大鼠骨骼肌成肌细胞与乳鼠心肌细胞共培养对缝隙连接蛋白43表达的影响  

Effects of co-culturing with cardiomycyte on skeletal myoblasts connexin43 gene expression

作　　者：刘丽霞[1] 邵素霞[1] 张雷[1] 陈红霞[1] 赵春芳[1] 

机构地区：[1]河北医科大学组织学与胚胎学教研室,石家庄050017

出　　处：《解剖学报》2011年第4期489-493,共5页Acta Anatomica Sinica

基　　金：河北省科技厅指令计划资助项目(07276101D-129);河北医科大学2005年青年基金资助项目(0000039)

摘　　要：目的观察体外共培养的乳鼠心肌细胞与大鼠骨骼肌成肌细胞(简称L6细胞)以及乳鼠心肌细胞与转染外源性缝隙连接蛋白43(Cx43)基因的L6细胞(简称L6-Cx43细胞)间Cx43蛋白的表达情况,从而探讨共培养时心肌环境对成肌细胞的作用。方法将乳鼠心肌细胞与4',6-二脒基-2-苯基吲哚标记的大鼠骨骼肌成肌细胞进行共培养,用免疫荧光双标方法检测共培养后两种细胞中Cx43的表达定位;用Western blotting方法检测共培养对成肌细胞中Cx43蛋白表达量的影响及共培养时间对成肌细胞Cx43蛋白表达水平的影响。结果免疫荧光双标结果显示,共培养后两种细胞接触的细胞膜上均有Cx43的表达;Western blotting结果进一步证实,共培养后成肌细胞中Cx43蛋白的表达增多,L6-Cx43细胞中Cx43增多更明显。结论心肌环境能够促进成肌细胞Cx43蛋白的表达,且两种细胞接触是必需的,转染外源性Cx43可以提高这种作用。Objective To explore the effect of the cardiac environment on skeletal myoblasts by measuring connexin43 （Cx43） gene expression in L6 cells and L6-Cx43 cells when co-cultured with cardiomyocytes. Double immunofluorescence staining was applied to detect the location of Cx43 in both L6 and L6-Cx43 cells. Methods Western blotting was used to detect Cx43 gene expression on protein level at different co-cuhuring time point. Results Data from Double immunofluoreseence indicated that, Cx43 was expressed on interface between adjacent L6 cells and cardiomyocytes or between adjacent L6-Cx43 cells and cardiomyocytes when the two cell types were co-cultured in vitro. Western blotting results showed that Cx43 protein level was increased in L6 cells and even more increased in L6-Cx43 cells. Conclusion Cardiac enviroment may induce skeletal myoblasts Cx43 gene expression, and direct cell-cell contact is necessary. This effect can be enhanced by Cx43 transfection.

关 键 词：骨骼肌成肌细胞 心肌细胞 共培养 缝隙连接蛋白43 缝隙连接 免疫印迹法 大鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]