鱼藤酮对PC12细胞及大鼠脑线粒体的损伤作用

Effects of rotenone on PC12 cells and mitochondria in rat brain

目的:在细胞水平、脑组织水平,探讨鱼藤酮对线粒体的影响及机制。方法:以PC12细胞为研究对象,经鱼藤酮处理后,使用细胞计数试剂盒(CCK-8)检测细胞活力,JC-1探针检测线粒体膜电位,DCFH-DA荧光探针检测细胞活性氧(ROS)水平,荧光素酶发光法检测线粒体ATP的含量。以大鼠脑线粒体为研究对象,通过Clark氧电极法检测鱼藤酮对线粒体呼吸功能的影响。结果:鱼藤酮不同程度的降低细胞存活率(P<0.05),2μM的鱼藤酮与PC12细胞共处理48 h,细胞活力下降为对照组的47.57%。2μM鱼藤酮使细胞内ROS增加、ATP含量减少、膜电位降低(P<0.05)。经鱼藤酮处理后,大鼠脑线粒体Ⅲ态和Ⅳ态呼吸速率及呼吸控制指数显著降低(P<0.05)。结论:鱼藤酮在细胞水平、脑组织水平,均诱导线粒体损伤。

Objective:To investigate the effect and mechanism of rotenone on mitochondria of cells and brain tissue.Methods:PC12 cells were treated with rotenone.The survival rate of PC12 cells was measured by CCK8.The intracellular ROS and mitochondrial membrane potential were detected by fluorescence probe.The mitochondrial ATP was detected by luciferase luminescence.Furthermore,the respiratory function of rat brain mitochondria was measured by Clark oxygen electrode.Results:Rotenone decreased the cell viability(P<0.05).The viability of PC12 cells treated with 2μM rotenone for 48 h decreased to 47.57%of that in the control group.Rotenone at 2μM increased ROS,decreased ATP and membrane potential(P<0.05).After rotenone treatment,V3,V4 and respiration control rate of rat brain mitochondria significantly decreased(P<0.05).Conclusion:Rotenone can induce mitochondrial damage on cells and brain tissue.