基础医学与临床 ›› 2014, Vol. 34 ›› Issue (5): 654-660.

• 研究论文 • 上一篇    下一篇

白头翁皂苷—D诱导U87MG细胞凋亡

李博1,王元刚1,李娟2,汤海峰3,程光1   

  1. 1. 第四军医大学西京医院神经外科
    2. 第四军医大学神经外科
    3. 第四军医大学药学系
  • 收稿日期:2014-02-25 修回日期:2014-03-13 出版日期:2014-05-05 发布日期:2014-04-28
  • 通讯作者: 程光 E-mail:chg16801@163.com
  • 基金资助:
    国家自然科学基金面上项目

Pulsatilla saponin D –induced cell apoptosis in the U87MG cells

  • Received:2014-02-25 Revised:2014-03-13 Online:2014-05-05 Published:2014-04-28

摘要: 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检测细胞核改变,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14.016nmol/mL下对U87MG抑制率为94.034%(n=6, 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。

关键词: 白头翁皂苷-D,增殖抑制,凋亡,细胞周期

Abstract: Objective To investigate the proliferation inbibition and apoptosis induction of the Pulsatilla saponin D on the human malignant glioma cell lines U87MG in vitro. Method 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,and the cell apoptosis were examined by Hoshest 33342、the Fluorescence fiber mirror、TUNEL/PI、DNA-ladder,and the apoptotic pathways were analysed by the Western blot analysis. Result The proliferation inhibition of the U87MG by the Pulsatilla saponin D were 94.034%(n=6, p<0.05). The nuclear shape of the U87MG cells was changed after being treated 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 showed to be increased, while the level of Bax expression reduced, and the level of Bcl-2 changed unobviously. Conclusion Pulsatilla saponin D can inhibit the proliferation of human malignant glioma cell lines U87MG at time-dose dependent manner and induce the apoptosis of U87MG cells through the Fas/Fas-L pathway.

Key words: Pulsatilla saponin D,proliferation inhibition, apoptosis, cell cycle

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