【摘要】 目的研究重组人激活素A(activin A)调节人脂肪干细胞(human adipose stem cells,h ASCs)中内胚层特异性转录因子GATA4和FOXA2表达的机制。方法将h ASCs分为对照组、activin A组和activin A+SIS3组,分别给予相应处理。用免疫细胞化学法和高内涵分析系统测定各组细胞中磷酸化smad3蛋白(p-smad3)、限定性内胚层特异性转录因子GATA4和FOXA2的蛋白水平表达情况,用实时定量PCR法测定GATA4和FOXA2的mRNA水平表达情况。结果分组处理细胞30 min,activin A组细胞细胞核中p-smad3的表达显著高于对照组和activin A+SIS3组(P<0.05);分组处理细胞72 h,activin A组细胞中GATA4和FOXA2的mRNA和蛋白表达显著高于对照组和activin A+SIS3组(P<0.05)。结论 activin A可活化h ASCs细胞内的p-smad3信号,并促进限定性内胚层特异性转录因子GATA4和FOXA2的表达;p-smad3的特异性抑制剂SIS3可阻断activin A对h ASCs中p-smad3、GATA4和FOXA2的调节作用。activin A调节h ASCs中限定性内胚层特异性转录因子GATA4和FOXA2的表达依赖于p-smad3信号。更多还原

【Abstract】 Objective To investigate the mechanisms of endoderm specific transcription factors GATA4 and FOXA2 induction in human adipose stem cells regulated by activin A. Methods Human adipose stem cells( h ASCs) were divided into control group,activin A group and activin A + p-smad3 specific inhibitor SIS3 group. Real time PCR,immunofluorescence and high content screening were used to analyze the mRNA expression of GATA4 and FOXA2,and the protein expression of p-smad3,GATA4 and FOXA2,respectively. Results The expression of p-smad3 increased at 30 min in activin A group compared to control group and activin A + SIS3 group( P < 0. 05). The mRNA and protein expression of GATA4 and FOXA2 significantly increased at 72 h in activin A group compared to control group and activin A + SIS3 group( P < 0. 05). Conclusion Activin A can activate the p-smad3 signaling in h ASCs,and promote the expression of endoderm specific transcription factors,while p-smad3 specific inhibitor SIS3 can block the effect of activin A.The expression of endoderm specific transcription factors GATA4 and FOXA2 in h ASCs induced by activin A depends on p-smad3 signaling.更多还原