8-Nitroguanosine formation in viral pneumonia and its implication for pathogenesis
8-Nitroguanosine formation in viral pneumonia and its implication for pathogenesis
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DOI：
10.1073/pnas.0235623100
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发表时间：
2003-01-21
影响因子：
11.1
通讯作者：
Maeda, H
中科院分区：
文献类型：
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作者：
Akaike, T;Okamoto, S;Maeda, H
对于许多疾病，有人提出一氧化氮 (NO) 介导发病机制。在这项研究中，我们探索了 NO 诱导的病毒发病机制，重点关注体内 8-硝基鸟苷形成所证明的核酸损伤。诱导型一氧化氮合酶（iNOS）缺陷的野生型小鼠和同窝小鼠感染了流感或仙台病毒。使用 8-硝基鸟苷特异性抗体通过免疫组织化学方法评估病毒感染的肺中 8-硝基鸟苷的形成。该抗体可以轻松检测到用过氧亚硝酸盐处理的 RNA 或从培养的表达 iNOS 的 RAW 264 细胞中获得的 RNA 的广泛硝化。强的 8-硝基鸟苷免疫染色主要在病毒感染的野生型小鼠的支气管和细支气管上皮细胞的胞质溶胶中明显，但在 iNOS 缺陷小鼠中则不然。该染色与肺中的 NOS 免疫染色共定位。在抗体反应过程中添加外源真实的 8-硝基鸟苷后，以及用连二亚硫酸钠预处理组织后，8-硝基鸟苷染色消失，将 8-硝基鸟苷还原为 8-氨基鸟苷。野生型小鼠肺部产生过量的 NO，但 iNOS 缺陷小鼠在病毒感染后被消除；该结果也与 8-硝基鸟苷和 3-硝基酪氨酸的形成密切相关。 NOS表达缺乏的后果之一是肺部组织病理学变化和感染致死率显着改善，但不影响细胞因子反应和病毒清除。有趣的是，8-硝基鸟苷在体外显着刺激细胞色素 P450 还原酶和 MOS 产生超氧化物。目前的数据证明了体内 8-硝基鸟苷的形成，并表明 NO 诱导的硝化应激在病毒发病机制中的潜在作用。
For many diseases, mediation of pathogenesis by nitric oxide (NO) has been suggested. in this study, we explored NO-induced viral pathogenesis with a focus on nucleic acid damage as evidenced by 8-nitroguanosine formation in vivo. Wild-type mice and littermate mice deficient in inducible NO synthase (iNOS) were infected with influenza or Sendai virus. Formation of 8-nitroguanosine in virus-infected lungs was assessed immunohistochemically with an antibody specific for 8-nitroguanosine. Extensive nitration of RNA either treated with peroxynitrite or obtained from cultured RAW 264 cells expressing iNOS was readily detected by this antibody. Strong 8-nitroguanosine immunostaining was evident primarily in the cytosol of bronchial and bronchiolar epithelial cells of virus-infected wild-type mice but not iNOS-deficient mice. This staining colocalized with NOS immunostaining in the lung. 8-Nitroguanosine staining disappeared after addition of exogenous authentic 8-nitroguanosine during the antibody reaction and after pretreatment of tissues with sodium hydrosulfite, which reduces 8-nitroguanosine to 8-aminoguanosine. NO was generated in excess in lungs of wild-type mice but was eliminated in iNOS-deficient mice after virus infection; this result also correlated well with formation of 8-nitroguanosine and 3-nitrotyrosine. One consequence of the lack of NOS expression was marked improvement in histopathological changes in the lung and the lethality of the infection without effects on cytokine responses and viral clearance. it is intriguing that 8-nitroguanosine markedly stimulated superoxide generation from cytochrome P450 reductase and MOS in vitro. The present data constitute a demonstration of 8-nitroguanosine formation in vivo and suggest a potential role for NO-induced nitrative stress in viral pathogenesis.