Myocardin重组蛋白表达及其在人骨髓间充质干细胞重编程为血管平滑肌细胞中的作用  

Expression of recombinant myocardin and its effect on human bone marrow mesenchymal stem cell reprogramming into vascular smooth muscle cells

作　　者：麦丽萍 杨翔宇 吴岳恒 余细勇 何国东 李晓红 MAI Li-ping;YANG Xiang-yu;WU Yue-heng;YU Xi-yong;HE Guo-dong;LI Xiao-hong(Medical Research Center of Guangdong General Hospital,Guangdong Academy of Medical Sciences,Key Laboratory of Structural Heart Disease in Guangdong Southern China,Guangzhou 510080,China;College of Pharmacy,Guangzhou Medical University,Guangzhou 511436,China)

机构地区：[1]广东省人民医院,广东省医学科学院医学研究部,广东省华南结构性心脏病重点实验室,广东广州510080 [2]广州医科大学药学院,广东广州511436

出　　处：《中国病理生理杂志》2018年第8期1409-1414,共6页Chinese Journal of Pathophysiology

基　　金：广东省科技计划(No.2017A070701013;No.2017B090904034); 广州市科技计划(No.201704020126)

摘　　要：目的:研究心肌素(myocardin)重组蛋白的表达及分离纯化方法,并探讨myocardin重组蛋白对人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,h BMSCs)向血管平滑肌细胞转分化的作用。方法:构建myocardin原核表达载体,摸索最佳诱导myocardin重组蛋白表达的方法及优化分离纯化方式。使用纳米转导试剂包裹myocardin形成纳米-蛋白复合体,转染h BMSCs,观察蛋白转导效率;蛋白转导成功后,检测血管平滑肌的特异性标志物平滑肌肌球蛋白重链的表达情况,观察h BMSCs向平滑肌细胞转化情况。结果:与传统方法相比较,高浓度法诱导myocardin重组蛋白的表达量更高,新采用的洗脱方式能获取更多的myocardin重组蛋白;纳米-myocardin复合体可成功转染h BMSCs并诱导其向血管平滑肌样细胞转化。结论:Myocardin重组蛋白能被高效转导到细胞内,并促使h BMSCs重编程为血管平滑肌样细胞。AIM： To optimize the expression and purification of recombinant myocardin,and to explore the effect of recombinant myocardin on the transdifferentiation of human bone marrow mesenchymal stem cells（ h BMSCs） into vascular smooth muscle cells（ v SMCs）. METHODS： The prokaryotic expression vector for myocardin was constructed.The best ways to induce the expression of myocardin recombinant protein and to optimize the isolation and purification methods were explored. Nano-transducers were applied to encapsulate myocardin to form nano-protein complexes. Nanomyocardin was transducted into h BMSCs and the protein transduction efficiency was measured. After transduction of the protein was successful,smooth muscle myosin heavy chain,a specific marker of v SMCs,was detected,and the transformation of h BMSCs into v SMCs was also observed. RESULTS： Compared with the traditional methods,the expression of myocardin recombinant protein was higher by high-concentration induction method,and the new elution method was able to obtain more myocardin recombinant protein. The nano-myocardin complexes were successfully transduced into the h BMSCs and induced the transformation of h BMSCs into v SMCs. CONCLUSION： The recombinant myocardin was efficiently transduced into the cells and reprogrammed h BMSCs into v SMCs.

关 键 词：心肌素 重组蛋白 平滑肌肌球蛋白重链 骨髓间充质干细胞 细胞重编程 

分 类 号：R318.11[医药卫生—生物医学工程] R363[医药卫生—基础医学]