摘要
目的:对吉祥草乙醇提取物(Reineckia carnea ethanol extracts,RCEE)诱导人肺癌A549细胞凋亡及其机制做初步研究。方法:采用cell counting kit-8试剂盒测定细胞存活率,AnnexinⅤ-FITC/PI双标后流式细胞仪检测细胞凋亡率;分别采用FLUO-8 AM、JC-1染色标记后流式细胞仪测定细胞内钙离子浓度和细胞线粒体膜电位,化学发光法检测Casapse-3/7活性。结果:RCEE对A549细胞增值的抑制作用较强且呈量效关系,作用8 h时IC50值为48.14 mg·L-1;RCEE能诱导A549细胞凋亡并呈剂量依赖关系;RCEE作用后A549细胞内游离钙离子水平升高、线粒体膜电位下降、Caspase-3/7活性升高;细胞内钙离子螯合剂BAPTA-AM和Caspase广谱抑制剂Z-VAD-FMK可以明显抑制RCEE诱导的A549细胞凋亡。结论:RCEE能抑制A549细胞增殖并诱导细胞凋亡,其机制可能与细胞内钙超载、线粒体膜电位下降、Caspase级联酶激活有关。
OBJECTIVE To investigate Reineckia carnea ethanol extracts （RCEE） inducing A549 cell apoptosis and its mechanism. METHODS Cell counting kit-8 was used to determine cell survival rate. The rate of apoptosis, intracellular free Ca2 concentration and mitochondrial membrane potential were quantitated using armexin-V FITC/PI staining, FLUO-8 AM staining and JC-1 staining, respectively. Caspase-3/7 activity was determined by luminescence assay. RESULTS RCEE couldsuppress the proliferation of A549 cell in a dose-dependent manner , its ICs0 was 48. 14 mg.L-l at 8 h. The apoptosis could be induced by RCEE and its level increased following the augmentation of the drug concentration. RCEE induced intracellular free Ca2＋ concentration increased, mitochondrial membrane potential decreased and caspase-3/7 activated. Furthermore, pretreat ment with eitherZ-VAD-FMK or BAPTA-AM effectively attenuated apoptosis induced by RCEE. CONCLUSION RCEE can inhibit the proliferation and induce apoptosis of A549 cell lines in vitro. The mechanism of inducing apoptosis may be mediated by the overloadin of intracellular calcium ion, the decreased mitochondrial membrane notential and the caspase activated.
出处
《中国医院药学杂志》
CAS
CSCD
北大核心
2013年第19期1580-1584,共5页
Chinese Journal of Hospital Pharmacy
基金
江西省科技厅青年科学基金资助项目(编号:20132BAB215033)
江西省卫生厅中医药科研基金项目(编号:2012A132)
