雷公藤红素对Hela细胞增殖和凋亡的影响

Effect of Celastrol on the Proliferation and Apoptosis of Hela Cell

研究雷公藤红素对人Hela细胞的增殖抑制和诱导凋亡作用以及对活性氧、线粒体膜电位和细胞内钙离子水平的影响。方法:用不同浓度的雷公藤红素处理Hela细胞,测定细胞生长曲线,CCK-8试剂盒检测细胞存活率;流式细胞仪检测细胞凋亡情况,检测细胞内活性氧、线粒体膜电位和钙离子浓度高低,原子力显微镜(AFM)对雷公藤红素处理Hela细胞前后细胞膜形貌及超微结构进行分析。结果:雷公藤红素能够显著抑制Hela细胞的增殖,对Hela细胞的半数抑制浓度为0.8μg/m L;随着雷公藤红素浓度上升,细胞内活性氧水平逐渐升高,线粒体膜电位逐渐下降,钙离子水平逐渐升高,2.5μg/m L雷公藤红素可使细胞的凋亡率达到83%;对细胞周期进行检测表明,雷公藤红素诱导细胞凋亡主要是使细胞阻滞在S期,共聚焦显微镜对细胞骨架和细胞核进行检测,雷公藤红素显著破坏细胞骨架,细胞核发生皱缩和断裂从而诱导细胞凋亡,AFM结果可以看到雷公藤红素使细胞膜发生皱缩、膜表面粒径变大从而影响细胞膜电位和钙离子稳定。结论:雷公藤红素诱导的Hela细胞凋亡与细胞内活性氧、线粒体膜电位以及钙离子水平有着紧密联系。

To examine the effects of celastrol on the proliferation and apoptosis of Hela cell, as well as its effects on reactive oxygen species（ROS）, mitochondrial transmembrane potential and intracellular free Ca2＋ concentration. Methods：Use different concentrations of celastrol to treat Hela cells, and describe the growth curve of the cells, test the survival rate of cells with the help of CCK-8 kit, examine the apoptosis, the ROS, the intracellular free Ca2＋ concentration and the mitochondrial transmembrane potential of cells by using flow cytometry, and analyze the differences in the epicyte appearance and ultrastructure of the cells by using AFM. Results：Celastrol is able to inhibit Hela cell proliferation obviously, and the inhibiting concentration of half of the cells is 0.8μg/ml; with the increase of celastrol concentration, the levels of ROS and the Ca2＋ concentration are rising, the mitochondrial transmembrane potential is lowering, and 2.5μg/ml celastrol concentration can cause an apoptosis rate of 83%; the test of cell cycle shows that the functioning of celastrol on the apoptosis of the cells is in S phase arrest, and the confocal microscopy indicates that the cytoskeletons are apparently damaged and the cell nucleuses shrank and collapsed, which cause the apoptosis of the cells; AFM results show that celastrol causes the shrink of epicyte and the increase of particle diameter which affect the stability of mitochondrial transmembrane potential and Ca2＋ concentration. Conclusions：The apoptosis of Hela cells induced by celastrol are closely related to the ROS, the mitochondrial transmembrane potential and Ca2＋ concentration.