高效转染体外转录合成的CTGF-siRNA至硬皮病皮肤成纤维细胞  

High efficiency transfection of CTGF-siRNA transcripted in vitro into scleroderma skin fibroblast

作　　者：肖嵘[1] 罗婧莹[1] 刘伏友[2] 杨心洁[3] 邱湘宁[1] 赵会平[1] 苏玉文[1] 文海泉[1] 

机构地区：[1]中南大学湘雅二医院皮肤性病科,湖南长沙410008 [2]中南大学湘雅二医院肾内科,湖南长沙410011 [3]中南大学湘雅二医院医学实验中心,湖南长沙410011

出　　处：《中国现代医学杂志》2005年第21期3248-3250,3254,共4页China Journal of Modern Medicine

基　　金：湖南省发展改革委员会资助(湘计高技[2003]790-15);湖南省卫生厅科研基金资助(B2003-051)

摘　　要：目的研究CTGF-siRNA的体外转录合成及使用专门转染试剂盒将其转染至硬皮病皮肤成纤维细胞的转染效率,为将RNA干扰技术深入应用到硬皮病治疗做铺垫。方法体外培养硬皮病皮肤成纤维细胞;设计CTGF的siRNA;通过体外转录获得siRNA;将siRNA以SilencerTMsiRNALabelingKit标记;用siPORTTMAmine转染至成纤维细胞;荧光显微镜观察转染情况。结果转染率可高达85.32%。结论该方法操作简便,且可达到理想的转染率。[Objective] To study the transcription of CTGF-siRNA in vitro and CTGF-siRNA transfection efficiency in scleroderma skin fibroblast with the kit used specially for transfecting siRNA, in order to do spade work for applying technology of RNA interference deeply to the treatment of scleroderma. [Methods] Cultured scleroderma lesional dermal fibroblast in vitro. Designed CTGF-siRNA, obtained it through transcripting and labeled it with Silencer^TMsiRNA Labeling Kit. siPOR^TM Amine was used in the transfection. Results of the transfection were detected through fluorescent microscope. [Results] Transfection efficiency could reach as high as 85.32%. [Conclusion] Our experiment was convenient and high transfection efficiency could be reached.

关 键 词：SIRNA 硬皮病 成纤维细胞 转染 

分 类 号：R593.25[医药卫生—内科学]