姜黄素对单侧输尿管梗阻大鼠肾小管上皮细胞转分化及TGF-β/Smads信号转导途径的影响

作者	单位
牛建昭	北京中医药大学发展规划处
李彧	北京中医药大学发展规划处
李亚东	北京中医药大学发展规划处
李健	北京中医药大学发展规划处
陈朝青	北京中医药大学发展规划处

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中文摘要:

目的观察姜黄素对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠肾小管上皮细胞转分化(epithelial-mesenchymal transition,EMT)及TGF-β/Smads信号转导途径的影响。方法建立大鼠UUO模型,分为假手术组、模型组及姜黄素1、2组。姜黄素1组术后第2天开始给予姜黄素100mg/(kg·d)灌胃,姜黄素2组术后第10天开始给予姜黄素100mg/(kg·d)灌胃。假手术组和模型组给予同等剂量的生理盐水灌胃。术后28天处死大鼠,取梗阻侧肾组织,免疫组化染色观察α-平滑肌肌动蛋白(α-smoothmuscle actin,α-SMA),E-钙黏蛋白(E-cadherin)的表达分布;Western blot检测转化生长因子-β1(transforminggrowth factor-β1,TGF-β1)、P-Smad2、P-Smad3及Smad7的蛋白表达,实时荧光定量PCR方法检测TGF-β1、转化生长因子-β受体Ⅰ型(tansforming growth factor-βreceptorⅠ,TβRⅠ)、胶原蛋白-Ⅰ(collagen-Ⅰ,Col-Ⅰ)和胶原蛋白-Ⅲ(collagen-Ⅲ,Col-Ⅲ)mRNA水平表达。结果 UUO组大鼠肾组织中α-SMA表达明显增强(P<0·01),E-cadherin表达明显减弱(P<0·01),P-Smad2和P-Smad3蛋白,TGF-β1蛋白及mRNA,TβRⅠmRNA及Col-Ⅰ、Col-ⅢmRNA的表达水平均明显上调(P<0·01),Smad7蛋白表达明显减少(P<0·01)。姜黄素1、2组α-SMA蛋白,P-Smad2及P-Smad3蛋白,TGF-β1蛋白及mRNA,TβRⅠmRNA,Col-Ⅰ、Col-ⅢmRNA的表达水平与UUO组比较显著下降(P<0·05,P<0·01),E-cadherin和Smad7蛋白表达显著升高(P<0·05,P<0·01)。结论姜黄素可干预UUO大鼠TGF-β/Smads信号通路中多个位点,抑制肾小管上皮细胞EMT的发生,从而减轻肾小管间质纤维化。

英文摘要:

Objective To observe the effects of curcumin on the epithelial mesenchymal transition(EMT) and TGF-β/Smads signal pathway in unilateral ureteral obstruction(UUO) rats.Methods UUO rat model was established.Rats were divided into the sham-operation group,the model group,the 1st curcumin group,and the 2nd curcumin group,respectively.From the 2nd day after surgery,rats of the 1st curcumin group were administrated with curcumin by gastrogavage at the daily dose of 100 mg/kg.From the 10th day after surgery,rats of the 2nd curcumin group were administrated with curcumin by gastrogavage at the daily dose of 100 mg/kg.Equal volume of normal saline was given to rats by gastrogavage in the sham-operation group and the model group.On the 28th day after surgery rats were killed.The renal tissues of the obstructed side were taken out.The distribution of mice α-smooth muscle actin(α-SMA) and E-cadherin expressions were observed using immunohistochemical method.Transforming growth factor-β1(TGF-β1),P-Smad2,P-Smad3,and Smad7 expressions were detected by Western blot.TGF-β1,transforming growth factor-β receptor Ⅰ(TβRⅠ),collagen-Ⅰ(Col-Ⅰ),and collagen-Ⅲ(Col-Ⅲ) mRNA expressions were detected by Real-time fluorescent quantitative PCR.Results α-SMA expressions were obviously enhanced,E-cadherin expressions obviously weakened,the protein expression levels of P-Smad2 and P-Smad3,protein and mRNA expression levels of TGF-β1,TβRⅠ mRNA,Col-Ⅰ mRNA,and Col-Ⅲ mRNA obviously up-regulated,Smad7 protein expressions obviously decreased(all P<0.01) in the renal tissues of rats in the UUO group.In the 1st curcumin group and the 2nd curcumin group,the protein expressions of α-SMA,P-Smad2,P-Smad3,and TGF-β1;mRNA expression levels of TGF-β1,TβRⅠ mRNA,Col-Ⅰ,and Col-Ⅲ obviously decreased(P<0.05,P<0.01),while the protein expressions of E-cadherin and Smad7 obviously increased(P<0.05,P<0.01).Conclusions Curcumin could intervene several sites of the TGF-β/Smads signal transduction pathway of UUO rats,inhibit the occurrence of EMT of renal tubular epithelial cells,thus attenuating the tubulo-interstitial fibrosis.

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