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

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

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索(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(0.1～1.0μmol/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LpmRNA表达和蛋白分泌显著减少。结论0.1～1.0μmol/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。

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 umol/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 umol/L） can inhibit the adipogenic differentiation of BMSC by down-regulating the expression of PPARγ and FABP4.