天然产物研究与开发 ›› 2020, Vol. 32 ›› Issue (4): 541-548.doi: 10.16333/j.1001-6880.2020.4.001

• 研究论文 •    下一篇

姜黄素通过促进ROS产生诱导人脑胶质瘤细胞凋亡

刘国安*,靳亚东,冉苗苗,李贵琛,杨麒励,丁兰   

  1. 西北师范大学生命科学学院,兰州730070
  • 出版日期:2020-04-28 发布日期:2020-06-05
  • 基金资助:
    国家自然科学基金(31660101);甘肃省自然科学基金(17JR5RA081)

Curcumin inducing apoptosis of U87 cells by promoting ROS production

LIU Guo-an*,JIN Ya-dong,RAN Miao-miao,LI Gui-chen,YANG Qi-li,DING Lan   

  1. Northwest Normal University Collage of Life Science,Lanzhou 730070,China
  • Online:2020-04-28 Published:2020-06-05

摘要: 本文用姜黄素处理人脑胶质瘤U87细胞,以CCK-8法检测姜黄素对细胞增殖的影响,在光学显微镜下观察姜黄素处理后的细胞形态学变化;酶联免疫法测定NADPH氧化酶的含量;用DCFH-DA荧光探针检测细胞ROS含量,并对氧化应激指标总氧化力(T-AOC)、丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽(GSH)进行了检测。用Annexinv/PI双染后流式细胞术检测和AO/EB荧光染色检测细胞凋亡,并通过免疫印迹法检测了凋亡相关蛋白信号通路。结果表明,姜黄素通过提高细胞NADPH氧化酶活性促进ROS产生,引起细胞内T-AOC降低,MDA含量升高,GSH含量下降,SOD活性升高,使细胞处于氧化胁迫,并可能通过ROS的升高触发细胞信号通路,下调NF-κB/p65蛋白的表达,最终通过凋亡执行分子Caspase-3促使细胞凋亡。

关键词: 姜黄素, 活性氧, 氧化应激, 细胞凋亡

Abstract:

Human glioma U87 cells were treated with curcumin and the effect on cell proliferation was assayed by CCK-8 method,the morphological changes were observed NADPH oxidase in cells were tested by ELISA.DCFH-DA fluorescence stain was used and the cells ROS content was detected.The oxidative stress indexes total antioxidant capacity(T-AOC),malondialdehyde(MDA),superoxide dismutase(SOD) and glutathione(GSH) were tested.Apoptosis was analyzed by AO/EB fluorescence staining and flowcytometry after double staining with Annexinv/PI.Finally,apoptosis-related signaling pathway protein was detected by immunoblotting.The results showed that curcumin promoted ROS production by increasing cell NADPH oxidase activity,which decreased intracellular T-AOC,increased MDA content,decreased GSH content,and increased SOD activity.These indicated curcmin induced oxidative stress in cells,and possibly increased ROS to trigger the cell signaling pathway by down-regulating the expression of NF-κB/p65 protein,and finally promoted apoptosis through the apoptosis-executing molecule Caspase-3.

Key words: curcumin, reactive oxygen species, oxidative stress, apoptosis

中图分类号:  R285.5