SDF-1对骨髓间充质干细胞迁移和细胞外基质形成的影响

Effects of SDF-1 on migration and extracellular matrix formation of bone marrow mesenchymal stem cells

目的探讨基质细胞衍生因子(stromal cell-derived factor-1,SDF-1)对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)迁移、胶原RNA表达及细胞膜片细胞外基质表达的影响。方法CCK-8和Transwell细胞迁移实验观察SDF-1对BMSCs增殖和迁移的影响;实时荧光定量PCR检测BMSCs胶原RNA的表达水平;蛋白质印迹法检测BMSCs细胞膜片胶原蛋白的表达水平;酶联免疫吸附试验检测BMSCs细胞膜片培养基上清透明质酸(HA)的表达水平。结果SDF-1趋化BMSCs迁移作用的强度与浓度相关,对细胞的增殖无明显影响;SDF-1降低BMSCsⅠ型胶原与Ⅲ型胶原RNA和蛋白相对表达量的比值,并显著升高BMSCs细胞膜片透明质酸表达水平(P<0.05)。结论SDF-1在适宜的浓度范围内,可显著促进BMSCs的迁移,降低Ⅰ型胶原与Ⅲ型胶原RNA和蛋白相对表达量的比值,并可提高透明质酸的表达水平。

Objective To investigate the effects of stromal cell-derived factor-1(SDF-1)on the migration,collagen RNA expression and extracellular matrix expression of rat bone marrow mesenchymal stem cells(BMSCs).Methods The effects of SDF-1 on BMSCs proliferation and migration were observed by CCK-8 assay and Transwell cell migration assay,respectively.The collagen RNA expression in BMSCs was detected by real-time fluorescence quantitative PCR.The collagen protein expression in BMSCs cell sheets was detected by Western blotting.The hyaluronic acid(HA)expression in BMSCs cell sheet culture medium was detected by enzyme-linked immunosorbent assay.Results The chemotactic effect of SDF-1 on BMSCs migration was concentration-dependent,while it had no significant effect on cell proliferation.SDF-1 decreased the ratio of RNA expression to protein expression in typeⅠcollagen and typeⅢcollagen,and significantly increased the HA expression level in BMSCs cell sheets(P<0.05).Conclusion Within a suitable concentration range,SDF-1 can significantly promote BMSCs migration,reduce the ratio of RNA expression to protein expression in typeⅠcollagen to typeⅢcollagen,and enhance HA expression levels.