GSTA3 Attenuates Renal Interstitial Fibrosis by Inhibiting TGF-Beta-Induced Tubular Epithelial-Mesenchymal Transition and Fibronectin Expression.
GSTA3 Attenuates Renal Interstitial Fibrosis by Inhibiting TGF-Beta-Induced Tubular Epithelial-Mesenchymal Transition and Fibronectin Expression.
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GSTA3 通过抑制 TGF-β 诱导的管状上皮-间质转化和纤连蛋白表达来减轻肾间质纤维化
DOI：
10.1371/journal.pone.0160855
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Tao L
中科院分区：
文献类型：
--
作者：
Xiao Y;Liu J;Peng Y;Xiong X;Huang L;Yang H;Zhang J;Tao L
小管上皮-间质转化（EMT）已被广泛认为是肾间质纤维化（RIF）的潜在机制。活性氧（ROS）的产生在管状EMT过程中起着至关重要的作用。本研究的目的是探讨tgf - β诱导EMT的相关分子机制，并确定谷胱甘肽s -转移酶- 3 （GSTA3）在这一过程中的潜在作用。通过iTRAQ筛选，确定单侧输尿管梗阻（UUO）大鼠的蛋白变化。采用免疫组化方法检测梗阻性肾病患者和UUO大鼠GSTA3蛋白的表达。采用Western blot和RT-PCR检测UUO大鼠和NRK-52E细胞中GSTA3蛋白和mRNA的表达。特异性转染siRNA和过表达质粒，评估GSTA3在RIF中的作用。用二氯荧光素荧光法测定活性氧的生成。UUO大鼠GSTA3蛋白和mRNA表达明显降低。免疫组化分析显示，UUO大鼠和梗阻性肾病患者肾皮质GSTA3表达降低。TGF-β1处理可下调NRK-52E细胞中GSTA3的表达，并与E-cadherin、megalin表达降低，α-平滑肌肌动蛋白表达升高相关。此外，敲低NRK-52细胞中的GSTA3可导致ROS和管状EMT的产生增加，而过表达GSTA3可改善ROS的产生并阻止管状EMT的发生。GSTA3在肾纤维化中对小管EMT起保护作用，提示GSTA3是RIF的潜在治疗靶点。
Tubular epithelial-mesenchymal transition (EMT) has been widely accepted as the underlying mechanisms of renal interstitial fibrosis (RIF). The production of reactive oxygen species (ROS) plays a vital role in tubular EMT process. The purpose of this study was to investigate the involved molecular mechanisms in TGF-beta-induced EMT and identify the potential role of glutathione S-transferase alpha 3 (GSTA3) in this process. The iTRAQ screening was performed to identify protein alterations of the rats underwent unilateral-ureteral obstruction (UUO). Protein expression of GSTA3 in patients with obstructive nephropathy and UUO rats was detected by immunohistochemistry. Protein and mRNA expression of GSTA3 in UUO rats and NRK-52E cells were determined by Western blot and RT-PCR. siRNA and overexpression plasmid were transfected specifically to assess the role of GSTA3 in RIF. The generation of ROS was measured by dichlorofluorescein fluorescence analysis. GSTA3 protein and mRNA expression was significantly reduced in UUO rats. Immunohistochemical analysis revealed that GSTA3 expression was reduced in renal cortex in UUO rats and patients with obstructive nephropathy. Treating with TGF-β1 down-regulated GSTA3 expression in NRK-52E cells, which have been found to be correlated with the decreased expression in E-cadherin and megalin and increased expression in α-smooth muscle actin. Furthermore, knocking down GSTA3 in NRK-52 cells led to increased production of ROS and tubular EMT, whereas overexpressing GSTA3 ameliorated ROS production and prevented the occurrence of tubular EMT. GSTA3 plays a protective role against tubular EMT in renal fibrosis, suggesting GSTA3 is a potential therapeutic target for RIF.