针对CTGF的shRNA抑制TGFβ1诱导人肾小管上皮细胞合成胶原

Inhibitory effects of shRNA targeting CTGF in transforming growth factor β1-induced collagen synthesis in human renal tubular epithelial cells

目的研究针对结缔组织生长因子(connectivetissuegrowthfactor,CTGF)的短发夹RNA(shorthairpinRNA,shRNA)对转化生长因子β1(TGFβ1)诱导人肾小管上皮细胞株(humanrenaltubularepithelialcell,HKC)合成胶原的抑制作用。方法设计和构建带有U6启动子的能产生针对CTGF的shRNA的RNA干扰表达质粒,转染至受TGFβ1刺激的HKC,以RT-PCR和Westernblot检测HKC转染前后CTGF表达,以3H脯氨酸掺入法检测总胶原合成量。结果和仅受TGFβ1刺激组相比,转染了RNA干扰表达质粒的受TGFβ1刺激的HKCCTGFmRNA和蛋白表达明显减弱,总胶原合成量下降(P<0.01)。结论针对CTGF的短发夹RNA能明显抑制TGFβ1诱导HKC合成胶原。

[Objective] To investigate the effect of short hairpin RNA （shRNA） targeting CTGF on synthesizing collagen ability of human renal tubular epithelial eells （HKC） cultured in vitro with transforming growth factor β1 （TGFβ1）. [Methods] RNA interference plasmid including U6 promoter that can express shRNA of CTGF was designed, constructed, and transfected into HKC cultured with TGFβ1 in vitro. CTGF expression was examined by reverse transcription polymerase chain reaction （RT-PCR） and western blot. Total collagen synthesis was investigated by means of 3H proline incorporation. [Results] After plasmid transfected, CTGF expression and total collagen synthesis were decreased signitieandy compared with that stimulated by only TGFβ1 （P〈0.01）. [Conclusion] The shRNA targeting CTGF could inhibit efficiently collagen synthesis in HKC cuhured in vitro with TGFβ1.