Respiratory Syncytial Virus Induces Oxidative Stress by Modulating Antioxidant Enzymes
Respiratory Syncytial Virus Induces Oxidative Stress by Modulating Antioxidant Enzymes
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DOI：
10.1165/rcmb.2008-0330oc
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发表时间：
2009-09-01
影响因子：
6.4
通讯作者：
Casola, Antonella
中科院分区：
文献类型：
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作者：
Hosakote, Yashoda M.;Liu, Tianshuang;Casola, Antonella
氧化应激在肺部炎症的发病机制中起重要作用。呼吸道合胞病毒（Respiratory syncytial virus，RSV）感染在体外可诱导活性氧（reactive oxygen species，ROS）的产生，在体内可诱导肺组织氧化损伤，但RSV诱导细胞氧化应激的机制尚不清楚。因此，我们确定呼吸道合胞病毒感染的气道上皮细胞是否改变了抗氧化酶（AOE）的表达和/或活性。用RSV感染A549细胞（人肺泡II型样上皮细胞系）和小气道上皮（SAE）细胞（来自终末细支气管的正常人细胞），并在不同时间点收获，以通过酶联免疫吸附测定法测量F-2-8异前列腺素，并通过比色测定法测量总谷胱甘肽和还原型谷胱甘肽（GSH和GSSG）。超氧化物歧化酶（SOD）1，2，和3，过氧化氢酶，谷胱甘肽过氧化物酶（GPx），谷胱甘肽S-转移酶（GST）的表达通过定量实时PCR和Western印迹法测定，并通过比色测定其活性。RSV感染诱导脂质过氧化产物的显着增加，以及GSH/GSSG比值显着下降。与未感染细胞相比，RSV感染细胞中SOD 1、SOD 3、过氧化氢酶和GST表达显著降低，同时SOD 2表达增加。RSV感染后，SOD总活性升高，而过氧化氢酶、GPx和GST活性降低。我们的研究结果表明，RSV诱导的细胞氧化损伤是ROS产生和抗氧化细胞防御之间不平衡的结果。氧化应激的调节是一种潜在的新的药理学方法，以改善RSV诱导的急性肺部炎症。
Oxidative stress plays an important role in the pathogenesis of lung inflammation. Respiratory syncytial virus (RSV) infection induces reactive oxygen species (ROS) production in vitro and oxidative injury in lungs in vivo, however, the mechanism of RSV-induced cellular oxidative stress has not been investigated. Therefore, we determined whether RSV infection of airway epithelial cells modified the expression and/or activities of antioxidant enzymes (AOE). A549 cells, a human alveolar type II-like epithelial cell line, and small airway epithelial (SAE) cells, normal human cells derived from terminal bronchioli, were infected with RSV and harvested at various time points to measure F-2-8 isoprostanes by enzyme-linked immunosorbent assay and total and reduced glutathione (GSH and GSSG) by colorimetric assay. Superoxide dismutase (SOD) 1, 2, and 3, catalase, glutathione peroxidase (GPx), and glutathione S-transferase (GST) expression was determined by quantitative real-time PCR and Western blot, and their activity was measured by colorimetric assays. RSV infection induced a significant increase of lipid peroxidation products as well as a significant decrease in the GSH/GSSG ratio. There was a significant decrease in SOD 1, SOD 3, catalase, and GST expression with a concomitant increase of SOD 2 in RSV-infected cells, compared with uninfected cells. Total SOD activity was increased, but catalase, GPx, and GST activities were decreased, after RSV infection. Our findings suggest that RSV-induced cellular oxidative damage is the result of an imbalance between ROS production and antioxidant cellular defenses. Modulation of oxidative stress represents a potential novel pharmacologic approach to ameliorate RSV-induced acute lung inflammation.