基质细胞衍生因子-1与碱性成纤维细胞因子对体外骨髓间充质干细胞增殖、分化和趋化的影响

The effect of SDF-1 and bFGFon proliferation,differentiation and chemotaxis of bone marrow-derived stem cells

目的研究基质细胞衍生因子-1(SDF-1)与碱性成纤维细胞因子(bFGF)对比格犬骨髓间充质干细胞(BMSCs)增殖、分化和趋化的的影响。方法分离培养和鉴定比格犬BMSCs,利用不同浓度SDF-1与bFGF分别对比格犬BMSCs进行干预,检测不同浓度SDF-1与bFGF对BMSCs的增殖作用,检测Ⅰ型胶原、III型胶原、碱性磷酸酶(ALP)、S100钙离子结合蛋白A4(S100A4)等成骨、成纤维相关基因以及C-X-C家族趋化因子受体4(CXCR4)趋化相关基因的表达。结果SDF-1在100、200、300 ng/ml时对BMSCs均有促进作用,其中200、300 ng/ml的效果更为显著;bFGF在20、50、100 ng/ml时对BMSCs均有促进作用,其中50 ng/ml时细胞增殖最为显著。SDF-1和bFGF均可显著增强Ⅰ型胶原及S100A4的表达;bFGF增加Ⅲ型胶原及S100A4表达的作用较SDF-1更明显;bFGF可以抑制ALP的表达;SDF-1能够增加CXCR4的表达。结论SDF-1和bFGF均能促进BMSCs的增殖,并刺激BMSCs向成纤维细胞方向分化,促使抑制牙周膜矿化的基因高表达。

Objective To investigate the effect of stromal cell derived factor-1(SDF-1)and basic fibroblast growth factor(bFGF)on proliferation,differentiation and chemotaxis of bone marrow mesenchymal stem cells(BMSCs)of Beagle dog.Methods The BMSCs were cultured and identified in vitro.The cells were stimulated by using different concentrations of SDF-1 and bFGF.MTT test was used to detect the influence of SDF-1 and bFGF on the proliferation of BMSCs.Then,RT-PCR was used to detect the expressions of osteogenic and fibrogenic factors as well as chemokine related genes such as collagen I,collagen III,alkaline phosphatase(ALP),and S100 calcium binding protein A4(S100A4).Results SDF-1 promoted the proliferation of the BMSCs at concentrations of 100,200 and 300 ng/ml,and the effect is notable at concentrations of 200 and 300 ng/ml.bFGF also facilitated the proliferation of the BMSCs at concentrations of 20,50 and 100 ng/ml,and the effect was striking at 50 ng/ml.Both SDF-1 and bFGF could significantly increase the mRNA expression of collagen I and S100A4,and the effect of bFGF on S100A4 expression was more obvious than that of SDF-1.Moreover,bFGF inhibited ALP expression and SDF-1 increased the expression of CXCR4.Conclusion Both SDF-1 and bFGF can promote the proliferation of the BMSCs,stimulate the BMSCdifferentiate into fibroblasts,and promote the gene expression of inhibiting periodontal ligament mineralization.