外源性rhCNTF抑制成人成肌细胞体外分化作用及其信号机制  

Effect of Exogenous rhCNTF on Myogenic Differentiation of Adult Human Myoblast in vitro and Signal Mechanism

作　　者：葛昕[1] 李亚明[1] 窦忠英[1] 

机构地区：[1]西北农林科技大学国家干细胞工程技术研究中心陕西分中心,陕西杨凌712100

出　　处：《西北农业学报》2010年第9期1-6,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(30393130;90608028);国家"973"项目(2006CB504100)

摘　　要：为探讨成肌细胞体外分化的机理,采用免疫组织化学和Western blot等方法,分析重组人睫状神经营养因子(rhCNTF)对成人成肌细胞体外分化的作用及其信号机制。结果表明:①rhCNTF可逆性抑制成人成肌细胞体外融合(P<0.01);②rhCNTF显著增加成人成肌细胞分化期间的p130阳性"储备细胞"数量(P<0.01);③Western blot分析显示,rhCNTF可诱导ERK1/2磷酸化激活、成肌分化特异标志myogenin和p21蛋白表达的抑制以及"储备细胞"特异性标志p130蛋白水平的增加。首次发现,外源性rhCNTF可通过ERK1/2信号通路的激活增加"储备细胞"数量,可逆性抑制成人成肌细胞的体外分化。为进一步开展rhC-NTF在成人骨骼肌损伤修复中的作用研究奠定了基础。To determine the role of myoblast differentiation in vitro,immunocytochemistry and Western blot were employed to observe the effects of rhCNTF（recombinant human ciliary neurotrophic factor） on myoblast differentiation of adult human in vitro.Results indicated that rhCNTF（10 ng/mL）significantly inhibited myoblast differentiation（P0.01）reversibly,and inducted multiplication of reserve cell（P0.01）.Moreover,PD098059,a specific inhibitor of ERK1/2-MAPK pathway,was able to abolish the effect of CNTF on myoblast and reserve cell.Western blot analysis also indicated that,compared with the control group,the expression of p21 and myogenin,marker of myoblast differentiation phase,was significantly increased（P0.01）,while p130,a marker of reserve cell phase was significantly reduced（P0.01）,and ERK1/2-MAPK phosphorylation was reduced（P0.01）.These results demonstrate that exogenous rhCNTF can reversibly inhibit differentiation and induce increasing of reserve cell via MAPK/ERK.These findings may have important implications for degenerative disease treatment of the adult human.

关 键 词：重组人睫状神经营养因子(rhCNTF) ERK1/2-MAPK 成肌细胞 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]