Astragaloside IV attenuates glycated albumin-induced epithelial-to-mesenchymal transition by inhibiting oxidative stress in renal proximal tubular cells
Astragaloside IV attenuates glycated albumin-induced epithelial-to-mesenchymal transition by inhibiting oxidative stress in renal proximal tubular cells
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DOI：
10.1007/s12192-013-0438-7
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发表时间：
2014-01-01
影响因子：
3.8
通讯作者：
Gu, Yong
中科院分区：
文献类型：
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作者：
Qi, Weiwei;Niu, Jianying;Gu, Yong
在糖尿病肾病（DKD）中，上皮-间质转化（EMT）是肾小管损伤的典型病理过程。氧化应激被认为在DKD中起重要作用。黄芪甲苷（Astragaloside IV, a -IV）是黄芪的主要活性成分之一，具有广泛的生物活性。然而，A-IV对小管细胞EMT的调节作用尚不清楚。本研究旨在确定A-IV是否可以通过抑制氧化应激来减弱GA诱导的NRK-52E细胞系的EMT。CCK-8检测GA和a - iv诱导的细胞毒性。采用H2DCFDA检测细胞间活性氧（ROS）水平。通过添加外源NADPH氧化酶检测NADPH氧化酶活性，通过NBT检测超氧化物歧化酶（SOD）单位。我们用显微镜观察了NRK-52E细胞系的形态。我们进行了伤口愈合实验来测量细胞的流动性。为了检测α - sma和E-cadherin mRNA和蛋白的表达，我们采用实时聚合酶链反应（real-time PCR）、免疫荧光和western blot分析。A-IV显著减弱ga诱导的ROS扩增，降低升高的NADPH氧化酶活性水平，升高降低的SOD单位水平。ga诱导的NRK-52E细胞系在mRNA和蛋白水平上显示a-SMA表达增加，E-cadherin表达降低，而A-IV则减轻了a-SMA表达，增加了E-cadherin表达。我们的数据表明，GA可以通过氧化应激诱导NRK-52E细胞株EMT。这种作用可以通过A-IV对受损氧化还原平衡的调节而减弱。
In diabetic kidney disease (DKD), epithelial-to-mesenchymal transition (EMT) is a classic pathological process in tubular damage. Oxidative stress is considered to play an important role in DKD. Astragaloside IV (A-IV), one of the main active ingredients of Astragalus membranaceus, exhibits a wide range of biological activities. However, the effect of A-IV on regulating EMT in tubular cells is unclear. This study aims to determine whether A-IV could attenuate glycated albumin (GA)-induced EMT in the NRK-52E cell line by inhibiting oxidative stress. GA and A-IV-induced cytotoxicity were assayed by CCK-8. The intercellular reactive oxygen species (ROS) level was detected by H2DCFDA. The activity of NADPH oxidase was assayed by adding exogenous NADPH oxidase, and the superoxide dismutase (SOD) units were observed by NBT. We used a microscope to examine the morphology of the NRK-52E cell line. We conducted a wound healing assay to measure cell mobility. To determine mRNA and protein expressions of alpha-SMA and E-cadherin, we used real-time polymerase chain reaction (real-time PCR), immunofluorescence, and western blot analysis. A-IV significantly attenuated GA-induced amplification of ROS, lowered the increased level of NADPH oxidase activity, and elevated the decreased level of SOD units. The GA-induced NRK-52E cell line showed increased expression of a-SMA and decreased expression of E-cadherin in mRNA and protein levels, whereas A-IV alleviated the expression of a-SMA and increased the expression of E-cadherin. Our data demonstrate that GA could induce NRK-52E cell line EMT through oxidative stress. This effect could be attenuated by A-IV via regulation of the impaired redox balance.