Membrane rafts-redox signalling pathway contributes to renal fibrosis via modulation of the renal tubular epithelial-mesenchymal transition
Membrane rafts-redox signalling pathway contributes to renal fibrosis via modulation of the renal tubular epithelial-mesenchymal transition
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DOI：
10.1113/jp275952
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发表时间：
2018-08-15
影响因子：
5.5
通讯作者：
Gao, Ping-Jin
中科院分区：
文献类型：
--
作者：
Han, Wei-Qing;Xu, Lian;Gao, Ping-Jin
膜筏（membrane rafts，MR）-氧化还原途径的特征是NADPH氧化酶亚基聚集并通过溶酶体融合激活、V型质子ATP酶亚基E2（Atp 6v 1 e2基因编码）转位和鞘磷脂磷酸二酯酶1（SMPD 1，SMPD 1基因编码）激活。在本研究中，我们假设MRs-氧化还原-衍生的活性氧（ROS）通过促进肾小管上皮-间质转化（EMT）参与肾脏炎症和纤维化。结果表明，转化生长因子-1（TGF-1）急性诱导大鼠肾小管细胞系NRK-52 E细胞中MR形成和ROS产生。此外，转染Atp 6v 1 e2小发夹RNA（shRNA）和SMPD 1 shRNA减弱TGF-1诱导的NRK-52 E细胞EMT标志物，包括E-钙粘蛋白，平滑肌肌动蛋白（SMA）和成纤维细胞特异性蛋白-1（FSP-1）的变化。此外，Erk 1/2活化可能是MRS氧化还原衍生的ROS的下游调节剂，因为两种shRNA均显著抑制TGF-1诱导的Erk 1/2磷酸化。进一步的体内研究表明，在血管紧张素II（AngII）诱导的高血压中，肾小管MR-氧化还原信号通路被激活，如在分离的肾小管细胞中MR结构域中NADPH氧化酶亚基Nox 4分数增加、SMPD 1激活和ROS含量增加所示。最后，肾脏转染Atp 6v 1 e2 shRNA和SMPD 1 shRNA显着防止肾纤维化和炎症，如所示的减少-SMA，纤连蛋白，胶原蛋白I，单核细胞趋化蛋白-1（MCP-1），细胞间粘附分子-1（ICAM-1）和肿瘤坏死因子-（TNF-α）在肾血管紧张素II输注大鼠。结论：MRS-氧化还原信号通路参与TGF-1诱导的肾小管EMT和AngII诱导的高血压中的肾脏炎症/纤维化。
The membrane rafts (MRs)-redox pathway is characterized by NADPH oxidase subunit clustering and activation through lysosome fusion, V-type proton ATPase subunit E2 (encoded by the Atp6v1e2 gene) translocation and sphingomyelin phosphodiesterase 1 (SMPD1, encoded by the SMPD1 gene) activation. In the present study, we hypothesized that the MRs-redox-derived reactive oxygen species (ROS) are involved in renal inflammation and fibrosis by promoting renal tubular epithelial-mesenchymal transition (EMT). Results show that transforming growth factor-1 (TGF-1) acutely induced MR formation and ROS production in NRK-52E cells, a rat renal tubular cell line. In addition, transfection of Atp6v1e2 small hairpin RNAs (shRNA) and SMPD1 shRNA attenuated TGF-1-induced changes in EMT markers, including E-cadherin, -smooth muscle actin (-SMA) and fibroblast-specific protein-1 (FSP-1) in NRK-52E cells. Moreover, Erk1/2 activation may be a downstream regulator of the MRs-redox-derived ROS, because both shRNAs significantly inhibited TGF-1-induced Erk1/2 phosphorylation. Further in vivo study shows that the renal tubular the MRs-redox signalling pathway was activated in angiotensin II (AngII)-induced hypertension, as indicated by the increased NADPH oxidase subunit Nox4 fraction in the MR domain, SMPD1 activation and increased ROS content in isolated renal tubular cells. Finally, renal transfection of Atp6v1e2 shRNA and SMPD1 shRNA significantly prevented renal fibrosis and inflammation, as indicated by the decrease of -SMA, fibronectin, collagen I, monocyte chemoattractant protein-1 (MCP-1), intercellular cell adhesion molecule-1 (ICAM-1) and tumour necrosis factor- (TNF-) in kidneys from AngII-infused rats. It was concluded that the the MRs-redox signalling pathway is involved in TGF-1-induced renal tubular EMT and renal inflammation/fibrosis in AngII-induced hypertension.