背景：在皮肤修复及瘢痕形成过程中如何控制胶原蛋白的有序形成是值得关注的热点。目的：探讨碱性成纤维细胞生长因子与胰岛素样生长因子1对体外培养大鼠骨髓间充质干细胞增殖及胶原合成的影响。方法：体外分离培养大鼠骨髓间充质干细胞，进行成脂及成骨分化鉴定，在第3代细胞培养液中分别添加碱性成纤维细胞生长因子、胰岛素样生长因子1以及两者联合干预，以加入正常培养基为对照组。培养12，24，48，72，96 h时采用MTT检测细胞增殖情况，培养10 d时采用RT-PCR检测Ⅰ型胶原、Ⅲ型胶原mRNA表达，Western Blot检测Ⅰ型胶原、Ⅲ型胶原蛋白表达。结果与结论：①与对照组比较，单独应用碱性成纤维细胞生长因子可显著促进骨髓间充质干细胞增殖，抑制Ⅰ型胶原、Ⅲ型胶原表达。②单独应用胰岛素样生长因子1可显著促进骨髓间充质干细胞增殖，促进Ⅰ型胶原、Ⅲ型胶原表达。③两种生长因子联用相比单独应用，促进增殖效果更为显著，同时使Ⅰ型胶原、Ⅲ型胶原表达趋于正常水平。④结果表明碱性成纤维细胞生长因子、胰岛素样生长因子1联合应用在促进骨髓间充质干细胞增殖的同时能够对Ⅰ型胶原、Ⅲ型胶原的表达有所限制，可能有助于控制瘢痕形成及促进创伤愈合。%BACKGROUND:How to control the orderly formation of colage in skin repair and scarring process is worthy of attention. OBJECTIVE: To investigate the effect of basic fibroblast growth factor (bFGF) combined with insulin-like growth factor 1 (IGF-1) on the proliferation and colagen synthesis of rat bone marrow mesenchymal stem celsin vitro. METHODS:Rat bone marrow mesenchymal stem cels were isolated and cultured to induce adipogenic differentiation assessed by oil red O staining and osteogenic differentiation identified by alizarin red stainingin vitro. Passage 3 cels were cultured in the medium containing bFGF, IGF-1, combination of them or the control fluid, respectively. MTT assay was used to detect cel proliferation at 12, 24, 48, 72 and 96 hours of culture. The expression of type I colagen and type III colagen were detected by RT-PCR and western blot after 10 days of incubation. RESULTS AND CONCLUSION:Compared with the control group, bFGF or IGF-1 alone significantly promoted the proliferation of bone marrow mesenchymal stem cels, and inhibited the expression of type I colagen and type III colagen. After combined use of bFGF and IGF-1, the proliferation of bone marrow mesenchymal stem cels was improved more significantly, and the expression of type I colagen and type III colagen returned to normal levels. These findings indicate that the combination of IGF-1 and bFGF can promote proliferation of bone marrow mesenchymal stem cels and restrain the expression of type I colagen and type III colagen, which may be helpful for control and repair of scar formation during wound healing.
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