›› 2010, Vol. 30 ›› Issue (3): 299-.

• 论著(基础研究) • 上一篇    下一篇

ATRA对人骨髓间充质干细胞成脂肪细胞分化及瘦素生成的影响

王 云, 梁 辉, 陶 荣   

  1. 上海交通大学 |医学院新华医院血液科, 上海 200092
  • 出版日期:2010-03-25 发布日期:2010-03-24
  • 通讯作者: 陶 荣, 电子信箱: hkutao@gmail.com。
  • 作者简介:王 云(1982—), 女, 住院医师, 硕士生;电子信箱: wangyunshj@126.com。
  • 基金资助:

    上海市自然科学基金(08ZR1414800)

Effects of ATRA on adipogenic differentiation and leptin production of human bone marrow mesenchymal stem cells

WANG Yun, LIANG Hui, TAO Rong   

  1. Department of Hematology, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, |Shanghai 200092, China
  • Online:2010-03-25 Published:2010-03-24
  • Supported by:

    Shanghai Natural Science Foundation, 08ZR1414800

摘要:

目的 探讨全反式维甲酸(ATRA)对人骨髓间充质干细胞(BMSC)成脂肪细胞分化及瘦素(Lp)生成的影响。方法 体外分离、培养和鉴定BMSC。在诱导培养液中加入不同浓度的ATRA进行成脂肪分化诱导,光学显微镜下油红O染色观察BMSC分化情况; RT-PCR和Western blotting测定细胞过氧化物酶体增殖激活性受体γ(PPARγ)和脂肪酸结合蛋白4 (FABP4)基因及相应蛋白表达;RT-PCR和ELISA法检测分化过程中细胞Lp基因表达及蛋白分泌水平的变化。结果 ATRA(0.1~1.0 μmol/L)作用后,BMSC向脂肪细胞分化呈现浓度依赖性抑制,PPARγ、FABP4 mRNA和蛋白表达均呈现一致性下降;诱导后细胞Lp mRNA表达和蛋白分泌显著减少。结论 0.1~1.0 μmol/L ATRA可抑制BMSC向脂肪细胞分化及其Lp生成,其机制可能与下调PPARγ和FABP4表达有关。

关键词: 骨髓间充质干细胞, 全反式维甲酸, 过氧化物酶体增殖激活性受体γ, 脂肪酸结合蛋白4, 瘦素

Abstract:

Objective To investigate the effects of all-trans retinoic acid (ATRA) on the adipogenic differentiation and leptin (Lp)production of human bone marrow mesenchymal stem cells (BMSCs). Methods Human BMSCs were isolated, incubated and identified. ATRA at various concentrations was added to the adipogenic induction medium. Adipogenic differentiation was quantified by microscopic examination with oil red O staining. The expression of peroxisome proliferator-activated receptor-γ  (PPARγ) and fatty acid binding protein-4 (FABP4) was analysed by RT-PCR and Western blotting, and the level of Lp was quantified by RT-PCR and ELISA. Results At the concentration of ATRA (0.1—1.0 μmol/L), adipogenic differentiation rates decreased in a dose-dependent manner. The mRNA and protein expression of PPARγ and FABP4 decreased concordantly. The mRNA expression and protein production of Lp significantly decreased after induction. Conclusion ATRA (0.1—1.0 μmol/L) can inhibit the adipogenic differentiation of BMSC by down-regulating the expression of PPARγ and FABP4.

Key words: bone marrow mesenchymal stem cell, all-trans retinoic acid, peroxisome proliferator-activated receptor-γ, fatty acid binding protein 4, leptin