天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (9): 1062-1064.doi: 10.11958/20160429

• 细胞与分子生物学 • 上一篇    下一篇

转化生长因子 β 受体Ⅰ影响脂肪细胞分化的研究

杨永旭， 陈棣， 张欣， 王宝利△   

  1. 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）


  • 收稿日期:2016-05-16 修回日期:2016-05-25 出版日期:2016-09-15 发布日期:2016-09-28
  • 作者简介:杨永旭 （1990）， 男， 硕士在读， 主要从事生物化学与分子生物学研究
  • 基金资助:
    国家自然科学基金资助项目 （81271977， 81472040）

The Effect of Transforming Growth Factor β Receptor type I on Adipocyte Differentiation

YANG Yongxu, CHEN Di, ZHANG Xin, WANG Baoli△   

  1. Key Laboratory of Hormones and Development (Ministry of Health), Metabolic Diseases Hospital & Institute of Endocrinology,Tianjin Medical University, Tianjin 300070, China
  • Received:2016-05-16 Revised:2016-05-25 Published:2016-09-15 Online:2016-09-28

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摘要： 摘要： 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。

关键词: 脂细胞, 细胞分化, 受体, 转化生长因子 β, CCAAT 增强子结合蛋白 α, RNA, 小分子干扰, 脂肪细胞表征因子 FABP4

Abstract: Abstract: Objective To investigate the effect of transforming growth factor β receptor type Ⅰ (TGFBRⅠ) on adipocyte differentiation by using a small interference RNA (siRNA). Methods The siRNA targeting TGFBRⅠwas synthesized as experimental group, and negative control siRNA was used as control group. The efficiency of TGFBR Ⅰ depletion and the expression levels of adipocyte- specific transcription factors CCAAT enhancer binding protein α (C/EBPα)， peroxisome proliferator- activated receptor gamma (PPARγ) and adipocyte marker gene fatty acid binding protein 4 (FABP4) were detected by quantitative real-time PCR. After treating with adipocyte differentiation agents for 5 days, the cells were stained with oil red O, and the staining of adipocyte was observed and photographed by laser confocal microscope. In addition, with isopropanol extracted oil red O, optical density values of oil red O were measured at a wavelength of 520, and which were compared between groups. Results After transfection of TGFBR Ⅰ siRNA, gene expression levels of TGFBR Ⅰ were significantly reduced in ST2 cells, the number of differentiated adipocytes was significantly increased, and the mRNA levels of adipocyte specific transcription factor C/EBP α and PPARγ and adipocyte marker gene FABP4 were enhanced compared with those of control group. After treating with adipocyte differentiation agents for 5 days， the number of lipid droplets of cells with transfection of TGFBRⅠsiRNA was increased than that of cells with transfection of control siRNA. The value of optical density was higher in cells with transfection of TGFBRⅠsiRNA than that of control siRNA group. Conclusion TGFBRⅠ siRNA can effectively facilitate adipocyte formation, which suggests that TGFBR Ⅰis an important regulator of adipogenic differentiation from prog

Key words: adipocytes, cell differentiation, receptors, transforming growth factor beta, CCAAT-enhancer-binding pro?
tein-alpha, RNA, small interfering, adipocyte characterization factor FABP4