氧化应激在TRAIL诱导Hela细胞凋亡中的作用  

Oxidative Stress Effect on the Apoptosis of Hela cells Induced by TRAIL

作　　者：叶记林[1] 于有江[1] 吴爱莲[1] 王冬燕[1] 彭建明[1] 刘永春[2] 刘延庆[3] 

机构地区：[1]扬州市职业大学医学院,江苏扬州225009 [2]江苏省苏北人民医院儿科,江苏扬州225000 [3]扬州大学医学院,江苏扬州225001

出　　处：《安徽医药》2015年第11期2090-2093,共4页Anhui Medical and Pharmaceutical Journal

基　　金：江苏省卫生职业技术教育研究科研课题基金(No J201311)

摘　　要：目的探讨氧化应激在肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor related apoptosis inducing ligand,TRAIL)诱导人宫颈癌Hela细胞凋亡中的作用。方法 40μg·L^(-1)TRAIL处理细胞后,用MTT法检测TRAIL对细胞增殖的抑制作用;用分光光度法检测细胞内还原型谷胱甘肽(GSH)、丙二醛(MDA)含量;流式细胞仪检测胞浆内的活性氧(ROS)水平;激光共聚焦测定细胞线粒体膜电位(ΔΨm);蛋白质印迹法检测Bcl-2、Cyt C、DR 4、DR 5蛋白量。结果 40μg·L^(-1)TRAIL对Hela细胞的生长具有显著的抑制作用;使抗氧化因子GSH含量明显下降,氧化应激产物ROS、MDA含量明显增多;到6 h时ΔΨm不断降低,能明显下调抗凋亡蛋白Bcl-2的表达,引起线粒体Cyt C向细胞质的释放;同时上调死亡受体DR 5的表达。而抗氧化剂N-乙酰半胱氨酸(NAC)能抑制TRAIL引起的这些变化。结论氧化应激在TRAIL诱导的Hela细胞凋亡中起着重要作用,可能与ROS激活线粒体途径和上调DR 5的表达密切相关。Objective To explore the oxidative stress effect on the apoptosis of Hela cells induced by TRAIL. Methods After treatment with 40 μg · L^-1 TRAIL,inhibition of the cell proliferation was measured by MTT assay. The levels of glutathione hormone（ GSH） and malondialdehyde（ MDA） were detected by spectro photometric assay. And the contents of cytosolic reactive oxygen species（ ROS） were measured by flow cytometry. Confocal laser microscopy was used to analyze the change of mitochondrial transmembrane potential（ ΔΨm）. Furthermore,the expressions of Bcl-2,DR 4 and DR 5,and the location of Cyt C in cells were tested by western blot. Results 40 μg·L^-1 TRAIL exhibited anti-proliferative activity observably on Hela cells. And TRAIL induced the decrease of antioxidant GSH and the increase of products of oxidative stress,such as ROS and MDA,significantly. The level of ΔΨm decreased continually after exposure to TRAIL for 6 h. Subsequently,decrease of anti-apoptosis protein Bcl-2,translocation of mitochondrial Cyt C to cytoplasm,and up-regulation of death receptor DR 5 were also detected. But the antioxidant NAC can block the changes induced by TRAIL. Conclusions Oxidative stress may play an important role in TRAIL induced apoptosis in Hela cells,and probably the apoptosis is mediated by ROS-activated mitochondrial pathways and up-regulation of DR 5.

关 键 词：TRAIL HELA细胞 细胞凋亡 氧化应激 

分 类 号：R73-36[医药卫生—肿瘤]