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目的探讨缬沙坦诱导人脐带间充质干细胞(h UCMSCs)向心肌样细胞分化的潜能。方法 h UCMSCs经缬沙坦诱导后培养1~3周,应用心肌肌钙蛋白T(c Tn T)和GATA结合蛋白4重组蛋白(GATA4)抗体进行免疫细胞化学和免疫荧光染色,鉴定分化后的细胞。通过RT-PCR检测心肌肌钙蛋白I(c Tn I)基因、心肌相关转录因子和心肌细胞发育相关基因的mRNA表达,进行h UCMSCs的心肌分化潜能和缬沙坦诱导能力的分析。结果人脐带间充质干细胞形态为均一的纤维状细胞,呈漩涡状排列。缬沙坦诱导后明显变小呈短梭形且不规则排列。免疫细胞化学和免疫荧光结果显示,诱导组细胞有心肌特异蛋白c Tn I和心肌相关转录因子GATA4蛋白表达,对照组较少。RTPCR结果显示,人脐带间充质干细胞自发的、持续性表达心肌相关转录因子Tbx5、GATA4和Nkx2.5的mRNA,经缬沙坦诱导1周时其mRNA表达水平略有上升,随后逐渐下降。c Tn I的mRNA在诱导第3周时出现,未见心肌发育相关基因心房利钠肽(ANP)和β-心肌肌球蛋白重链(β-MHC)的mRNA表达。结论人脐带间充质干细胞具有心肌分化潜能但无法自发分化;缬沙坦具有诱导人脐带间充质干细胞向心肌样细胞分化的能力。
Objective To investigate the potential of valsartan-induced differentiation of human umbilical cord mesenchymal stem cells (h UCMSCs) into cardiomyocyte-like cells. Methods h UCMSCs were cultured for 1-3 weeks after induction with valsartan. Immunocytochemistry and immunofluorescence staining were performed using cardiac troponin T (cTn T) and GATA4 recombinant protein (GATA4) cell. The mRNA expression of cardiac troponin I (cTn I) gene, cardiac-related transcription factor and cardiomyocyte development-related genes was detected by RT-PCR. The differentiation potential of h UCMSCs and the induction of valsartan were analyzed. Results The morphology of human umbilical cord mesenchymal stem cells was homogeneous fibrous cells, arranged in a swirling pattern. Valsartan significantly smaller after induction was short-fusiform and irregular arrangement. The results of immunocytochemistry and immunofluorescence showed that the expressions of cTn I and GATA4 in the induced group were less than those in the control group. RTPCR results showed that human umbilical cord mesenchymal stem cells spontaneously and persistently expressed mRNAs of cardiac-related transcription factors Tbx5, GATA4 and Nkx2.5, and their expression levels slightly increased at 1 week after valsartan induction, and then decreased gradually. mRNA of cTnI appeared at the third week of induction, and no mRNA expression of atrial natriuretic peptide (ANP) and beta-cardiac myosin heavy chain (beta-MHC) was found. Conclusion Human umbilical cord mesenchymal stem cells have the potential of myocardial differentiation but not spontaneous differentiation. Valsartan can induce the differentiation of human umbilical cord mesenchymal stem cells into cardiomyocyte-like cells.