摘要
目的:研究辛伐他汀诱导人红白血病细胞株K562细胞凋亡及细胞内活性氧与Ca2+水平的变化,以探讨凋亡机制。方法:20μmol.L-1辛伐他汀处理K562细胞,24h后光镜观察细胞形态;流式细胞术检测细胞凋亡率、活性氧和细胞内游离Ca2+水平。结果:20μmol.L-1辛伐他汀作用K562细胞48h后出现核固缩、核碎裂和凋亡小体等形态学改变;An-nexinV-FITC/PI检测细胞早期凋亡率,处理组凋亡率高于对照组,随药物作用时间延长逐渐增大,具有时间依赖性。荧光染料2′,7′-二氯荧光乙酰乙酸(2’,7’-dichlorofluoresceindiacetate,DCFH-DA)检测K562细胞内活性氧,不同时间处理组与对照组比较活性氧均升高,峰值时间为24h,与对照组比较发生显著改变。荧光染料Fluo-3AM检测K562细胞内游离Ca2+浓度,不同时间细胞内游离Ca2+浓度均升高,峰值时间为12h,与对照组比较发生显著变化。结论:辛伐他汀诱导K562细胞凋亡的可能机制是通过提高细胞内活性氧及游离Ca2+水平,从而导致细胞凋亡。
AIM： To investigate the changes of reactive oxygen species （ROS） and Ca^2＋ levels in K562 cells induced by simvastatin to predict the mechanism of apoptosis. METHODS： K562 ceils cultivated in routine method were treated with 20μmol· L^-1 simvastatin. The raorphology change was observed under light microscope after 24 h. Flow cytometry assay was used to detect the apoptotic ratio, the ROS and the Ca^2＋ levels. RESUILTS： 24 h after K562 cells had been treated with 20μmol· L^-1 simvastatin, there were some morphological changes of the cells such as karyopyknosis, nuclear fragmentation and apoptotic body emerged. AnnexinV-FITC/ PI was used to detect the apoptic ratio of K562 cells, the apoptotic ratio of treated groups was markedly increased compared with the control groups in a time-dependent manner. ROS, which was detected by 2＇, 7 ＇-dichloro fluorescein diacetate （DCFH-DA）, was markedly increased in treated groups compared with the control groups at different time points （ 12, 24, 48, 72 h）, and the level ROS got its peak time after 24 h. The Ca^2＋ , which was detected by Fluo-3AM, was markedly increased in treated groups compared with the control groups at different time points, and the level ROS got its peak time after 12 h. CONCLUSION： K562 cells can be induced to apoptosis by simvastatin. The potential mechanism of apoptosis might be related to the increase of ROS and Ca^2＋ level.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2006年第12期1364-1368,共5页
Chinese Journal of Clinical Pharmacology and Therapeutics
