白藜芦醇对TGF-β1介导的肾小管上皮细胞表型转化和Hedgehog信号的影响  

Effect of resveratrol on TGF-β1-mediated epithelial-mesenchymal transition and hedgehog signaling in renal tubular epithelial cells

作　　者：王斯璐[1] 吴存造[2] 潘晓东[2] 洪炜龙[1] 林成成[1] 陈必成[1] 白永恒[1] 

机构地区：[1]温州医科大学附属第一医院外科实验室,325000 [2]温州医科大学附属第一医院移植科,325000

出　　处：《浙江医学》2015年第7期536-538,548,I0001,I0002,共6页Zhejiang Medical Journal

基　　金：浙江省自然科学基金(LQ12H05001;LY12H05004);温州市科技计划项目(Y20110028)

摘　　要：目的探讨白藜芦醇(Res)对转化生长因子-β1(TGF-β1)诱导的肾小管上皮细胞(NRK-52E)表型转化(EMT)的影响及其分子机制。方法体外培养肾小管上皮细胞,分为对照组、诱导组(TGF-β1:5μg/L)、干预1组(TGF-β1+Res10μmol/ml)和干预2组(TGF-β1+Res100μmol/ml)。采用细胞免疫荧光和qRT-PCR技术检测EMT、细胞外基质和Hedgehog信号相关分子表达水平。结果 TGF-β1作用NRK-52E细胞后,不仅促进其受体TGF-β1R表达(P<0.05),且肌成纤维细胞标志物(α-SMA)和基质成分Ⅲ型胶原的表达增加,上皮细胞标志物(E-cadherin)表达下降。应用Res干预后,不仅抑制甚至逆转了上述结果,且抑制了TGF-β1介导的MMP-2/TIMP-2比值下降(P<0.05),促进基质成分降解。此外,Res也抑制TGF-β1诱导的PCNA表达上调(P<0 05),以及增殖相关的Hedgehog信号(Shh、Ptch1和Gli 1)活化(P<0.05)。结论 Res可能通过拮抗Hedgehog信号来抑制TGF-β1所诱导的NRK-52E细胞增殖、EMT和细胞外基质累积。Objective To investigate the effect of resveratrol（RES） on TGF- β 1-mediated epithelial-mesenchymal transi- tion （EMT） in renal tubular epithelial cells and its molecular mechanism. Methods Renal tubular epithelial NRK-52E cells were divided into control group, TGF- β1 group （EMT was induced by 5μg/L TGF-β 1）, treatment group 1 and 2 （EMT-induced ceils were treated with res 10 or 100μmol/ml）. qRT-PCR and immunofluorescence staining were performed to evaluate the mRNA and protein expression of EMT, extracellular matrix, and Hedgehog signaling molecules. Results After treated with TGF-β1, NRK-52E cells were transited from epithelial phenotype to mesenchymal phenotype, accompanied with down-regulated expres- sion of epithelial marker （E-cadherin） and up-regulated expression of TGF- β1 R（P〈0.05）, mesenchymal markers （ α -SMA） and matrix components （Type III collagen）, which were inhibited or reversed by resveratrol treatment. In addition, resveratrol inhibited TGF-β 1-mediated down-regulation of MMP-2/TIMP-2, resulting in the degradation of matrix components. Resveratrol also abolished up-regulated expression of PCNA （P〈0.05）, and activation of hedgehog signaling （Shh, Ptchl and Gill） induced by TGF- β1 in NRK-52E cells（P〈0.05）. Conclusion Resveratrol can inhibit TGF-β 1-induced proliferation, EMT and matrix depo- sition in NRK-52E cells via the hedgehog pathway.

关 键 词：白藜芦醇 转化生长因子-Β1 表型转化 肾小管上皮细胞 Hedgehog信号 

分 类 号：R285[医药卫生—中药学]