【摘要】 目的研究小鼠成纤维细胞重编程为心肌样细胞(induced cardiomyocytes,iCMs)过程中淫羊藿苷(icariin,ICA)的作用及心血管发育中胚层相关蛋白1(mesoderm posterior 1,Mesp1)的表达。方法 MTT法筛选ICA的最佳药物浓度。GMT转染小鼠皮肤成纤维细胞,建立重编程心肌样细胞模型。qRT-PCR检测转染后重编程细胞中肌球蛋白重链6(myosin heavy chain 6, Myh6)的mRNA表达,免疫荧光细胞化学方法检测心肌肌钙蛋白I (cardiac troponinI, cTnI)及Mesp1的表达情况。结果 ICA最佳药物浓度为10-7mol/L。ICA组诱导iCMs高表达cTnI,是GMT组的1.3倍。Myh6 mRNA在重编程第3周达到高峰,且ICA组显著高于GMT组(P<0.01)。Mesp1与c TnI的表达趋势正好相反。结论 ICA可以提高成纤维细胞重编程为心肌样细胞的效率;Mesp1参与促进成纤维细胞重编程的过程。更多还原

【Abstract】 Objective To study the effect of icariin(ICA) and the expression of mesoderm posterior 1(Mesp1) in the reprogramming process of mouse fibroblasts into induced cardiomyocytes(iCMs). Methods The optimal drug concentration of ICA was screened by MTT assay. The GMT recombinant virus was used to infect mice skin fibroblasts and establishe a reprogrammed cardiomyocyte model. The expression of myosin heavy chain 6(Myh6) mRNA in reprogrammed cells at 1, 2, 3, and 4 weeks after transfection was detected by qRT-PCR, and the expression of cardiac troponin I(cTnI) and Mesp1 protein during the process reprogramming was detected by immunofluorescence. Results The optimal drug concentration for screening ICA was 10-7 mol/L. The induced cardiomyocytes(iCMs) with ICA was highly expressed cTnI protein, with 1.3 times higher than in GMT group. The expression of Myh6 mRNA reached a peak in the third week of reprogramming, and was significantly higher in ICA group than in GMT group. The expression of Mesp1 and the myocardialspecific marker cTnI was reversed. Conclusion ICA improved the efficiency of myocardial cell reprogramming, Mesp1 is involved in the process of promoting fibroblast reprogramming.更多还原