ROS不参与Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid诱导的HepG2细胞凋亡  

ROS is not involved in induction of cell death by Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid in HepG2 cells

作　　者：李立[1] 吴科锋[1] 刘义[1] 吕应年[1] 龚先玲[1] 陈功[2] 赖宝山[2] 梁念慈[1,3] 

机构地区：[1]广东医学院广东天然药物研究与开发重点实验室,广东湛江524023 [2]香港中文大学韦尔斯亲王医院外科学系 [3]广东医学院生物化学与分子生物学研究所,广东湛江524023

出　　处：《中国中药杂志》2010年第10期1287-1291,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(3987099);香港特别行政区政府创新科技基金(GHP/022/06)

摘　　要：目的:确定活性氧(ROS)生成在Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid(5F)所诱导的HepG2细胞凋亡中的作用。方法:采用MTT分析检测5F对HepG2细胞增殖的影响,再以Hoechst/PI法分析凋亡细胞的形态变化。为了评价细胞内ROS水平,采用了GENMED试剂盒。HepG2细胞经5F处理24 h,或1 mmol.L-1GSH预处理1 h再经5F处理24 h,然后以Cell Death Detection ELISA试剂盒检测胞浆核小体片段。结果:通过细胞活性分析证实,5F对HepG2的细胞毒作用随着5F浓度升高而增强。而凋亡变化,如染色质浓缩,则被Hoechst/PI染色所证实。经5F处理的HepG2细胞内ROS生成减少被观察到。5F诱导的胞浆核小体片段并不由于GSH降低ROS介导的信号转导水平而发生变化。并且,顺铂(CDDP)诱导的ROS生成可被5F清除,同时5F还显示了与CDDP协同杀伤细胞作用。结论:5F不仅能通过非ROS依赖途径诱导细胞凋亡,还可缓解氧化应激。Objective： To identify the role of reactive oxygen species（ROS） formation on cell death induced by Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid（5F） in HepG2 cells.Method： MTT assay was used to determine the effect of 5F on proliferation of HepG2 cells,and apoptotic morphological changes were assessed using Hoechst/PI assay.To evaluate intracellular ROS levels,a GENMED kit was used.HepG2 cells were treated with 5F for 24 h or with 1 mmol·L-1 GSH for 1 h prior to treatment with 5F for 24 h,then cytoplasmic mono-and oligonucleosomes were assessed with Cell Death Detection ELISA kit.Result： The cytotoxicity of 5F on HepG2 cells was elevated with increasing 5F concentrations,as evidenced by the cell viability assay,and the apoptotic changes such as chromatin condensation were confirmed by Hoechst/PI staining.The decrease in ROS generation was observed in HepG2 cells following treatment with 5F.Cytoplasmic mono-and oligonucleosomes induced by 5F were not changed by decreasing basal level of ROS-mediated signaling with GSH.Farther more,induction of ROS production by cisplatinum（CDDP） was canceled by treatment with 5F and 5F revealed a additive effect to cell killing by CDDP.Conclusion： 5F can not only induce apoptosis through non-ROS-depandent pathway,and can abate oxidant stress.

关 键 词：半边旗 5F HEPG2细胞 细胞凋亡 活性氧 

分 类 号：R285[医药卫生—中药学]