中华皮肤科杂志 ›› 2006, Vol. 39 ›› Issue (1): 19-21.

• 论著 • 上一篇    下一篇

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

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   

  1. 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
  • 收稿日期:2005-06-03 发布日期:2006-01-15
  • 基金资助:
    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)

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

XIAO Rong1, LUO Jing-ying1, LIU Fu-you2, ZHOU Ying1, LI Ya-ping1, ZHOU Yi-xin1, SU Yu-wen1, WEN Hai-quan1, YANG Xin-jie3   

  1. Department of Dermatology, the Second Xiangya Hospital of Central South University, Changsha 410011, China
  • Received:2005-06-03 Published:2006-01-15

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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CTGF mRNA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CTGF mRNA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.

关键词: RNA,小分子干扰, 硬皮病,系统性, 成纤维细胞, 结缔组织生长因子

Abstract: 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 semiquantitative 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.

Key words: RNA,small interfering, Scleroderma,systemic, Fibroblast, Connective tissue growth factor