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目的 探讨过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor γ,PPAR γ)基因表达对骨髓基质细胞(marrow stroma cell,MSC)向心肌细胞分化的影响及其调控作用。方法 原代培养Balb/c纯系小鼠MSC,应用脂质体转染法将pEGFP-N1-PPARγ表达载体转入MSC中,CA18筛选。RT-PCR检测mRNA表达,Western blot法检测蛋白表达。结果 未分化的MSC中PPARγmRNA不表达,GATA4、MEF2C、Nkx2.5表达呈阳性。经pEGFP-N1-PPARγ2转染后,调控成心肌细胞方向分化的转录因子GATA4(0.172±0.034对0.053±0.022,P<0.01)、MEF2C(0.164±0.041对0)、Nkx2.5(0.156±0.029对0)mRNA表达明显下调。MSC向成脂肪细胞方向分化,而向成心肌细胞分化受阻。结论 PPARγ抑制MSC分化过程中GATA4、MEF2C、Nkx2.5 mRNA的表达,抑制MSC向成心肌细胞分化,并促进其向脂肪细胞分化。
Objective To investigate the effects of peroxisome proliferator-activated receptor γ (PPARγ) gene on the differentiation of marrow stromal cells (MSC) into cardiomyocytes. Methods Balb / c mouse MSCs were cultured in primary culture. The pEGFP-N1-PPARγ expression vector was transfected into MSC by lipofection method and screened by CA18. RT-PCR detection of mRNA expression, Western blot detection of protein expression. Results PPARγ mRNA was not expressed in undifferentiated MSC, but GATA4, MEF2C and Nkx2.5 were positive. The transcriptional factors GATA4 (0.172 ± 0.034 vs. 0.053 ± 0.022, P <0.01), MEF2C (0.164 ± 0.041 vs. 0), Nkx2.5 (0.156 ± 0.029) regulated by cardiomyocyte differentiation after transfection with pEGFP-N1-PPARγ2 0) mRNA expression was significantly down-regulated. MSCs differentiate into adipocytes and block the differentiation into cardiomyocytes. Conclusion PPARγ inhibits the expression of GATA4, MEF2C and Nkx2.5 mRNA during MSC differentiation, inhibits the differentiation of MSCs into cardiomyocytes and promotes their differentiation into adipocytes.