RNA干扰对系统性硬化病皮肤成纤维细胞结缔组织生长因子表达的影响  

The effect of RNA interference on the expression of CTGF in skin fibroblasts of systemic sclerosis

作　　者：肖嵘[1] 罗婧莹[1] 刘伏友[2] 周英[1] 李亚萍[1] 周怡新[1] 苏玉文[1] 文海泉[1] 杨心洁[3] 

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

出　　处：《中华皮肤科杂志》2006年第1期19-21,共3页Chinese Journal of Dermatology

基　　金：湖南省卫生厅科研基金(B2003-051);湖南省发展计划委员会(湘计高技[2003]790-15)

摘　　要：目的研究RNA干扰对系统性硬化病(SS)皮肤成纤维细胞结缔组织生长因子(CTGF)mRNA和蛋白表达水平的影响。方法设计4个针对CTGF的特异性小干扰RNA(siRNA)及一个非特异性siRNA。体外转录获得siRNA,将siRNA用6-羧基荧光素(FAM)标记后瞬时转染至原代培养的SS患者皮肤成纤维细胞;转染48h后,以RT-PCR及蛋白印迹法分别检测CTGFmRNA及蛋白量的变化。结果转染了4个特异性siRNA的成纤维细胞中CTGFmRNA水平均有不同程度下调(P<0.001),抑制效果由强至弱为,siRNA742>siRNA828>siRNA578>siRNA948。蛋白印迹则显示有3个siRNA能使CTGF蛋白表达水平不同程度下调(P<0.001),其中siRNA742的抑制效应最强。结论RNA干扰能显著抑制SS成纤维细胞CTGF的表达。Objective To study the effect of RNA interference on the expression of CTGF in skin fibroblasts of systemic sclerosis （SS）. Methods Four CTGF specific siRNAs and a negative control siRNA were designed and then synthesized by in vitro transcription, siRNAs labeled with carboxyfluorescein-6-succimidyl ester （FAM） were transiently transfected into SS skin fibroblasts. Forty-eight hours after the fibroblasts were treated with siRNAs, the mRNA and protein expression of CTGF was detected by semiquantitatire RT-PCR and Western blot analysis, respectively. Results The mRNA and protein expression of CTGF in fibroblasts was significantly down-regulated by 4 and 3 CTGF specific siRNAs （both P 〈 0.001 ）, respectively, at different levels. Conclusion RNA interference could effectively suppress the expression of CTGF in SS skin fibroblasts.

关 键 词：RNA 小分子干扰 硬发病 系统性 成纤维细胞 结缔组织生长因子 

分 类 号：R593.2[医药卫生—内科学]