首页> 美国卫生研究院文献>The Journal of Biological Chemistry >Thioredoxin-interacting Protein Mediates High Glucose-induced Reactive Oxygen Species Generation by Mitochondria and the NADPH Oxidase Nox4 in Mesangial Cells
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Thioredoxin-interacting Protein Mediates High Glucose-induced Reactive Oxygen Species Generation by Mitochondria and the NADPH Oxidase Nox4 in Mesangial Cells

机译:硫氧还蛋白相互作用蛋白介导肾小球系膜细胞中线粒体和NADPH氧化酶Nox4产生的高糖诱导的活性氧。

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摘要

Thioredoxin-interacting protein (TxNIP) is up-regulated by high glucose and is associated with oxidative stress. It has been implicated in hyperglycemia-induced β-cell dysfunction and apoptosis. As high glucose and oxidative stress mediate diabetic nephropathy (DN), the contribution of TxNIP was investigated in renal mesangial cell reactive oxygen species (ROS) generation and collagen synthesis. To determine the role of TxNIP, mouse mesangial cells (MC) cultured from wild-type C3H and TxNIP-deficient Hcb-19 mice were incubated in HG. Confocal microscopy was used to measure total and mitochondrial ROS production (DCF and MitoSOX) and collagen IV. Trx and NADPH oxidase activities were assayed and NADPH oxidase isoforms, Nox2 and Nox4, and antioxidant enzymes were determined by immunoblotting. C3H MC exposed to HG elicited a significant increase in cellular and mitochondrial ROS as well as Nox4 protein expression and NADPH oxidase activation, whereas Hcb-19 MC showed no response. Trx activity was attenuated by HG only in C3H MC. These defects in Hcb-19 MC were not due to increased antioxidant enzymes or scavenging of ROS, but associated with decreased ROS generation. Adenovirus-mediated overexpression of TxNIP in Hcb-19 MC and TxNIP knockdown with siRNA in C3H confirmed the specific role of TxNIP. Collagen IV accumulation in HG was markedly reduced in Hcb-19 cells. TxNIP is a critical component of the HG-ROS signaling pathway, required for the induction of mitochondrial and total cell ROS and the NADPH oxidase isoform, Nox4. TxNIP is a potential target to prevent DN.
机译:硫氧还蛋白相互作用蛋白(TxNIP)被高葡萄糖上调,并与氧化应激相关。它与高血糖诱导的β细胞功能障碍和细胞凋亡有关。由于高葡萄糖和氧化应激介导糖尿病性肾病(DN),因此研究了TxNIP在肾系膜细胞活性氧(ROS)生成和胶原合成中的作用。为了确定TxNIP的作用,将从野生型C3H和TxNIP缺陷型Hcb-19小鼠培养的小鼠系膜细胞(MC)在HG中孵育。共聚焦显微镜用于测量总和线粒体ROS的产生(DCF和MitoSOX)和IV型胶原。测定Trx和NADPH氧化酶活性,并通过免疫印迹测定NADPH氧化酶亚型Nox2和Nox4,以及抗氧化酶。暴露于HG的C3H MC引起细胞和线粒体ROS的显着增加,以及Nox4蛋白表达和NADPH氧化酶的活化,而Hcb-19 MC无反应。 Trx活性仅在C3H MC中被HG减弱。 Hcb-19 MC中的这些缺陷不是由于抗氧化酶增加或ROS清除所致,而是与ROS产生减少有关。腺病毒介导的Hxb-19 MC中TxNIP的过表达和C3H中siRNA的TxNIP敲低证实了TxNIP的特定作用。 Hcb-19细胞中HG中胶原IV的积累明显减少。 TxNIP是HG-ROS信号通路的关键组成部分,是诱导线粒体和总细胞ROS和NADPH氧化酶同工型Nox4所必需的。 TxNIP是防止DN的潜在目标。

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