摘要
目的通过检测移植经干扰乙酰化作用基因Gcn5表达重组质粒ZJ3转染间充质干细胞（mesenchymal stem cells，MSCs）的Wistar大鼠心肌组织内乙酰化酶活性及乙酰化作用基因Gcn5、心肌发育基因GATA4的表达量，探讨体内心肌微环境下乙酰化修饰在干细胞特化心肌样细胞转录调控中的作用。方法提取干扰组蛋白乙酰化酶Gcn5表达重组质粒ZJ3，经脂质体包载转染MSCs24h后移植至Wistar大鼠心肌组织内，2w后检测移植区域心肌组织内乙酰化酶活性及乙酰化作用基因Gcn5、心肌发育基因GATA4的表达量。结果实验组心肌组织乙酰化酶活性较正常对照组、阴性对照组及试剂对照组均有显著性降低；实验组心肌组织内Gcn5及GATA4表达量较正常对照组、阴性对照组及试剂对照组均有明显减弱。结论相关特异性基因的检测结果显示封阻干细胞染色质组蛋白乙酰化修饰后，在体内心肌微环境诱导下亦可以抑制干细胞特化心肌细胞转录过程，这为进一步研究组蛋白末端乙酰化修饰与干细胞分化调节机制奠定了实验室基础。
Objective To explore the effects of histone acetylation modification on the regulation of mesenchymal stem cells （MSCs） differentiation into cardiomyocytes in myocardium microenvironment, by detecting the acetylase activity, expression of histone acetylation gene Gcn5 and myocardial gene GATA4 in myocardium transplanted by MSCs transfected with plasmid ZJ3. Methods Extracted the plasmid containing short hairpin RNA （shRNA） against Gcn5 gene （ZJ3） and transfected it into MSCs. After 24 hours, the MSCs were transplanted into the rat myocardium tissues and the acetylase activity, the expression of acetylation gene and myocardium development gene were detected after two weeks. Results The acetylase activity in the experimental group was significantly lower than all control groups; Gcn5 and GATA4 expression in myocardium of experimental group had significant reduction compared with the control groups. Conclusion Inhibition of histone acetylation in the MSC cells can inhibit the transcription process of specialized myocardium in the myocardium microenvironment. The result establishes foundation for researches of histone acetylation and mechanisms of MSCs differentiation.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2008年第2期159-163,共5页
Chinese Journal of Medical Genetics
基金
国家自然科学基金（30471837）
