碱性成纤维细胞生长因子拮抗转化生长因子-β_1对人α1(I)胶原基因的启动激活

BASIC FIBROBLAST GROWTH FACTOR INHIBITS PROMOTER ACTIVETIES OF HUMAN α1(I) PROCOLLAGEN GENE INDUCED BY TRANSFORMING GROWTH FACTOR-β_1

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

Objective To investigate the effects of basic fibroblast growth factor(bFGF) on the promoter activities of human α1(I) procollagen gene and the interaction between bFGF and transforming growth factor β 1(TGF β 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 phCOL2.5, containing 2.5kb of 5′f lank sequence of human α1(I) procollagen gene and CAT reporter gene by FuGENE transfection reagent; and treated thereafter by 16ng/ml bFGF, 2 ng/ml TGF β 1 and 16ng/ml bFGF+2 ng/ml TGFβ 1 for 24 hours. The relative CAT expression values were determined by CAT ELISA. Results TGF β 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 β 1 in normal skin and hypertrophic scar fibroblasts ( P< 0.05). Conclusion bFGF can reduce the promoter activities of human αl(I) procollagen gene and antagonize the role of TGF β 1 in up regulating the promoter activities of human α1(I) procollagen gene in normal skin and hypertrophic scar fibroblasts.