TNFα对小鼠肾上腺皮质细胞系Y1增殖、凋亡的影响 (作者:___________单位: ___________邮编: ___________) 作者:方媛,夏海鸣,孙佳,黄培林 【摘要】目的:探讨肿瘤坏死因子α(TNFα)对小鼠肾上腺皮质细胞系Y1增殖、凋亡的影响。方法:不同浓度、不同作用时间的TNFα处理小鼠肾上腺皮质细胞系Y1;MTT法检测细胞的增殖;台盼蓝拒染法检测细胞增殖抑制;流式细胞仪检测细胞的凋亡水平;分光光度法检测细胞caspase8/3的变化。结果:不同浓度的TNFα处理Y1细胞不同时间后,细胞的增殖能力明显下降();细胞凋亡水平明显提高();细胞caspase8/3水平明显升高()。结论:TNFα可抑制Y1细胞增殖,并通过提高caspase8/3的活性而诱导及促进细胞凋亡。 【关键词】肿瘤坏死因子α; Y1细胞; 增殖; 凋亡; caspase8; caspase3 [Abstract] Objective: To investigate the effects of tumor necrosis factor α(TNFα) on proliferation and apoptosis of mouse adrenal cortex cell lines Y1. Methods: The mouse adrenal cortex cell line Y1 was treated with TNF α at different concentrations during different duration. The cells proliferation was tested by MTT assay and cytostasis was detectd by Trypan Blue Coloring Method. Flow cytometry was used to observe the cells apoptosis level. The changes of Caspase8/3 of the cells were detected by spectrophotometry. Results: After being treated with TNFα of different concentration during different duration, reproductive activities of Y1 cells were significantly inhibited(); the apoptosis and Caspase8/3 levels of Y1 cells were both remarkably highered(). Conclusion: TNFα can inhibit proliferation of Y1 cells, and induce and promote the apoptosis by increasing caspase8/3 activity. [Key words] tumor necrosis factor α; Y1 cells; proliferation; apoptosis; caspase8; caspase3 肿瘤坏死因子α(TNFα)是一种主要由单核巨噬细胞产生的单核因子,具有选择性地杀伤某些哺乳类动物的肿瘤细胞,并通过半胱天冬酶级联介导和活化激活蛋白1、核因子κB两个转录因子的通路诱导其凋亡。近些年来,TNFα对内分泌细胞的作用逐渐受到关注。2007年,Mikhaylova等[1]研究发现,TNFα可以降低人类肾上腺皮质NCIH295R细胞的增殖活性并诱导其凋亡。但是,它诱导内分泌细胞凋亡的机制目前尚不明确,而且不同细胞对TNFα的敏感性亦不相同。本研究旨在探讨TNF α对小鼠肾上腺皮质细胞增殖、细胞凋亡及其可能机制,为今后深入了解TNFα介导的炎症反应对小鼠肾上腺皮质功能的影响提供实验基础。 1 材料和方法 材料 小鼠肾上腺皮质细胞系Y1(中国科学院上海细胞生物研究所);肿瘤坏死因子α(TNFα)(Peprotech公司);F12培养基(GIBCO公司)、胎牛血清(杭州四季青公司)、MTT(GIBCO公司)、DMSO(Sigma公司);台盼蓝染液(南京生兴生物技术有限公司);Annexin VFITC及PI细胞凋亡检测试剂盒(南京凯基生物公司);caspase8/3分光光度法检测试剂盒(南京凯基生物公司。 方法 细胞培养小鼠肾上腺皮质细胞系Y1用含10%的胎牛血清、 g NaHC
