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目的 探讨地塞米松与转化生长因子 β1 (TGFβ1 )之间的相互作用及对人α1 (Ⅰ )前胶原基因启动转录的影响。方法 人皮肤及瘢痕成纤维细胞原代、传代培养。采用FuGENE转染试剂 ,分别瞬间转染含人α1 (Ⅰ )胶原基因 5′侧翼序列 - 2 5kb与报告基因氯霉素乙酰基转移酶 (CAT)的重组体 phCOL2 5至人皮肤及瘢痕成纤维细胞。ELISA法测定地塞米松及TGFβ1 作用 2 4h后 ,转染了phCOL2 5的 2种成纤维细胞的报告基因CAT表达量。 结果 地塞米松能抑制转染了 phCOL2 5重组体的人皮肤及瘢痕成纤维细胞CAT表达量 ,且能拮抗TGFβ1 对转染了 phCOL2 5重组体的 2种成纤维细胞CAT表达的上调作用 (P <0 0 5)。结论 在正常皮肤及瘢痕成纤维细胞中 ,地塞米松均能抑制人α1 (Ⅰ )前胶原基因的启动转录 ,且能拮抗TGFβ1 对人α1 (Ⅰ )前胶原基因的转录激活
Objective To investigate the interaction between dexamethasone and transforming growth factor β1 (TGFβ1) and its effect on the transcription of human α1 (Ⅰ) procollagen gene. Methods Human skin and scar fibroblasts primary, subculture. FuGENE transfection reagent was used to transiently transfect human skin and scar fibroblasts containing 5 ’flanking sequence of human α1 (Ⅰ) collagen gene and 25 kD of recombinant reporter gene chloramphenicol acetyltransferase (CAT) cell. ELISA method was used to determine the expression level of CAT in two fibroblasts transfected with phCOL2 5 after treated with dexamethasone and TGFβ1 for 24 h. Results Dexamethasone inhibited CAT expression in human skin and scar fibroblasts transfected with phCOL2 5 recombinant and inhibited the up-regulation of CAT expression in two fibroblasts transfected with phCOL2 5 recombinant (P <0 0 5). Conclusion Dexamethasone can inhibit the transcriptional activation of human α1 (Ⅰ) procollagen gene and inhibit the transcriptional activation of human α1 (Ⅰ) procollagen gene in normal skin and scar fibroblasts