过表达MicroRNA-199a促进TNFα诱导的脂肪细胞凋亡

doi:10.13560/j.cnki.biotech.bull.1985.2017.03.026

生物技术通报 ›› 2017, Vol. 33 ›› Issue (3): 180-185.doi: 10.13560/j.cnki.biotech.bull.1985.2017.03.026

过表达MicroRNA-199a促进TNFα诱导的脂肪细胞凋亡

齐仁立^1,2, 王琪¹, 吴永江³, 王敬¹, 黄金秀^1,2, 杨飞云^1,2

1. 重庆市畜牧科学院,重庆 402460;
2. 农业部养猪科学重点实验室,重庆 402460,
3. 西南大学荣昌校区,重庆 402460

收稿日期:2016-06-28 出版日期:2017-03-26 发布日期:2017-03-07
作者简介:齐仁立,男,副研究员,研究方向：动物脂肪代谢的基因调控,E-mail：qirenli1982@163.com;王琪为共同第一作者
基金资助:
国家自然科学基金项目（31470117,31302055）,重庆市畜牧科学院基本科研业务费项目（14403）

Over-expression of MicroRNA-199a Intensifies the TNFα-induced Apoptosis of Adipocytes

QI Ren-li^1,2, WANG Qi¹, WU Yong-jiang³, WANG Jing¹, HUANG Jin-xiu^1,2, YANG Fei-yun^1,2

1.Chongqing Academy of Animal Sciences,Chongqing 402460;
2.Key Laboratory of Pig Industry Sciences,Ministry of Agriculture,Chongqing 402460;
3.Southwest University Rongchang Campus,Chongqing 402460

Received:2016-06-28 Published:2017-03-26 Online:2017-03-07

摘要/Abstract

摘要： 旨为探明microRNA-199a（miR-199a）对脂肪细胞凋亡的影响及核转录因子κB（nuclear factor κB,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 factor α,TNFα）诱导细胞凋亡。在此基础上分别使用NF-κB阻断剂PDTC和miR-199a mimic处理细胞,判断过表达miR-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,qRT-PCR 检测miR-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miR-199a明显抑制NF-κB的激活进而显著促进TNFα诱导的凋亡。miR-199a通过调控NF-κB活性参与TNFα诱导的脂肪细胞凋亡。

关键词: MiR-199a, 脂肪细胞, 凋亡, TNFa, NF-κB

Abstract: The aim of this study is to explore the effects of MicroRNA-199a（miR-199a）on adipocytes apoptosis and the role of nuclear factor 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 miR-199a mimic,the effect of overexpression of miR-199a 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,dual luciferase reporter assay to detect the activity of NF-κB,qRT-PCR to detect the expression of miR-199a,and Western blotting to detect the changes 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 miR-199a significantly suppressed the activation of NF-κB,and then remarkably promoted the apoptosis induced by TNFα. Conclusively,miR-199a involves in the TNFα-induced apoptosis of adipocytes through regulating NF-κB activation.

Key words: miR-199a, adipocyte, apoptosis, TNFα, NF-κB

齐仁立, 王琪, 吴永江, 王敬, 黄金秀, 杨飞云. 过表达MicroRNA-199a促进TNFα诱导的脂肪细胞凋亡[J]. 生物技术通报, 2017, 33(3): 180-185.

QI Ren-li, WANG Qi, WU Yong-jiang, WANG Jing, HUANG Jin-xiu, YANG Fei-yun. Over-expression of MicroRNA-199a Intensifies the TNFα-induced Apoptosis of Adipocytes[J]. Biotechnology Bulletin, 2017, 33(3): 180-185.

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过表达MicroRNA-199a促进TNFα诱导的脂肪细胞凋亡

Over-expression of MicroRNA-199a Intensifies the TNFα-induced Apoptosis of Adipocytes

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