Expression of NADPH oxidases and enhanced H2O2-generating activity in human coronary artery endothelial cells upon induction with tumor necrosis factor-α
Expression of NADPH oxidases and enhanced H2O2-generating activity in human coronary artery endothelial cells upon induction with tumor necrosis factor-α
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DOI：
10.1016/j.intimp.2008.05.004
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发表时间：
2008-10-01
影响因子：
5.6
通讯作者：
Tsunawaki, Shohko
中科院分区：
文献类型：
--
作者：
Yoshida, Lucia S.;Tsunawaki, Shohko
肿瘤坏死因子(TNF)- α可增强活性氧（ROS）的产生，在川崎病冠状动脉炎和动脉瘤的发展中起着至关重要的作用。我们假设血管NADPH氧化酶（Nox）酶通过提高ROS的生成参与了tnf - α触发的内皮损伤。因此，我们在此研究了人类冠状动脉内皮细胞（HCAEC）中Nox酶的表达以及tnf - α对Nox介导的ROS生成的影响。我们发现HCAEC在培养中自发产生基础水平（0.53 nmol/min/mg蛋白）的H2O2。在寻找产生H2O2的Nox成分时，发现在HCAEC中表达了两种不同的Nox4亚型：原型Nox4A和较短的Nox4B，分别在核后上清液和核分数中表达。其他表达的Nox家族成分有：作为mrna， Nox4C、Nox4D、Nox1、p51（Nox）和Racs；作为mrna和蛋白，Nox2、p22（phox）、p47（phox）和p67（phox）。在培养物中加入tnf - α后，生成h2o2的活性增加了三倍，同时增强了Nox4A、p22（phox）、p47（phox）和p67（phox）蛋白的表达。综上所述，这些结果表明tnf - α赋能的HCAEC诱导Nox2和Nox4A酶产生ros活性增强，这可能在氧化应激引起的初始内皮功能障碍中起重要作用。(c) 2008 Elsevier B.V.版权所有
Tumor necrosis factor (TNF)-alpha, which potentiates reactive oxygen species (ROS) generation, is crucial for the development of coronary arteritis and aneurysm in Kawasaki disease. We hypothesized that vascular NADPH oxidase (Nox) enzymes participate in the TNF-alpha-triggered endothelial damage through elevating ROS generation. Thus, we herein examine the expression of Nox enzymes in human coronary artery endothelial cells (HCAEC) and the effects of TNF-alpha on Nox-mediated ROS generation. We show that HCAEC in culture spontaneously generate H2O2 at basal level (0.53 nmol/min/mg protein). In searching for Nox components responsible for the H2O2 generation, two distinct isoforms of Nox4 are found expressed in HCAEC: the prototype Nox4A and the shorter Nox4B, respectively in the postnuctear supernatant and the nuclear fractions. Other expressed Nox family components are: as mRNAs, Nox4C, Nox4D, Nox1, p51(nox), and Racs; as mRNAs and proteins, Nox2, p22(phox), p47(phox), and p67(phox). The H2O2-generating activity increases up to three-fold upon inclusion of TNF-alpha in culture, concomitantly with augmented expressions of Nox4A, p22(phox), p47(phox) and p67(phox) proteins. Together, these results suggest that Nox2 and Nox4A enzymes are induced by TNF-alpha endowing HCAEC with enhanced ROS-generating activity, which may play a rote in the initial endothelial dysfunction through oxidative stress. (c) 2008 Elsevier B.V. All rights reserved.