利用bFGF提高直接重编程诱导成纤维细胞向心肌细胞转化效率的实验研究

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摘要目的研究碱性成纤维细胞生长因子（bFGF）对直接重编程细胞向心肌细胞转化的影响。方法取大鼠尾成纤维细胞，利用GATA-4、Mef2c、Tbx5三个基因同时转染进行直接重编程。重编程后将细胞随机分为两组，一组正常培养，另一组加入50 μg/L bFGF进行培养。干预2周后，通过检测心肌细胞特异性蛋白如肌钙蛋白T（cTNT）、α-肌球蛋白重链（α-MHC）、GATA结合蛋白4抗体（GATA-4）和连接蛋白43（CX43）的表达及分析细胞电生理特点鉴定细胞分化，并比较两组分化比率。结果 Western blot和免疫荧光检测结果显示，bFGF明显提高直接重编程细胞内cTNT、α-MHC、GATA-4和CX43的表达（P ＜ 0.01）。电生理分析显示这些细胞与心肌细胞的电生理特点极为相似，进一步证实了心肌细胞转化。结论成纤维细胞可以通过直接重编程转化为心肌细胞，bFGF可促进这一过程，这可能成为一种新的获取心肌细胞的方法。

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	任杰赵龙▲ 宋振祥李伟

关键词 ：直接重编程, 心肌细胞, 碱性成纤维细胞生长因子

Abstract：Objective To investigate the effects of basic fibroblast growth factor (bFGF) on the transformation of direct reprogramming-induced transdifferentiation of fibroblasts into cardiomyocytes. Methods Direct reprogramming was performed to induce rat tail fibroblasts into cardiomyocytes by defined factors GATA-4, Mef2c and Tbx5. Then the cells were divided into two groups, one group was cultured normally, and the other group was cultured with 50 μg/L bFGF. Two weeks later, the differentiation of cardiomyocytes was identified by detection of the expression of cardiac specific protein such as cardiac troponin T (cTNT), α-myosin heavy chain (α-MHC), GATA binding protein 4 antibody (GATA-4) and connexin 43 (CX43) and analysis of electrophysiology. And the differentiation rates of the two groups were compared. Results Western blot and immunofluorescence test showed that the expression of cTNT, α-MHC, GATA-4 and CX43 significantly up-regulated after exposure to bFGF (P ＜ 0.01). And the further analysis of electrophysiology revealed that the electrophysiological characteristics of these cells were very similar to those of cardiomyocytes, which was identified as cardiac differentiation. Conclusion Fibroblasts can be induced by direct reprogramming to differentiate into cardiomyocytes, which can be promoted by bFGF. This study may provide a new method to harvest cardiomyocytes.

Key words： Direct reprogramming Cardiomyocytes Basic fibroblast growth factor

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引用本文:

任杰赵龙▲ 宋振祥李伟. 利用bFGF提高直接重编程诱导成纤维细胞向心肌细胞转化效率的实验研究[J]. 中国医药导报, 2019, 16(2): 17-19,28.
REN Jie ZHAO Long▲ SONG Zhenxiang LI Wei. Experimental study on the efficiency of bFGF promoting direct reprogramming-induced transdifferentiation of fibroblasts into cardiomyocytes. 中国医药导报, 2019, 16(2): 17-19,28.

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