摘要
目的探讨重组ANGPTL-1基因转染人皮肤成纤维细胞的可行性,以及转染转染后对细胞增殖速率和Ⅰ、Ⅲ型胶原合成的影响。方法体外重组ANGPTL-1基因,借助真核表达载体系统,转入体外培养的人皮肤成纤维细胞,流式细胞仪检测基因表达及细胞转染效率;RT-PCR比较转染前后ANGPTL-1 mRNA及Ⅰ、Ⅲ型胶原mRNA的表达。结果基因转染后,(63±7.1)%的被转染细胞表达目的基因;转染组ANGPTL-1 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);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)。结论人皮肤成纤维细胞可作为ANGPTL-1转染的靶细胞,ANGPTL-1基因可能是促进Ⅰ、Ⅲ型胶原合成的重要基因之一。
Objective To explore the feasibility of transfecting ANGPTL-1 genes into human skin fibroblasts and investigate the effects of transfecting ANGPTL-1 genes on proliferation and collegen I , Ill synthesis. Methods Recombinant human ANGPTL-1 was transfected into skin fibroblasts with the karyocyte expressive vector.The expression of ANGPTL-1 and collagen I, Ⅲ mRNA were quantified by RT- PCR. Results Transfected human skin fibroblasts showed green fluorescence positive nearly in 63±7.1%, while the untransfected and empty-vector group could not. Expression of ANGPTL-1 mRNA increased statistically significant in transfected cells compared with either untransfected or empty-vector group （P〈0.01,n=5）. Expression of collagen I , Ⅲ mRNA both increased statistically significant in transfected cells compared with either untransfected or empty-vector group （P〈0.01 ,n=5）. Conclusion The human skin fibroblasts seems to be the target cells of ANGPTL-1 gene transfer.ANGPTL-1 may be a critical factor for anti-aging.
出处
《组织工程与重建外科杂志》
2012年第6期323-325,共3页
Journal of Tissue Engineering and Reconstructive Surgery
