水溶性辅酶Q10对鱼藤酮致PC12细胞凋亡的保护作用及其机制

Protective effect of water soluble CoQ10 on rotenone-induced apoptosis in PC12 cells

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24h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PC12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 CoQ10 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.