天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (11): 1171-1174.doi: 10.11958/20170715

• 细胞与分子生物学 • 上一篇    下一篇

Spry1对脂肪细胞分化的调控作用

岑运珠，刘颖，杨威利，王宝利   

  1. 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）；2 天津医科大学口腔医院牙体牙髓科；3 天津医科大学基础医学院

  • 收稿日期:2017-06-20 修回日期:2017-08-20 出版日期:2017-11-15 发布日期:2017-11-15
  • 通讯作者: 王宝利 E-mail:bliwang72@aliyun.com

The impact of Spry1 on adipocyte differentiation

CEN Yun-zhu,LIU Ying,YANG Wei-li,WANG Bao-li   

  1. 1 Key Laboratory of Hormones and Development (Ministry of Health), Metabolic Diseases Hospital & Institute of Endocrinology, Tianjin Medical University, Tianjin 300070, China; 2 Department of Endodontics Dentistry, Hospital of Stomatology, Tianjin Medical University；3 Basic Medical College of Tianjin Medical University
  • Received:2017-06-20 Revised:2017-08-20 Published:2017-11-15 Online:2017-11-15
  • Contact: WANG Baoli E-mail:bliwang72@aliyun.com

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摘要： 目的 探讨利用 siRNA 下调骨髓基质细胞系 ST2 细胞中 Spry1 的内源性表达对脂肪细胞分化的调控作用。方法 设计 Spry1 的靶向 siRNA 作为实验组，以转染 Control siRNA 作为对照组。在 ST2 细胞中转染 Spry1siRNA 和 Control siRNA 并进行成脂诱导，应用 qRT-PCR 检测 2 组细胞中的 Spry1 和成脂特异性因子过氧化物酶体增殖物激活受体（PPAR）γ、CCAAT 增强子结合蛋白（C/EBP）α、脂肪细胞表征因子 FABP4、脂肪因子 adipsin 的mRNA 表达水平。脂肪细胞分化成熟后，进行油红 O 染色，显微镜下观察脂肪细胞的染色情况及 Spry1 siRNA 对ST2 细胞成脂分化的影响。运用异丙醇萃取染色的脂肪细胞中的油红 O 并测定波长在 520 nm 处的光密度（OD）值。结果 转染 Spry1 siRNA 于 ST2 细胞后，与转染 Control siRNA 的对照组相比，Spry1 基因的表达水平明显下调，
Spry1 siRNA 可抑制脂肪细胞的分化，油红 O 染色显示实验组的脂肪细胞明显减少且 OD 值低于对照组；转染 Spry1siRNA 实验组的成脂特异性因子 PPARγ、C/EBPα、FABP4、adipsin 的 mRNA 表达水平显著下调，差异有统计学意义（P<0.05）。结论 Spry1 siRNA 可有效抑制前体细胞向脂肪细胞分化。

关键词: 间质干细胞, 成脂分化, 脂细胞, RNA, 小分子干扰, Spry1

Abstract: Objective To investigate the effect of Spry1 on adipocyte differentiation from ST2 cells by using siRNA.Methods Synthesized siRNA targeting Spry1 was used as experimental group, and control siRNA was used as control group. Spry1 siRNA and control siRNA were transfected into ST2 cells, then treating with adipogenic medium to induce adipocyte differentiation. The mRNA expression levels of Spry1 and adipocyte differentiation-specific genes PPARγ(peroxisome proliferator-activated receptor gamma), C / EBP α (CCAAT enhancer binding protein α), FABP4 (adipocyte marker gene fatty acid binding protein 4) and adipsin were examined by quantitative real-time PCR. The mature adipocytes were stained with oil red O, the staining adipocytes were observed by microscope, then understanding the effect of Spry1 siRNA on adipocyte differentiation. In addition, oil red O of the staining adipocytes was extracted with isopropanol, optical density (OD) values of oil red O were measured at a wavelength of 520 nm. Results Spry1 siRNA was transfected into ST2 cells. Compared with control group, the mRNA expression level of Spry1 was significantly reduced. The number of differentiated adipocytes from ST2 cells was decreased after staining with oil red O. And the OD value was lower than that of control group. The mRNA expression levels of adipocyte differentiation-specific genes PPARγ, C / EBP α, FABP4 and adipsin were significantly reduced compared with those of control group (P<0.05). Conclusion Spry1 siRNA can
effectively suppresse adipogenic differentiation from progenitor cells.

Key words:  mesenchymal stem cells, adipogenesis, adipocytes, RNA, small interfering, Spry1

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