全反式维甲酸抑制猪脂肪间充质干细胞的成脂分化

All Trans Retinoic Acid Inhibit Adipogenesis Differentiation of Porcine Adipose Mesenchymal Stem Cells

分离培养猪脂肪间充质干细胞（adipose mesenchymal stem cells，AMSCs），流式细胞仪鉴定其表面标记．利用MTT比色检测不同浓度的全反式维甲酸（all trans retinoic acid，ATRA）对猪AMSCs增殖的影响；光学显微镜下观察猪AMSCs向脂肪细胞分化的形态学变化；油红O染色提取法分析不同浓度ATRA对猪AMSCs成脂分化的影响；RT-PCR检测脂肪细胞分化标志基因驴己和aP2 mRNA的变化．MTT比色结果显示，生理浓度（1×10^-9～1×10^-8mol／L）和药理浓度（1×10^-7～1×10^-5mol／L）ATRA认对猪AMSCs增殖均没有影响．油红O染色提取法结果表明，除1×10^-7mol／LATRA对猪AMSCs成脂分化没有影响外，生理浓度（1×10^-9～1×10^-8mol／L）和其它药理浓度（1×10^-6～1×10^-5mol／L）ATRA均显著抑制猪AMSCs成脂分化（P〈0.05）．RT-PCR检测显示，ATRA显著抑制脂肪细胞分化标志基因LPL和aP2 mRNA表达（P〈0．05）．

All trans retinoic acid （ATRA） has widely biological effects, which can regulate the proliferation and differentiation of cells. The purpose of the experiment is to investigate the function of ATRA on the proliferation and adipogenesis differentiation of porcine adipose mesenchymal stem cells（ AMSCs）. The effects of ATRA on proliferation and adipogenesis differentiation of AMSCs were detected by MTr and quantitation of Oil Red O staining respectively; the morphologic changes of adipogenesis differentiation were analyzed by Oil Red O staining. The differentiation of AMSCs to adipocyte was also verified by expression of special adipogenic marker genes LPL and aP2 through RT-PCR. These results show that physiological concentrations（ 1 × 10^-9 ～1 × 10^-8 mol/L）and pharmacological concentrations（1 × 10^-7 ～ 1 × 10^-5 mol/L）ATRA have no effect on the proliferation of porcine AMSCs. These results of quantitation of Oil Red O staining＇show that physiological and pharmacological concentrations ATRA blocks the Adipo-genesis differentiation of porcine AMSCs significantly besides 1 × 10^-7 mol/L（ P 〈 0.05）. The result of RT-PCR show that ATRA significantly inhibits the expression of adipogenic marker genes LPL and aP2 （ P 〈 0.05）.