Advanced oxidation protein products induce hepatocyte epithelial-mesenchymal transition via a ROS-dependent, TGF-/Smad signaling pathway
Advanced oxidation protein products induce hepatocyte epithelial-mesenchymal transition via a ROS-dependent, TGF-/Smad signaling pathway
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高级氧化蛋白产品通过 ROS 依赖性 TGF-/Smad 信号通路诱导肝细胞上皮间质转化
DOI：
10.1002/cbin.10792
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发表时间：
2017
影响因子：
3.9
通讯作者：
Zhou Jie
中科院分区：
文献类型：
--
作者：
Sun Shibo;Xie Fang;Zhang Qifan;Cui Zhonglin;Cheng Xinsheng;Zhong Feng;He Kun;Zhou Jie
上皮-间充质转化(EMT)发生在慢性肝损伤后肝纤维化的发展过程中。然而，肝细胞EMT调控的分子机制仍不清楚。本研究的目的是确定晚期氧化蛋白产物(AOPP)是否对肝细胞EMT有影响。在体外和体内实验中，用AOPP处理人L02肝细胞系和正常SD大鼠的肝细胞。用Western blotting和定量逆转录聚合酶链式反应(qRT-PCR)检测L02肝细胞EMT细胞和分子标志物的表达。用创面愈合试验分析肝细胞的迁移能力。用二氯荧光素(DCF)法检测细胞内活性氧(ROS)。肝组织切片用免疫组织化学方法检测EMT标志物的表达，用Masson‘s三色染色观察胶原蛋白的表达。结果表明，AOPP可导致肝细胞发生EMT，其机制可能与E-钙粘蛋白表达减少、波形蛋白表达增加、胶原蛋白沉积增加以及促进细胞在体内和体外迁移有关。AOPP还能促进L02细胞的迁移，促进ROS的产生，激活转化生长因子-β受体和Smad信号通路。抑制细胞内ROS的产生和阻断转化生长因子-β受体可逆转AOPP诱导的肝细胞内皮细胞转化。本研究发现了一种新的调节肝细胞EMT的机制，这一发现可能对控制肝纤维化有一定的意义。
Epithelial–mesenchymal transition (EMT) occurs during the progression of liver fibrosis in response to chronic liver injury. However, the molecular mechanism underlying the regulation of hepatocyte EMT remains unclear. The aim of this study was to determine whether advanced oxidation protein products (AOPP) had an effect on hepatocyte EMT. The human L02 hepatocyte cell line and hepatocytes from normal Sprague–Dawley rats were challenged with AOPP treatment in both in vitro and in vivo studies. The expression of cell and molecular markers of EMT in L02 hepatocytes were studied using Western blotting, and quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR) assays. Hepatocyte migratory potential was analyzed using a wound healing assay. Intracellular reactive oxygen species (ROS) were detected using the dichlorofluorescein (DCF) assay. In liver tissue sections, expression of EMT markers was evaluated using immunohistochemistry, and collagen was assessed using histochemical staining with Masson's trichrome. The findings were that AOPP treatment resulted in EMT in hepatocytes, which was associated with reduced expression of E‐cadherin, increased expression of vimentin, increased deposition of collagen protein, and enhanced cell migration in vivo and in vitro. AOPP was also found to promote migration in L02 cells, and to promote the production of ROS and the activation of TGF‐βR and Smad signaling. Inhibition of the generation of intracellular ROS and TGF‐β receptor blocking could reverse AOPP‐induced EMT in hepatocytes. This study has identified a novel mechanism in the regulation of hepatocyte EMT, and the findings may have implications for the control of liver fibrosis.