, and Western blotting detects each treatment group of Egr-1 and Cathepsin L protein expression intracellular. Results： The results showed that PC12 cells had the most sensitivity at the concentration of 2 μ Rotenone. ① Western blotting results suggested that after Rotenone stimulated PC12 cells, Egr-1 and Cathepsin L protein expression increased significantly, and Egr-1 increased significantly at 15 minutes, and Cathepsin L was increased significantly at 30 minutes, indicating that Egr-1 is indeed on the upstream of Cathepsin L protein. ②Cathepsin L expression of Egr-1 siRNA-transfected PC 12 cells was significantly lower than that in PC12 cells transfected with empty vector. Conclusion： After treated by Rotenone, Cathepsin L expression of intracellular cells is related with the level of Egr-1 protein, and the interference of the expression of Egr-1 protein could made the intracellular expression of Cathepsin L correspondingly reduced. We guessed that Egr-1 could regulate the expression of Cathepsin L and then regulate apoptosis of dopaminergic neurons.

关 键 词：组织蛋白酶L EGR-1 鱼藤酮 PC12细胞 帕金森综合症 

分 类 号：R-33[医药卫生] R742.5