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目的 :探讨胰岛素样生长因子 1(IGF 1)和胰岛素对人α1(Ⅰ )前胶原 (COL1A1)基因启动子的调控作用及肿瘤坏死因子α(TNFα)、干扰素γ(IFNγ)、干扰素α(IFNα)对这一作用的影响。方法 :构建含COL1A1基因启动子序列与氯霉素乙酰基转移酶 (CAT)报告基因的重组体pCOLH0 2 7与pCOLH2 5 ,它们分别含该启动子序列 2 68～ + 4 2bp( 0 2 7kb)与 2 4 83～ + 4 2bp( 2 5kb)片段。将这 2个重组体瞬时转染人皮肤成纤维细胞 ,加入IGF 1或胰岛素 ,部分细胞还加入TNFα或IFNγ或IFNα,测定CAT活性。结果 :13nmol/LIGF 1使转染后的这 2个重组体活性分别增高 3 68倍与 4 0 4倍。 2 5 μmol/L的胰岛素亦使之分别增高 3 69倍与 3 93倍。TNFα、IFNγ、IFNα能降低IGF 1与胰岛素诱导的pCOLH2 5活性。结论 :IGF 1和胰岛素能在转录水平促进COL1A1基因的表达 ,TNF α、IFNγ、IFNα能抑制IGF 1和胰岛素的这一作用。
OBJECTIVE: To investigate the regulatory effect of insulin-like growth factor 1 (IGF 1) and insulin on the promoter of human α1 (COL1A1) gene and on the expressions of tumor necrosis factor α (TNFα), interferon α (IFNα) on this effect. Methods: The recombinant pCOLH027 and pCOLH2 5 containing the promoter sequence of COL1A1 gene and the reporter gene of chloramphenicol acetyltransferase (CAT) were constructed, which contained the promoter sequence of 68 ~ +4 2bp (0 27kb) With 2 4 83 ~ + 4 2bp (25kb) fragments. The two recombinants were transiently transfected into human skin fibroblasts, IGF1 or insulin was added, and some cells were also added with TNFα or IFNγ or IFNα to measure CAT activity. Results: The activity of these two recombinants after transfection was increased by 3 68-fold and 4 0 4-fold respectively by 13 nmol / L IGF-1. 2 5 μmol / L insulin also increased by 3 69 times and 3 93 times. TNFα, IFNγ, IFNα reduced IGF1 and insulin-induced pCOLH2 5 activity. CONCLUSION: IGF1 and insulin can promote the expression of COL1A1 gene at the transcriptional level. TNFα, IFNγ and IFNα can inhibit IGF1 and insulin.