中文摘要：

目的：探讨肾气丸含药血清对转化生长因子β1（TGF-β1）诱导人肾小管上皮细胞株HK-2转分化的作用及其分子机制。方法：将细胞分为：无血清对照组,TGF-β1组（10ng/mL）,空白血清组,肾气丸低、中、高剂量组。观察各组细胞形态学变化,ELISA法检测细胞上清液中Ⅰ型胶原（Col-Ⅰ）和纤维连接蛋白（FN）含量变化;荧光定量PCR法检测E-cadherin和Vimentin mRNA水平,蛋白免疫印迹法检测Smad2/3、ILK、GSK-3β等蛋白及其磷酸化水平。结果：TGF-β1刺激后,HK-2细胞由铺路石变成长梭形状,E-cadherin基因及蛋白水平下调（P〈0.01）,Vimentin基因及蛋白水平显著上升（P〈0.01）;给予肾气丸含药血清干预后,可抑制上述改变（P〈0.01,P〈0.05）。进一步研究发现,肾气丸能显著抑制p-Smad2/3、ILK、p-GSK-3β和β-catenin蛋白表达（P〈0.05,P〈0.01）。结论：肾气丸能显著抑制TGF-β1诱导HK-2细胞转分化过程,作用机制可能与其抑制TGF-β/Smads/ILK信号通路的激活有关。

英文摘要：

Objective： To investigate the effects and molecular mechanism of Shenqi Pill containing serum on TGF-β1-induced renal tubular epithelial-mesenchymal transition（EMT） HK-2 cells. Methods： HK-2 cells were divided into the control group（non-serum）, TGF-β1 group（10ng/mL）, blank serum group and Shenqi Pill low, middle and high dose group. The morphological changes of cells were observed, and the levels of Col-I and FN were detected by ELISA assay. The mRNAexpression of E-cadherin and Vimentin mRNA were detected by Real-time PCR, and the protein expression and phosphorylation levels of Smad2/3, ILK, GSK-3β were detected by Western Blot. Results： After the stimulation of TGF-β1, the morphology of HK-2 cells transformed from paving stone into long spindle shapes. The gene and protein expression of E-cadherin were decreased（P〈0.01）, while vimentin significantly increased（P〈0.01）. But, Shenqi Pill containing serum inhibited the above changes（P〈0.01, P〈0.05）. Further research showed that Shenqi Pill significantly inhibited protein expression of p-Smad2/3, ILK, p-GSK-3β and β-catenin（P〈0.05, P〈0.01）. Conclusion： Shenqi Pill could significantly inhibit TGF-β1-induced renal tubular EMT HK-2 cells, whose mechanism maybe related to inhibit the activation of TGF-β/Smads/ILK signal pathway.