MyoD基因诱导大鼠成纤维细胞转化为成肌细胞的研究  

Experimental studies on the differentiation of fibroblasts into myoblasts induced by MyoD genes in rats

作　　者：范慧敏[1] 李阳[2] 卢蓉[1] 汪进益[1] 刘中民[1] 

机构地区：[1]同济大学附属东方医院心脏外科,上海200120 [2]同济大学附属东方医院细胞治疗中心,上海200120

出　　处：《同济大学学报（医学版）》2007年第4期13-16,共4页Journal of Tongji University(Medical Science)

基　　金：上海市曙光计划(04SG25)

摘　　要：目的探讨MyoD在体外诱导大鼠成纤维细胞分化为成肌细胞中的作用。方法将大鼠成纤维细胞分成2组:未转染组(N组)和MyoD基因转染组(M组);利用慢病毒表达系统,将大鼠MyoD基因转入大鼠成纤维细胞中,Blasticidin筛选;用RT-PCR和Western blot检测MyoD的表达;免疫组织化学检测MyoD、desmin及-αactin的表达;电子显微镜观察转染前后细胞形态的变化。结果RT-PCR和Western blot可检测出MyoD基因转染后细胞表达MyoD mRNA及蛋白质;免疫细胞化学检测MyoD表达在基因转染的细胞中为阳性,desmin、α-actin阳性表达只出现在M组;电子显微镜观察转染后的细胞胞浆中有微丝以及微管等结构。结论MyoD基因可诱导大鼠成纤维细胞分化成的细胞具有成肌细胞特征,为进一步研究MyoD基因诱导的成肌细胞及在大鼠心梗损伤修复中的作用提供了实验基础。Objective To explore the effect of myogenic transcription factor myogenic determination （MyoD） gene on the differentiation of fibroblasts into myoblasts in rats in vitro. Methods Two groups were demonstrated： non-treated fibroblasts （N group） and MyoD gene-transfected fibroblasts （M group）. According to Lentiviral Expression System protocol, MyoD gene was transfected into fibroblasts by lipotransfection method followed by Blasticidin selection. The mRNA and protein expressions of MyoD in the transfectants were detected by RT-PCR and Western blot, respectively. Immumocytochemical methods were used to study the expressions of MyoD, desmin and α-actin. The morphological structure changes of cells before and after transfection were observed by electron microscopy. Results The mRNA and protein expressions of MyoD were detected in the MyoD-transfected fibroblasts by RT-PCR and Western blot, respectively. Moreover, the expressions of MyoD were observed by Immunocytochemical methods in the MyoD -transfected cells, and desmin and α-actin were found only in M group. The structures of microfilament and microtubule were detected in transfected cells by electron microscopy. Conclusion MyoD gene can induce rat fibroblasts differentiate into myoblast-like cells, which provides an experimental foundation for cell repair after myocardial infarction.

关 键 词：成纤维细胞 肌分化 肌肉转录调节因子 大鼠 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]