目的 探讨水溶性辅酶 Q10(CoQ10)对鱼藤酮(Rot)诱导帕金森病(PD)细胞模型的保护作用及其可能的机制,为CoQ10 用于PD的治疗提供理论依据.方法 将对数生长期的PC12 细胞分为对照组、Rot组、CoQ10 组及 CoQ10治疗组;CCK8 检测细胞存活率;采用荧光探针2',7'-二氢二氯荧光黄双乙酸钠(DCFH-DA)使用荧光分光光度计检测细胞内活性氧(ROS)水平;Western blot检测CoQ10 对凋亡信号蛋白Bcl-2、Bax、Caspase-9、active Caspase-3 及凋亡诱导因子(AIF)表达的影响.结果 Rot处理 24 h后的 PC12 细胞存活率显著降低(P＜0.01)并与剂量呈负相关,Rot引起 ROS水平升高(P＜0.01),CoQ10 可改善 Rot 处理诱导的 PC12 细胞存活率降低(P＜0.01)及降低 ROS 水平(P＜0.01);Western blot实验表明CoQ10 可降低Rot引起的Caspase-9(P＜0.05)、active Caspase-3(P＜0.05)及Bax (P＜0.01)的表达增多,上调Bcl-2 的表达(P＜0.01);阻止 AIF向核内转移(P＜0.05).结论 水溶性CoQ10 对 Rot致PC1 2 细胞凋亡具有保护作用,机制可能是通过清除细胞内氧自由基改善细胞氧化应激状态,阻止 AIF 向核内转移,以及上调Bcl-2、下调Bax、同时降低 active Caspase-3 和Caspase-9 的表达而实现的.%Objective To investigate the protective effect and the underlying mechanism of water soluble coenzyme Q10 (CoQ10)against rotenone induced injury on PC12 cells model.Methods PC12 cells were cultured with rotenone,water-soluble CoQ1 0 was added to the culture media 3 hours prior to the rotenone incubation.We determined cell viability by CCK8;reactive oxygen species (ROS)was detected by spectrophotometer;and Bcl-2, Bax,active Caspase-3,Caspase-9 and apoptosis-inducing factor (AIF)were measured by Western blotting after 24-hour rotenone incubation.Results After the treatment by rotenone,cell viability decreased significantly (P＜0.01)and ROS level increased (P＜0.01).CoQ10 could improve PC12 cell viability (P＜0.01)and reduce the level of ROS (P＜0.01).Western blotting experiments showed that CoQ10 could reduce rotenone-induced Caspase-9 (P＜0.05),active Caspase-3 (P＜0.05)and Bax (P＜0.01)expressions,increase the expression of Bcl-2 (P＜0.01),and prevent nuclear translocation of AIF (P＜0.05).Conclusion CoQ10 has a protective effect on rotenone-induced apoptosis in PC12 cells,the mechanism of which may be through scavenging ROS in cells;decreasing caspase-9 ,active caspase-3 and Bax expressions;and increasing the expression of Bcl-2 ;and preventing AIF nuclear translocation.
展开▼
