p53 modRNA促进人诱导多能干细胞来源早期心肌细胞分化成熟的初步研究

p53 modRNA promoting the maturation of hiPSC-derived early cardiomyocytes

目的探索p53 modRNA对促进人诱导多能干细胞来源早期心肌细胞分化成熟的影响。方法将p53 modRNA转染至人诱导多能干细胞来源的早期心肌细胞,利用qRT-PCR检测心肌细胞成熟相关基因mRNA的表达情况;利用Western blot检测心肌细胞成熟相关蛋白的表达情况;利用细胞免疫荧光染色观察心肌细胞的形态、肌节发育情况。结果qRT-PCR结果表明,p53 modRNA处理过后的心肌细胞增殖相关基因表达减少,脂代谢基因表达水平增高,且心肌特异性基因表达量增高,提示心肌细胞内结构可能更加成熟。Western blot结果显示,转染组cTnI蛋白表达量相比未转染组有增高趋势。免疫荧光染色结果显示,与未转染组相比,转染组细胞形态趋近长棒型,心肌细胞肌节排列更加整齐且数量丰富,更接近成熟心肌细胞形态。结论用p53 modRNA处理人诱导多能干细胞来源的早期心肌细胞,可一定程度上改善细胞成熟的相关指标。

Objective To explore the effect of p53modRNA on promoting the maturation of early cardiomyocytes derived from human pluripotent stem cells.Methods The early cardiomyocytes derived from human induced pluripotent stem cells were treated with p53 modRNA.The expression of cardiomyocyte maturation related gene mRNA was detected by qRT-PCR,the expression of cardiomyocyte maturation related protein was detected by Western blot,and the morphology and sarcomere development of cardiomyocytes were observed by cell immunofluorescence staining.Results The results of qRT-PCR showed that after p53 modRNA treatment,the expression of cardiomyocyte proliferation-related genes decreased,the level of lipid metabolism increased,and the expression of myocardial specific genes increased,suggesting that the intracellular structure of cardiomyocytes may be more mature.The results of Western blot showed that the expression of cTnI protein in the transfeetion group was higher than that in the non-transfection group.The results of immunofluorescence staining showed that compared with the non-transfection group,the morphology of cells in the transfection group tended to be long rod type,and the sarcomere of cardiomyocytes was arranged more neatly and abundantly,which was closer to the morphology of matur cardiomyocytes.Conclusion Early cardiomyocytes derived from human pluripotent stem cells treated with p53 modRNA can improve the related indexes of cell maturation to some extent.