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目的:研究知母水提液对Hela细胞线粒体功能、形态及生物学作用的影响。方法:本研究选用Hela细胞,分为空白对照组,知母高、中、低剂量组。采用四甲基罗丹明乙酯(TMRE)和Mito Tracker Orange CMTMRos染色检测线粒体膜电位变化,Mito Tracker Green FM染色流式细胞术检测线粒体总质量,线粒体膜蛋白TOM20免疫荧光染色观测线粒体形态改变,并对细胞内活性氧含量和细胞耗氧量进行检测,采用原位末端标记法(TUNEL)及Annexin V/PI染色流式细胞术检测细胞凋亡,蛋白印迹检测线粒体及凋亡相关蛋白的表达。结果:知母水提液在30min内即以浓度依赖方式显著降低Hela细胞线粒体膜电位(P<0.01),线粒体质量轻度下降,线粒体膜蛋白TOM20、细胞耗氧量均降低,显著升高细胞浆Endo G、Cyto C蛋白含量及细胞凋亡比率(P<0.01)。结论:知母水提液对Hela细胞线粒体功能和形态有显著影响,并可能经此诱导细胞凋亡。
Objective: To study the effects of aqueous extract of Zhimu on the function, morphology and biological function of mitochondria in Hela cells. Methods: Hela cells were used in this study, divided into blank control group, Anemarrhena as high, medium and low dose groups. The change of mitochondrial membrane potential was detected by TMRE and Mito Tracker Orange CMTMRos staining. The mitochondrial total mass was detected by Mito Tracker Green FM flow cytometry. The mitochondrial morphology was observed by immunofluorescence staining with mitochondrial membrane protein TOM20. The content of reactive oxygen species (ROS) and cell oxygen consumption in the cells were detected. TUNEL and Annexin V / PI staining were used to detect the cell apoptosis. Western blotting was used to detect the expression of mitochondria and apoptosis related proteins. Results: The water extract of Zhimu significantly reduced the mitochondrial membrane potential (P <0.01) in a concentration-dependent manner in 30 min, the mitochondrial mass decreased slightly, the mitochondrial membrane protein TOM20 and cell oxygen consumption decreased, Plasma Endo G, Cyto C protein content and apoptosis ratio (P <0.01). Conclusion: The aqueous extract of Zhimu has a significant effect on the function and morphology of mitochondria in Hela cells, and may induce apoptosis of Hela cells.