RNA干扰对人瘢痕疙瘩成纤维细胞TGFβ1刺激后胶原合成的影响.doc

RNA干扰对人瘢痕疙瘩成纤维细胞TGFβ1刺激后胶原合成的影响
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【摘要】目的体外构建结缔组织生长因子(CTGF)序列特异性小干扰RNA表达载体,转染导入后研究其对人瘢痕疙瘩成纤维细胞(HKFs)胶原蛋白分泌的影响。方法体外构建CTGF序列特异性小干扰RNA质粒表达载体,Dosper脂质体转染人瘢痕疙瘩成纤维细胞后,观察加入10 ng/mL剂量的转化生长因子(TGFβ1)刺激前后胶原蛋白水平的变化。结果转染小干扰RNA质粒后,%;经TGFβ1刺激后,蛋白表达仍无明显变化。结论小干扰RNA质粒表达载体可抑制瘢痕疙瘩成纤维细胞胶原蛋白分泌,为临床上瘢痕疙瘩的基因治疗提供理论依据。
【关键词】 RNA干扰;人瘢痕成纤维细胞;胶原蛋白;转化生长因子β1
Abstract: Objective To investigate the effect of small interfering RNA(siRNA) expressing plasmids targeting connective tissue growth factor(CTGF) on collagen secretion in human keloid fibroblasts(HKFs). Methods Constructed siRNA
expressing plasmids targeting CTGF were transfected into HKFs by Dosper package. The collagen level was measured by 3Hproline incorporation method before and after HKFs were stimulated by transforming growth factor(TGFβ1) of the dosage of 10 ng/mL. Results The collagen level was only % of the normal level and had no obvious change even after stimulated by TGFβ1. Conclusion The downregulation of collagen by constructed siRNAexpressing plasmids may be a new way to treat keloid.
Keywords: RNA interference; human keloid fibroblast; collagen; TGFβ1
瘢痕疙瘩(Keloid,KD)是皮肤损伤如创伤、烧伤或手术后引发的以胶原过度沉积于真皮和皮下组织为特征的过度瘢痕化,其发病机制是异常表达的转化生长因子(TGFβ1)通过结缔组织生长因子(CTGF)促进病理性瘢痕的形成[1]。已有研究表明,TGFβ是一类具有多种生物学功能的蛋白多肽类物质,在创伤愈合的所有阶段均发挥了重要作用[2]。RNA干扰(RNA interference,RNAi)是由双链RN