姜黄素对单侧输尿管梗阻大鼠上皮-间质转化及TGF-β/Smads信号通路的影响

[Effects of curcumin on the epithelial mesenchymal transition and TGF-beta/Smads signaling pathway in unilateral ureteral obstruction rats].

作者信息

Li Yu, Chen Zhao-qing, Li Ya-dong

机构信息

Department of Development Plan, Beijing University of Chinese Medicine, Beijing 100029.

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2011 Sep;31(9):1224-8.

PMID:22013801

Abstract

OBJECTIVE

To observe the effects of curcumin on the epithelial mesenchymal transition (EMT) and TGF-beta/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 alpha-smooth muscle actin (alpha-SMA) and E-cadherin expressions were observed using immunohistochemical method. Transforming growth factor-beta 1 (TGF-beta 1), P-Smad2, P-Smad3, and Smad7 expressions were detected by Western blot. TGF-beta 1, transforming growth factor-beta receptor I (TbetaRI), collagen-I (Col-I), and collagen-III (Col-III) mRNA expressions were detected by Real-time fluorescent quantitative PCR.

RESULTS

alpha-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-beta 1, TbetaRI 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 alpha-SMA, P-Smad2, P-Smad3, and TGF-beta 1; mRNA expression levels of TGF-beta 1, TbetaRI 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-beta/Smads signal transduction pathway of UUO rats, inhibit the occurrence of EMT of renal tubular epithelial cells, thus attenuating the tubulo-interstitial fibrosis.

摘要

目的

观察姜黄素对单侧输尿管梗阻（UUO）大鼠上皮-间质转化（EMT）及转化生长因子-β（TGF-β）/Smads信号通路的影响。

方法

建立UUO大鼠模型。将大鼠分为假手术组、模型组、姜黄素1组和姜黄素2组。术后第2天起，姜黄素1组大鼠每日经胃灌胃给予姜黄素，剂量为100 mg/kg。术后第10天起，姜黄素2组大鼠每日经胃灌胃给予姜黄素，剂量为100 mg/kg。假手术组和模型组大鼠经胃灌胃给予等体积生理盐水。术后第28天处死大鼠，取出梗阻侧肾组织。采用免疫组织化学方法观察小鼠α-平滑肌肌动蛋白（α-SMA）和E-钙黏蛋白表达分布。采用蛋白质印迹法检测转化生长因子-β1（TGF-β1）、磷酸化Smad2（P-Smad2）、磷酸化Smad3（P-Smad3）和Smad7表达。采用实时荧光定量PCR检测TGF-β1、转化生长因子-β受体I（TβRI）、I型胶原（Col-I）和III型胶原（Col-III）mRNA表达。

结果

UUO组大鼠肾组织中α-SMA表达明显增强，E-钙黏蛋白表达明显减弱，P-Smad2和P-Smad3蛋白表达水平、TGF-β1蛋白及mRNA表达水平、TβRI mRNA、Col-I mRNA和Col-III mRNA明显上调，Smad7蛋白表达明显降低（均P<0.01）。姜黄素1组和姜黄素2组中，α-SMA、P-Smad2、P-Smad3和TGF-β1蛋白表达；TGF-β1、TβRI mRNA、Col-I和Col-III mRNA表达水平明显降低（P<0.05，P<0.01），而E-钙黏蛋白和Smad7蛋白表达明显增加（P<0.05，P<0.01）。