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目的 观察bFGF对人皮肤成纤维细胞增殖的影响及调控α1(I)原胶原基因序列的作用。 方法 用组织块法培养人皮肤成纤维细胞并传代，采用BrdU掺入的ELISA法，测定不同浓度的bFGF对成纤维细胞增殖的影响。构建三种人α1(I)原胶原基因5’ 侧翼序列与报告基因氯霉素乙酰基转移酶(CAT)的重组质粒，用FuGENE转染试剂转染成纤维细胞，同时用p-sv-b-GAL表达质粒转染细胞作为阳性对照组，ELISA法测定经bFGF处理后成纤维细胞CAT的表达量。 结果 在体积分数2%或10%小牛血清培养条件下，bFGF加入浓度从0.25 ng/ml增至64.00 ng/ml，作用24 h后，各组细胞增殖数值之间差异有显著性意义（P<0.05）。用三种重组质粒分别转染细胞，bFGF处理24 h后，CAT相对表达量测定结果提示， bFGF组与对照组相比差异有显著性意义（P<0.05）。 结论 bFGF对人皮肤成纤维细胞增殖有抑制作用,对胶原基因启动序列具有负性调控作用，且存在剂量依赖关系。
Objective To observe the effect of bFGF on the proliferation of human dermal fibroblasts and the effect of regulating α1 (I) procollagen gene sequence. Methods Human dermal fibroblasts were cultured and passaged by tissue block method. The effect of different concentrations of bFGF on the proliferation of fibroblasts was determined by BrdU incorporation ELISA. The recombinant plasmids containing 5 ’flanking sequences of human α1 (I) procollagen gene and chloramphenicol acetyltransferase (CAT) gene were constructed and transfected into fibroblasts with FuGENE transfection reagent. Meanwhile, p-sv-b -GAL expression plasmid transfected cells as a positive control group, ELISA assay bFGF treated fibroblasts CAT expression. Results The proliferation of bFGF was increased from 0.25 ng / ml to 64.00 ng / ml under the condition of 2% or 10% fetal bovine serum. The difference of cell proliferation between groups was significant P <0.05). After transfection of cells with three recombinant plasmids respectively, the relative expression of CAT in bFGF treated group for 24 h showed that there was significant difference between bFGF group and control group (P <0.05). Conclusion bFGF can inhibit the proliferation of human dermal fibroblasts and has a negative regulatory effect on the promoter sequence of collagen gene in a dose-dependent manner.