缬沙坦诱导人脐带间充质干细胞向心肌样细胞分化

doi:10.3969/j.issn.0529-1356.2014.06.019

摘要/Abstract

摘要：

目的探讨缬沙坦诱导人脐带间充质干细胞（hUCMSCs）向心肌样细胞分化的潜能。方法 hUCMSCs经缬沙坦诱导后培养1～3 周，应用心肌肌钙蛋白T（cTnT）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(cTnI)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hUCMSCs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cTnI和心肌相关转录因子GATA4蛋白表达，对照组较少。RT-PCR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。cTnI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。

关键词: 缬沙坦, 诱导, 分化, 心肌样细胞, 免疫组织化学, 免疫荧光, 人脐带间充质干细胞

Abstract:

Objective To explore the feasibility of differentiating the human umbilical cord-derived msenchymal stem cells (hUCMSCs) to myocardiocyte-like cells after induction with valsartan. Methods The hUCMSCs were induced by valsartan and cultured for 1 week, 2 weeks and 3 weeks respectively. The antibodies against cardiac troponin T (cTnT）and recombinant GATA binding protein 4 (GATA4）were used to identify the cells which were induced to differentiate by immunocytochemistry and immunofluorescence. The mRNA expressions of cardiac troponin I(cTnI), myocardial-related transcription factor and myocardial development-related genes were detected by RT-PCR to identify the feasibility of differentiating hUCMSCs and inducing capacity of valsartan. Results The morphology of normal hUCMSCs showed uniform fibrous and whirlpool-like shape. They became significantly smaller and short spindle shape after valsartan induction. Immunochemical and immunofluorescence results showed that the expression of cTnT and GATA4 protein in the induced group was positive and seldom in the control group. RT-PCR results showed that the mRNA expression of cardiac-related transcription factor Tbx5, GATA4 and Nkx2.5 was spontaneously and continuously in the hUCMSCs. They increased after being induced by valsartan 1 week and then declined. The mRNA expression of cTnI was detected at 3 weeks, but the atrial natriuretic polypeptide（ANP）and β-myosin heavy chain (β-MHC) were not detected. Conclusion The hUCMSCs have capacity to be induced to differentiate into cardiomyocyte-like cells by valsartan.

Key words:

Valsartan, Inducation, Differentiation, Cardiomyocyte-like cell, Immunohistochemistry, Immunofluorescence, Human umbilical cord-derived msenchymal stem cell

何万里李永海杨军政林俊堂郭志坤徐振平*. 缬沙坦诱导人脐带间充质干细胞向心肌样细胞分化[J]. 解剖学报, 2014, 45(6): 841-846.

HE Wan-li LI Yong-hai YANG Jun-zheng LIN Jun-tang GUO Zhi-kun XU Zhen-ping*. Differentiation of cardiomyocyte-like cells from human umbilical cord-derived mesenchymal stem cells induced by valsartan[J]. AAS, 2014, 45(6): 841-846.

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