转化生长因子β受体Ⅰ影响脂肪细胞分化的研究

目的探讨siRNA 下调转化生长因子β 受体Ⅰ(TGFBRⅠ)基因表达后对ST2 细胞向脂肪细胞分化的作用.方法设计并合成针对TGFBRⅠ的靶向siRNA 作为实验组,以转染Control siRNA 作为对照组.应用qRTPCR检测并比较转染ST2 细胞后各组TGFBRⅠ mRNA 的表达和脂肪细胞特异性转录因子CCAAT 增强子结合蛋白(C/EBP)α、过氧化物酶体增殖物激活受体(PPAR)γ、脂肪细胞表征因子FABP4 mRNA 的表达水平.诱导5 d 后对2组细胞进行油红O 染色,检测TGFBRⅠ siRNA 对ST2 细胞分化的影响.利用激光共聚焦显微镜观察脂肪细胞的染色情况并对细胞...

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Bibliographic Details
Published in天津医药 Vol. 44; no. 9; pp. 1062 - 1064
Main Author 杨永旭 陈棣 张欣 王宝利
Format Journal Article
LanguageChinese
Published 天津医科大学代谢病医院内分泌研究所 卫生部激素与发育重点实验室 邮编300070 2016
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ISSN0253-9896
DOI10.11958/20160429

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Summary:目的探讨siRNA 下调转化生长因子β 受体Ⅰ(TGFBRⅠ)基因表达后对ST2 细胞向脂肪细胞分化的作用.方法设计并合成针对TGFBRⅠ的靶向siRNA 作为实验组,以转染Control siRNA 作为对照组.应用qRTPCR检测并比较转染ST2 细胞后各组TGFBRⅠ mRNA 的表达和脂肪细胞特异性转录因子CCAAT 增强子结合蛋白(C/EBP)α、过氧化物酶体增殖物激活受体(PPAR)γ、脂肪细胞表征因子FABP4 mRNA 的表达水平.诱导5 d 后对2组细胞进行油红O 染色,检测TGFBRⅠ siRNA 对ST2 细胞分化的影响.利用激光共聚焦显微镜观察脂肪细胞的染色情况并对细胞进行拍照.另外,用异丙醇萃取出油红O,测定油红O 在波长520 nm 处的光密度(OD)值,并进行组间比较.结果转染TGFBRⅠ siRNA 后,ST2 细胞TGFBRⅠ基因的表达水平明显下调,TGFBRⅠ siRNA 能够促进脂肪细胞生成,增强C/EBPα、PPARγ 及FABP4 mRNA 表达;经成脂诱导5 d 后,转染TGFBRⅠ siRNA 的细胞产生的脂滴明显较Control siRNA 组多,且OD 值高于Control siRNA 组.结论TGFBRⅠ siRNA 能有效促进脂肪细胞的分化,提示TGFBRⅠ可能是前体细胞向脂肪细胞分化的重要调控因子.
Bibliography:adipocytes; cell differentiation; receptors, transforming growth factor beta; CCAAT-enhancer-binding pro?tein-alpha; RNA, small interfering; adipocyte characterization factor FABP4
YANG Yongxu, CHEN Di, ZHANG Xin, WANG Baoli(Key Laboratory of Hormones and Development (Ministry of Health), Metabolic Diseases Hospital & Institute of Endocrinology,Tianjin Medical University, Tianjin 300070, China)
Objective To investigate the effect of transforming growth factor β receptor type Ⅰ (TGFBRⅠ) on adipocytedifferentiation by using a small interference RNA (siRNA). Methods The siRNA targeting TGFBRⅠwas synthesized asexperimental group, and negative control siRNA was used as control group. The efficiency of TGFBRⅠ depletion and theexpression levels of adipocyte- specific transcription factors CCAAT enhancer binding protein α (C/EBPα),peroxisomeproliferator- activated receptor gamma (PPARγ) and adipocyte marker gene fatty acid binding protein 4 (FABP4) weredetected by quantitative real-time PCR. After treating with adipocy
ISSN:0253-9896
DOI:10.11958/20160429