去甲斑蝥素诱导超氧阴离子的产生介导HeLa细胞凋亡  

Superoxide Anion Induced by NCTD Promoting Apoptosis in HeLa Cells

作　　者：董秀[1] 包红[1] 冯晓丹 王英[1] 韩兆丰[1] 李海波[1] DONG Xiu;BAO Hong;FENG Xiaodan;WANG Ying;HAN Zhaofeng;LI Haibo(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847

出　　处：《中华中医药学刊》2021年第8期97-100,共4页Chinese Archives of Traditional Chinese Medicine

基　　金：第四批全国中医(临床、基础)优秀人才研修项目(国中医药人教发[2017]24);辽宁省自然科学基金资助计划(2019-MS-232);辽宁省教育厅重点攻关项目(L202069)。

摘　　要：目的观察活性氧(reactive oxygen species, ROS)在去甲斑蝥素(norcantharidin, NCTD)诱导的人宫颈癌HeLa细胞凋亡中发挥的调节作用。方法体外培养HeLa细胞,甲基噻唑蓝比色法(MTT)检测NCTD的细胞毒性,流式细胞分析法检测ROS的产生,倒置显微镜和荧光显微镜观察细胞的形态学变化,免疫蛋白印迹法观察凋亡相关蛋白Procaspase 9、Caspase 3和ICAD的表达水平变化。结果 NCTD能够时间依赖性地诱导细胞ROS产生,NCTD产生的ROS主要以超氧阴离子的形式存在,60μmol/L的NCTD作用于HeLa细胞24 h后可使细胞皱缩、变圆,形成凋亡小体,活化Caspase-9和Caspase-3,并裂解ICAD,诱导细胞凋亡的发生。超氧阴离子的清除剂SOD可显著逆转NCTD诱导细胞生长抑制和细胞凋亡,明显影响NCTD诱导的Caspase-9蛋白的裂解,Caspase-3的剪切和ICAD的降解。结论 NCTD诱导超氧阴离子的产生介导HeLa细胞凋亡。Objective To observe the effect of reactive oxygen species(ROS) induced by norcantharidin(NCTD) on apoptosis in HeLa cells. Methods MTT assay was performed to assess the cytotoxicity of NCTD in vitro. The ROS generation was detected by flow cytometry. The morphological changes were examined under inverted microscope and fluorescence microscope. The expression levels of protein Procaspase 9, Caspase 3 and ICAD were examined by Western blot. Results NCTD promoted intracellular ROS production of dominant amount of superoxide anion. Remarkable morphologic changes were observed with treatment of 60 μmol/L NCTD for 24 h, including cell shrinkage, membrane blebbing and apoptotic bodies. NCTD induced activation of Caspase-9, Caspase-3, degradation of ICAD. Blocking ROS generation with superoxide dismutase(SOD) completely protected the cells from NCTD-induced apoptosis and reverse activation of Caspase-9, Caspase-3, degradation of ICAD. Conclusion The superoxide anion induced by NCTD impaired the apoptosis in HeLa cells.

关 键 词：去甲斑蝥素 活性氧 超氧阴离子 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]