白头翁皂苷D诱导U87MG细胞凋亡

Pulsatilla saponin D induces cell apoptosis in the U87MG cells

目的探讨白头翁皂苷D对人脑胶质瘤细胞系U87的增殖抑制和促凋亡效应及其机制。方法用MTT法检测细胞增殖抑制作用,并求得IC25、IC50和IC75,按浓度和时间分组,Annexin-Ⅴ/PI双染、流式细胞术和TUNEL/PI染色检测凋亡,Hoechst33342染色、透射电子显微镜、DNA-ladder检测细胞系改变,细胞免疫染色和Western blot检测相关凋亡蛋白表达。结果白头翁皂苷D在14.016 nmol/mL下对U87MG抑制率为94.034%(P<0.05);处理组细胞形态呈凋亡改变,细胞超微结构中发现自噬小体;处理组caspase-3、8、Bax和FasL表达上调(P<0.05),Bcl-2表达下调(P<0.05)。结论白头翁皂苷D对U87MG细胞有呈时间和浓度依赖的增殖抑制作用,且通过FasL/Fas途径诱导其发生凋亡。

Objective To investigate the proliferation inbibition and apoptosis induction of the Pulsatilla saponin D on the human malignant glioma cell lines U87MG in vitro.Methods The proliferation-inhibiting effect and apoptosis of the U87MG cells treated with the different concentrations of the Pulsatilla Saponin D for different times in vitro were detected by MTT assay and the flow cytometry respectively.The cell apoptosis was examined by Hoshest 33342,the fluorescence fiber mirror,TUNEL/PI,DNA-ladder,and the apoptotic pathways were analysed by the Western blot analysis.Results The proliferation inhibition of U87MG by Pulsatilla saponin D 14.016 nmol/mL was 94.034％ （P ＜ 0.05）.The nuclear shape of the U87MG cells was changed after treatment with Pulsatilla saponin D,and stained by Hoechst 33342 in a time-and-dose dependent manner.The total protein was extracted from the U87MG cells and the expression of the caspase-3,8,Bax and Fas-L were increased,while the level of Bax expression reduced.The level of Bcl-2 changed unobviously.Conclusions Pulsatilla saponin D can inhibit the proliferation of human malignant glioma cell lines U87MG with a time-dose dependent manner and induce the apoptosis of U87MG cells through the Fas/Fas-L pathway.