番茄红素对鱼藤酮所致PC12细胞线粒体损伤的影响

Effect of Lycopene on Mitochondrial Damage of PC12 Cells Induced by Rotenone

摘要:

目的: 研究番茄红素对鱼藤酮诱导PC12细胞线粒体损伤的保护作用。方法: 采用鱼藤酮(0.5 μmol·L^-1)诱导PC12细胞损伤模型,给予番茄红素(3,10,30 μmol·L^-1)预处理2 h后,加入鱼藤酮,使终浓度达到0.5 μmol·L^-1,培养24 h后,CCK-8法检测细胞存活率,倒置相差显微镜观察细胞形态改变、透射电镜观察细胞线粒体超微结构,罗丹明123染色检测线粒体膜电位,以观察番茄红素的保护作用。结果: CCK-8结果显示与正常对照组比较,模型组细胞存活率下降为70.34%±2.81%(P<0.05),与模型组比较,番茄红素(3,10,30 μmol·L^-1)预处理后其细胞存活率分别提高到83.09%±3.15%,87.24%±2.15%,89.17%±2.26%(P<0.05);倒置相差显微镜观察显示鱼藤酮可导致PC12细胞凋亡,经番茄红素处理后,模型细胞凋亡情况明显改善;透射电镜观察显示鱼藤酮可导致PC12细胞线粒体超微结构发生改变,经番茄红素处理后,模型细胞凋亡情况明显改善;流式细胞仪检测结果显示与正常对照组比较,模型组细胞线粒体膜电位平均荧光强度下降为151.63±12.25(P<0.05),经番茄红素(3,10,30 μmol·L^-1)预处理后线粒体膜电位平均荧光强度分别提高到202.24±26.28,226.21±9.71,238.83±10.29 (P<0.05)。结论: 番茄红素对鱼藤酮诱导的PC12细胞线粒体损伤有保护作用。

Abstract:

Objective: To investigate the protective effects of lycopene against rotenone-induced mitochondrial damage in PC12 cells. Method: All the experiments were carried out with PC12 in vitro,which were divided into 5 group,control,model(0.5 μmol·L^-1),lycopene(3,10,30 μmol·L^-1) preincubation.Cell viability was determined by CCK-8 assay;inverted phase contrast microscope was used to observe the cell morphological was;the ultrastructural changes of neuronal mitochondria were viewed under transmission electron microscope.The flow cytometry was used to detect the mitochondrial membrane potential(MMP). Result: CCK-8 assay showed that rotenone had cytotoxicity in PC12 cells,compared with the cells in the control group,cell viability was declined to 70.34%±2.81%(P<0.05), the pre-treatment of lycopene(3,10,30 μmol·L^-1) could significantly increase the PCI2 cell viability, cell viability was increased to 83.09%±3.15%,87.24%±2.15%,89.17%±2.26%(P<0.05). Compared with the cells in the model group,the number and morphology of neuronal mitochondria changed distinctly in the lycopene pretreated cells. Compared with the cells in the model group,the pre-treatment of lycopene(3,10,30 μmol·L^-1)increased mean fluorescence intensity of mitochondrial membrane potential to 202.24±26.28,226.21±9.71,238.83±10.29 (P<0.05). Conclusion: Lycopene can exert protective effects against rotenone-induced mitochondrial damage in PC12 cells.