转染Sp1基因对人增生性瘢痕成纤维细胞胶原表达的影响  

Effect of recombinant Spl gene on the collagen expression of hypertrophic scar fibroblasts in vitro

作　　者：杨松林[1] 万伟东[2] 郑江红[1] 张世新[1] 高云[1] 邓辰亮[1] 

机构地区：[1]上海交通大学附属第六人民医院,200233 [2]东南大学附属中大医院

出　　处：《中华整形外科杂志》2009年第1期46-49,共4页Chinese Journal of Plastic Surgery

基　　金：国家自然科学基金资助课题（30571929）

摘　　要：目的探讨重组Sp1基因转染人增生性瘢痕成纤维细胞的可行性，以及转染Sp1基因对细胞增殖速率和Ⅰ、Ⅲ型胶原合成的影响。方法体外重组Sp1基因，借助真核表达载体系统，转入体外培养的人增生性瘢痕成纤维细胞，通过激光共聚焦显微镜观察报告基因表达，并确定细胞转染效率。应用Real-timePCR法比较转染前后Sp1 mRNA及Ⅰ、Ⅲ型胶原mRNA的表达；采用CCK8比色法，比较转染细胞与未转染细胞增殖状况。结果基因转染后，约30％的被转染细胞表达绿色荧光；Sp1 mRNA在转染组、转染空载体组、未转染组细胞中的表达分别为：5．26±0．76、1．08±0．18、1．09±0．15，转染组Sp1 mRNA相对表达量较未转染组和转染空载体组明显增高（P〈0．01，n=5）；Ⅰ、Ⅲ型胶原mRNA在转染组、转染空载体组、未转染组细胞中的表达分别为：2．49±0．40、1．88±0．30，0．96±0．18、0．95±0．18，0．97±0．15、0．93±0．13，转染组Ⅰ、Ⅲ型胶原mRNA相对表达量较未转染组和转染空载体组均明显增高（P〈0．01，n=5）。结论瘢痕成纤维细胞可作为Sp1转染的靶细胞，Sp1基因可能是导致瘢痕异常增生的重要基因。Objective To explore the feasibility of transfecting recombinant Spl into hypertrophic scar fibroblasts and investigate the proliferation and collagen Ⅰ , Ⅲ synthesis in the transfected cells. Methods Recombinant human Spl was transfected into hypertrophic scar fibroblasts with the karyocyte expressive vector. The expression of Spl, collagen Ⅰ , Ⅲ mRNA was tested by real time PCR. The change of cell proliferation was observed with CCK8 colorimeter. Results About 30% of transfected hypertrophic scar fibroblasts showed green fluorescence positive. The relative expression of Spl mRNA in transfected cells, empty-vector cell or untransfected cells group was 5.26 ± 0.76, 1.08 ± 0.18, 1.09 ± 0.15, respectively, showing a significant difference between thansfected and untransfected cells or between the transfected cells and empty-vector group （ P 〈 0.01, n = 5）, Expression of collagen Ⅰ , Ⅲ mRNA was 2.49 ± 0.40 and 1.88± 0.30 in transfected cells, 0.96±0.18 and 0.95 ± 0.18 in empty-vector cell,and 0.97 ± 0.15 and 0.93 ± 0.13 in untransfected cells, respectively, showing a significant difference between thansfected and untransfected cells or between the transfected cells and empty-vector group （ P 〈 0.01, n = 5 ）. Conclusions The hypertrophic scar fibroblasts could be as the target cells of Spl gene transfection. Spl gene may play an important role in abnormal collagen metabolism in hypertrophic scar.

关 键 词：转录因子 转染 瘢痕 成纤维细胞 

分 类 号：R622[医药卫生—整形外科]