IGF-1基因转染对大鼠成肌细胞增殖和分化的影响  

Effects of IGF-1 Gene Transfection on the Proliferation and Differentiation of Myoblasts in Rat

作　　者：王晓林[1] 王庸晋[2] 荣书玲[2] 马红彪[1] 

机构地区：[1]长治医学院附属和平医院儿科,046000 [2]长治医学院附属和平医院心内科

出　　处：《长治医学院学报》2013年第6期401-405,共5页Journal of Changzhi Medical College

基　　金：山西省基础研究计划项目(2012011040-3;2013011049-4);山西高校科技研究开发项目(20101150);长治市科技计划项目(20123060)

摘　　要：目的:探讨人类胰岛素样生长因子-1(hIGF-1)基因转染对骨骼肌成肌细胞增殖和分化的作用及机制。方法:分别用pLghIGF-1SN质粒和对照质粒pLgGFPSN转染成肌细胞作为IGF组和GFP组,未转染成肌细胞作为细胞对照(Control)组。转染后用免疫细胞化学、RT-PCR检测hIGF-1基因在成肌细胞中的表达。MTT方法检测成肌细胞增殖能力,流式细胞仪检测成肌细胞的细胞周期变化。通过检测细胞融合率与磷酸肌酸激酶(CK)含量检测IGF-1基因转染对成肌细胞分化的影响。结果:基因转染后免疫细胞化学和RT-PCR检测发现IGF组细胞有hIGF-1表达,GFP组和Control组细胞未见hIGF-1表达;MTT检测显示IGF组成肌细胞光吸收值增大,与Control组和GFP组细胞相比,差异有显著性(P<0.05);流式细胞仪检测显示:与对照组和GFP组相比,IGF组G1期细胞百分比显著下降(P<0.05),S期细胞百分比则显著增加(P<0.05)。与对照组和GFP组相比,IGF组成肌细胞融合率和CK合成显著增加(P<0.05)。结论:IGF-1基因转染对成肌细胞的增殖与分化均具有促进作用,其作用是通过减少G1期成肌细胞数量,增加S期细胞数量实现的。Objective：To investigate the effects of human insulin like growth factor-1（hIGF-1）on the proliferation and differentiation of myoblasts in rats.Methods：Myoblasts were isolated from several SD rats,cultured,purified,and transfected with plasmid pLghIGF1SN and pLgGFPSN.The myoblasts were divided into IGF group（myoblasts transfected with pLghIGF-1SN,GFP group（myoblasts transfected with pLgGFPSN）,and Control group（untransfected myoblasts）.The expression of hIGF-1in myoblasts was investigated by immunocytochemistry and reverse transcription polymerase chain reaction（RT-PCR）.MTT method was used to determine the proliferation of myoblasts,and flowcytometer analysis was used to observe cell cycle changes of myoblasts.We detected the cell fusion rate and the content of creatine kinase（CK）to investigate the effects of IGF-I gene transfection on the differentiation of myoblasts.Results：After be transfected with pLghIGF-1SN,the expression of hIGF in myoblasts could be detected by immunocytochemistry and RT-PCR,but not in GFP group or control group.MTT assay showed that the absorbance value increased in IGF group compared with Control group and GFP group（P〈0.05）.Compared with control group and GFP group,the cellular proportion in S period was significantly increased and the proportion in G1period was significantly decreased after hIGF-1gene transfection（P〈0.05）.The fusion rate of myotube and quantity of CK in IGF group were significantly increased compared with Control group and GFP group（P〈0.05）.Conclusion：Gene transfection of hIGF-1can enhance the proliferation and differentiation of myoblasts via increasing the number of myoblasts at S phase and decreasing the number of myblasts at G1phase.

关 键 词：类胰岛素样生长因子-1 成肌细胞 增殖 分化 

分 类 号：R392.12[医药卫生—免疫学]