CAJ | 학술논문

目的探讨转化生长因子β1 (TGF-β1)诱导骨骼肌肌源性干细胞(muscle-derived stem cells,MDSCs)向肌成纤维母细胞分化过程中的信号通路.方法采用差速贴壁法分离出大鼠骨骼肌肌源性干细胞,培养基中加入TGF-β1和细胞外信号调节激酶(extracellular signal-regulated kinase,Erk)特异性拮抗剂PD98059.将体外培养的细胞分为三组:对照组(A组),10 ng/ml TGF-β1组(B组),10 ng/ml TGF-β1+ 80μM PD98059组(C组).利用免疫细胞化学染色方法检测细胞表型的变化,RT-PCR、Western-blot检测细胞中CTGF和COL Ⅰ表达和合成水平.结果在体外,TGF-β1能诱导MDSCs高表达α-SMA、CTGF和COLⅠ,抑制MDSCs合成Sac-1;Erk特异性拮抗剂PD98059能抑制TGF-β1的作用,显著降低细胞CTGF和COLⅠ mRNA的表达水平,减低CTGF和COLⅠ合成,增加Sac-1的合成.结论 TGF-31能在体外诱导骨骼肌MDSCs向肌成纤维母细胞分化,TGF-β1-Erk-CTGF信号传导通路参与细胞分化过程.
목적 탐토전화생장인자β1 (TGF-β1)유도골격기기원성간세포(muscle-derived stem cells,MDSCs)향기성섬유모세포분화과정중적신호통로.방법 채용차속첩벽법분리출대서골격기기원성간세포,배양기중가입TGF-β1화세포외신호조절격매(extracellular signal-regulated kinase,Erk)특이성길항제PD98059.장체외배양적세포분위삼조:대조조(A조),10 ng/ml TGF-β1조(B조),10 ng/ml TGF-β1+ 80μM PD98059조(C조).이용면역세포화학염색방법검측세포표형적변화,RT-PCR、Western-blot검측세포중CTGF화COL Ⅰ표체화합성수평.결과 재체외,TGF-β1능유도MDSCs고표체α-SMA、CTGF화COLⅠ,억제MDSCs합성Sac-1;Erk특이성길항제PD98059능억제TGF-β1적작용,현저강저세포CTGF화COLⅠ mRNA적표체수평,감저CTGF화COLⅠ합성,증가Sac-1적합성.결론 TGF-31능재체외유도골격기MDSCs향기성섬유모세포분화,TGF-β1-Erk-CTGF신호전도통로삼여세포분화과정.
Objective To investigate the signal pathway of TGF-β1 in the process of muscle-derived stem cell (MDSCs) differentiation towards myofibroblasts.Methods MDSCs were isolated from rat skeletal muscles via differential adhesion method.TGF-β1 and PD98059,a specific antagonist to extracellular signal-regulated kinase (Erk) were added in the culture medium.Three cell culture groups were set up:group A,control; group B,10 ng/ml TGF-β1; group C,10 ng/ml TGF-β1 + 80 μM PD98059.Immunocytochemistry was employed to determine cell phenotypes.RT-PCR and Western Blot were used to detect the expression of Sca-1,α-SMA,CTGF and COL Ⅰ in isolated cells under the stimulation of TGF-β1 alone or combination of TGF-β1 and PD98059.Results In vitro,TGF-β1 upregulated the expression of α-SMA,CTGF and COL Ⅰ and inhibited the synthesis of Sca-1 in MDSCs.However,PD98059 counteracted TGF-β1,resulting in decreased mRNA expression and protein synthesis of CTGF and COL Ⅰ and increased protein synthesis of Sca-1.Conclusion TGF-β1 induces the in vitro differentiation of skeletal MDSCs towards myofibroblasts.TGF-β1-Erk-CTGF signal pathway plays an important role in the cell differentiation process.