ShRNA on histone acetylation induces unusual cardiac-specific protein expression in MSCs differentiation into cardiomyocytes

朱静; 冯川; 李莉; 邓兵; 田杰; 张晓萍

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ShRNA on histone acetylation induces unusual cardiac-specific protein expression in MSCs differentiation into cardiomyocytes

更新时间：2026-01-09

- ShRNA on histone acetylation induces unusual cardiac-specific protein expression in MSCs differentiation into cardiomyocytes
- Journal of Army Medical University Vol. 30, Issue 4, Pages: 303-306(2008)
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  CLC： R329.2;R341;R394.3
- Published：2008
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朱静,冯川,李莉,邓兵,田杰,张晓萍.干扰组蛋白乙酰化引发间充质干细胞心肌定向分化特定蛋白的异常表达[J].陆军军医大学学报(原第三军医大学学报),2008,30(04):303-306.

　ZHU Jing,FENG Chuan,LI Li,DENG Bing,TIAN Jie,ZHANG Xiao-ping.ShRNA on histone acetylation induces unusual cardiac-specific protein expression in MSCs differentiation into cardiomyocytes[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(04):303-306.
朱静,冯川,李莉,邓兵,田杰,张晓萍.干扰组蛋白乙酰化引发间充质干细胞心肌定向分化特定蛋白的异常表达[J].陆军军医大学学报(原第三军医大学学报),2008,30(04):303-306. DOI：

　ZHU Jing,FENG Chuan,LI Li,DENG Bing,TIAN Jie,ZHANG Xiao-ping.ShRNA on histone acetylation induces unusual cardiac-specific protein expression in MSCs differentiation into cardiomyocytes[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(04):303-306. DOI：

摘要

目的利用组蛋白乙酰化修饰关键因子Gcn5 shRNA重组质粒转染经5-氮杂胞苷(5-azacytidine

5-aza)诱导后的间充质干细胞（mesenchymal stem cells，MSCs）并检测心肌早期发育相关结构和功能蛋白表达情况，探讨组蛋白乙酰化修饰在MSCs定向分化过程中的调控作用。方法 ①以组蛋白乙酰化修饰关键因子Gcn5为靶点，用带绿色荧光蛋白（green fluorescence protein，GFP）基因的pGenSil-1作为载体，构建Gcn5 shRNA质粒。②MSCs体外扩增，5-aza诱导后，不同的时间点用脂质体包载Gcn5 shRNA共转染大鼠MSCs，24 h后相差显微镜下观察细胞生长状况，电镜下观察染色质的形态变化，采用Western blot法分别检测Gcn5、心肌结构重链蛋白(myosin heavy chain，MHC)和连接蛋白（Connexin 43

Cx43）的表达状况。结果电镜示经通用阴性质粒HK转染的MSCs细胞，形态与正常MSCs无明显差别，染色质仍以边集状态为主；经干扰质粒转染的MSCs细胞在形态上与正常MSCs细胞无明显差别，但其染色质出现明显的浓缩紧密现象。以β-actin为内参，比较干扰组和阴性对照组Gcn5、Cx43、MHC蛋白，计算出沉默效率分别为86%、58%和67%。结论转染Gcn5 shRNA质粒可干扰组蛋白乙酰化平衡，进而影响MSCs体外分化中心肌发育特定蛋白的表达。

Abstract

Objective To construct recombinant plasmid that can inhibit acetyltransferase Gcn5 expression through RNA interference

then transfect it into MSCs induced by 5-azacytidine (5-aza)

and analyze the role of acetylases in the process of MSCs differentiation. Methods MSCs were isolated from bilateral thigh bones and tibias of Wistar rats

then treated with 5-aza. Gcn5 shRNA recombinant plasmid vector was transfected into MSCs via lipofectin. The expressions of Gcn5

myosin heavy chain (MHC) and Connexin 43 (Cx43) were investigated by Western blot. Results Under electron microscope

MSCs transfected with Gcn5 shRNA recombinant plasmid displayed obvious heterochromatin

but MSCs and MSCs transfected HK plasmid showed euchromatin. The expressions of Gcn5

Cx43 and MHC decreased significantly after Gcn5 shRNA recombinant plasmid transfection. Taking β-actin as inner criterion

the silent ratio of Gcn5

MHC and Cx43 were 86%

58% and 67%

respectively. Conclusion The Gcn5 shRNA plasmids we constructed block acetylation during the process of MSCs differentiation into cardiomyocytes. Acetylation plays a very important role in the regulation of MSCs differentiation.

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