【摘要】 旨为探明micro RNA-199a(mi R-199a)对脂肪细胞凋亡的影响及核转录因子κB(nuclear factorκB,NF-κB)在其中的作用。培养3T3-L1脂肪细胞,使用肿瘤坏死因子α(tumor necrosis factorα,TNFα)诱导细胞凋亡。在此基础上分别使用NF-κB阻断剂PDTC和mi R-199a mimic处理细胞,判断过表达mi R-199a对TNFα诱导的脂肪细胞凋亡的影响及NF-κB在其中的作用。使用流式细胞仪分析细胞凋亡率,试剂盒检测Caspase3/7酶活,双荧光素酶报告系统检测NF-κB活性,q RT-PCR检测mi R-199a的表达水平,Western blotting检测NF-κB相关蛋白变化。结果显示,TNFα可以诱导分化的脂肪细胞凋亡并同时激活NF-κB,激活的NF-κB在TNFα诱导的脂肪细胞凋亡中发挥负调控作用。在脂肪细胞过表达mi R-199a明显抑制NF-κB的激活进而显著促进TNFα诱导的凋亡。mi R-199a通过调控NF-κB活性参与TNFα诱导的脂肪细胞凋亡。更多还原

【Abstract】 The aim of this study is to explore the effects of Micro RNA-199a(mi R-199a)on adipocytes apoptosis and the role of nuclearfactor κB(NF-κB). Tumor necrosis factor α(TNFα)was used to induce the apoptosis of cultured 3T3-L1 adipocytes. Based on this work,adding PDTC of NF-κB blocker and mi R-199 a mimic,the effect of overexpression of mi R-199 a on TNFα-induced apoptosis and the role of NF-κB in it were determined. The flow cytometry was used to analyze the cell apoptosis,kit Caspase3/7 to detect the enzymatic activities,dualluciferase reporter assay to detect the activity of NF-κB,q RT-PCR to detect the expression of mi R-199 a,and Western blotting to detect thechanges of relevant proteins. As results showed,TNFα induced the apoptosis of differentiated adipocytes,concurrently activated NF-κB.The activated NF-κB played a negative regulator role in the apoptosis of adipocytes mediated by TNFα. Over-expressed mi R-199 a significantlysuppressed the activation of NF-κB,and then remarkably promoted the apoptosis induced by TNFα. Conclusively,mi R-199 a involves in theTNFα-induced apoptosis of adipocytes through regulating NF-κB activation.更多还原