碱性成纤维细胞生长因子拮抗转化生长因子—β1对人α1（I）胶原基因的启动激活

目的　探讨碱性成纤维细胞生长因子 (b FGF)对人α1 ( )胶原基因启动子活性的影响 ,以及与转化生长因子 - β1 (TGF- β1 )之间的相互作用 ,为防治增生性瘢痕提供依据。　方法　正常皮肤及瘢痕成纤维细胞原代、传代培养。采用 Fu GENE转染试剂 ,分别瞬间转染含人α1 ( )胶原基因 5'端序列 - 2 .5 kb与报告基因氯霉素乙酰基转移酶(CAT)的重组体 ph COL2 .5至正常皮肤及瘢痕成纤维细胞。 ELISA法测定 b FGF及 TGF- β1 作用 2 4小时后 ,转染ph COL2 .5的两种成纤维细胞的报告基因 CAT表达量。　结果　b FGF能抑制转染 ph COL2 .5重组体的正常皮肤及瘢痕成纤维细胞 CAT表达量 ,且能拮抗 TGF-β1 对转染 ph COL2 .5重组体的两种成纤维细胞 CAT表达的上调作用。与对照组相比有统计学意义 (P<0 .0 5)。　结论　正常皮肤及瘢痕成纤维细胞中 ,b FGF均能抑制人 α1 ( )胶原基因的启动转录 ,且能拮抗 TGF-β1 对人α1 ( )胶原基因启动活性的上调作用 ,b FGF抗纤维机制有望为增生性瘢痕的防治提供新思路

Basic fibroblast growth factor inhibits promoter activities of human alpha 1(I) procollagen gene induced by transforming growth factor-beta 1]

OBJECTIVE: To investigate the effects of basic fibroblast growth factor (bFGF) on the promoter activities of human alpha 1(I) procollagen gene and the interaction between bFGF and transforming growth factor-beta 1 (TGF-beta 1). METHODS: Fibroblasts of the hypertrophic scar and normal skin from a 3-year-old patient were primarily cultured and subcultured in vitro. Both of the fibroblasts were transient transfected with phCOL 2.5, containing -2.5 kb of 5'f lank sequence of human alpha 1(I) procollagen gene and CAT reporter gene by FuGENE transfection reagent; and treated thereafter by 16 ng/ml bFGF, 2 ng/ml TGF-beta 1 and 16 ng/ml bFGF + 2 ng/ml TGF beta 1 for 24 hours. The relative CAT expression values were determined by CAT-ELISA. RESULTS: TGF-beta 1 strongly induced the CAT expression level, however, bFGF not only inhibited the basal CAT expression but also reduced the CAT expression up-regulated by TGF-beta 1 in normal skin and hypertrophic scar fibroblasts (P < 0.05). CONCLUSION: bFGF can reduce the promoter activities of human alpha 1(I) procollagen gene and antagonize the role of TGF-beta 1 in up-regulating the promoter activities of human alpha 1(I) procollagen gene in normal skin and hyertrophic scar fibroblasts.