| 叶记林,于有江,吴爱莲,等.氧化应激在 TRAIL 诱导 Hela 细胞凋亡中的作用[J].安徽医药,2015,19(11):2090-2093. |
| 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 |
| 投稿时间：2015-04-11 |
| DOI： |
| 中文关键词: TRAIL Hela 细胞 细胞凋亡 氧化应激 |
| 英文关键词: TRAIL Hela cel1s apoptosis oxidative stress |
| 基金项目:江苏省卫生职业技术教育研究科研课题基金 (No J201311) |
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| 中文摘要: |
| 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 40 μg·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 40 μg·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。 |
| 英文摘要: |
| 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 Cin 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 Cto 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. |
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