tgf-1促进骨骼肌肌源性干细胞合成胶原纤维的实验研究-experimental study on tgf - 1 promoting collagen fiber synthesis from skeletal muscle myogenic stem cells.docx

摘 要 TGF-ß1 促进骨骼肌肌源性干细胞合成胶原纤维的实验研究 目的：探讨 TGF-ß1 诱导骨骼肌肌源性干细胞(ＭＤＳＣ)合成细胞外基质的机制。 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-ß1。 将体外培养的细胞分为三组，A：对照组；B：10ng/ml TGF-ß1；C：10ng/ml TGF-ß1+10ng/ml 抗 CTGF 中和抗体。在相差显微镜下观察细胞生长情况，利用免 疫组织化学方法, RT-PCR, West-blot 检测细胞表达 CTGF、ColⅠ、ColIII 的变化 情况。 结果：在体外,TGF-ß1 能诱导 MDSC 高表达 CTGF，促进细胞合成和分泌 ColⅠ和 ColIII，CTGF 抗体能降低细胞表达和合成 ColⅠ和 ColIII，部分逆转 TGF-ß1 对 MDSC 合成 ECM 的作用。 结论：TGF-ß1 能在体外诱导 MDSC 合成和分泌细胞外基质，促进组织纤维化； TGF-ß1- CTGF 信号传导通路是介导 TGF-ß1 促进细胞外基质的合成分泌的机制之 一。 【关键词】 TGF-ß1；CTGF；肌源性干细胞;细胞外基质  Division of Hand surgery,Union Hospital of Tongji Medical College, Huazhong University of Science and Technology Master candidate: pengyunlong Supervisor: Prof.Chen Zhenbing TGF-ß1 promotes the synthesis of collagen fibers in skeletal muscle-derived stem cells Abstract Purpose Investigate the mechanism of TGF-ß1 induced synthesis of extracellular matrix in skeletal muscle-derived stem cells (MDSC). Method To isolate rat skeletal muscle-derived stem cells by differential adhesion method, medium supplemented with TGF-β1. Cultured cells were divided into three groups, A: control group; B: 10ng/ml TGF-β1; C:10ng/ml TGF-β1+ 10ng/ml CTGF antibody. Cells growth condition was observed under phase contrast microscope. The expression of CTGF, Col I and Col III was examined by RT-PCR, Western-blot and immunofluorescence staining. Results In vitro, TGF-β1 can induce MDSC to express CTGF, promote the synthesis and secretion of Col I and Col III. CTGF antibody can reduce the expression and synthesis of Col I and Col III, partially reverse the effect that TGF-β1 have on the MDSC. Conclusion TGF-β1 can induce the synthesis and secretion of extracellular matrix, thus promoting tissue fibrosis in vitro; TGF-β1-CTGF signaling pathway is one of the mechanisms that mediating TGF-β1 to stengthen the synthesis and secretion of extracellular matrix. Key Words: TGF-β1；CTGF；muscle-derived stem cells；ECM  英文缩略词表 Abbreviations 英文缩写 英文全称 中文全称 CTGF Connective tissue growth factor 结缔组织生长因