Molecular Mechanisms of Redox-Regulation in Viral Pathogenesis

病毒发病机制中氧化还原调节的分子机制

• 批准号: 09470046
• 负责人: AKAIKE Takaaki
• 金额: $ 3.71万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470046/
• 关键词: viral pathogenesis; NO; oxidative stress; peroxynitrite; free radicals; viral mutation; viral evolution

The importance of free radical molecular species in the pathogenesis of various viral diseases has been increasingly recognized in recent years. Oxygen radicals such as superoxide (OィイD3-(/)2ィエD3) and hydroxyl radical (・OH) have been implicated as possible pathogenic molecules in viral disease pathogenesis. Much attention has been given to another simple inorganic radical, i.e., NO in the host's defense mechanism and pathogenesis of virus infection. The NO synthesis pathway, in particular the inducible isoform of NO synthase (iNOS), is expressed in different viral diseases via induction of proinflammatory cytokines such as interferon-γ and interleukin-1β. iNOS produces an excessive amount of NO for a long time compared with other constitutive isoforms of NOS, i.e., neuronal NOS and endothelial NOS. Our present results indicate that overproduction of NO and oxygen radicals should be a common phenomenon in various infections. Reactive nitrogen oxide species such as peroxynitrite are prod … More uced in biological systems through the reaction of NO with OィイD3-(/)2ィエD3. Peroxynitrite and its biological actions are of considerable interest in that peroxynitrite causes oxidation and nitration of amino acid residues of proteins and guanine of DNA, lipid peroxidation, and DNA cleavage. In fact, it is revealed in our current study that peroxynitrite is critically involved in the viral pathogenesis through its nonselective injury to the host'cells and tissues. Thus, understanding of the role of NO and oxygen radical generation in virus infections will provide insight into not only viral pathogenesis but also the host-pathogen interaction in microbial infections at a molecular level. Moreover, among various NO-induced pathological effects, the mutagenic potential of NO in microbial pathogens is also intriguing. For viral mutation, it is of potential interest to investigate a possible association of oxidative stress and virulence of viruses. In this context, our recent study verifies for the first time that oxidative stress induced by high-output NO accelerates RNA virus mutations. Briefly, by using a recombinant RNA virus, Sendai virus (a negative-sense and single-strand RNA virus), containing a marker gene for genetic mutation and iNOS knockout mice, we obtained solid and direct evidence showing that overproduction of NO in the hosts (wild-type mice) in vivo apparently increases and accelerates viral mutation rates compared with the situation in iNOS-deficient mice. This process of accelerated mutation expands the heterogeneity of variants of the pathogen, leading to rapid evolution under selective pressure. NO and OィイD3-(/)2ィエD3- and hence peroxynitrite generation occurs universally in infected hosts. This finding therefore has great implications for the RNA virus evolution in general, including the rapid generation of drug-resistant and immunologically tolerant and cell tropism-altered mutants of HIV in vivo. Less

近年来，自由基分子在各种病毒性疾病发病机制中的重要性日益被人们所认识。超氧阴离子自由基(O-ィイD3-(/)2-ィエD3)和羟基自由基(·OH)等可能是病毒致病的致病分子。另一种简单的无机自由基，即NO在宿主的防御机制和病毒感染的发病机制中受到了极大的关注。NO合成途径，尤其是诱导型一氧化氮合酶(INOS)，在不同的病毒疾病中通过诱导炎性细胞因子如干扰素-γ和白介素1-β表达。与一氧化氮合酶的其他组成亚型，即神经型一氧化氮合酶和内皮型一氧化氮合酶相比，一氧化氮合酶长期产生过量的一氧化氮。我们目前的结果表明，NO和氧自由基的过度产生应该是各种感染中的普遍现象。诸如过氧亚硝酸盐之类的活性氮氧化物物种被生成…通过NO与O-ィイD3-(/)2-ィエD3的反应，在生物体系中得到了更多的应用。过氧亚硝酸盐及其生物学作用引起蛋白质和DNA的氨基酸残基的氧化和硝化、脂质过氧化和DNA裂解。事实上，我们目前的研究表明，过氧亚硝酸盐通过对宿主细胞和组织的非选择性损伤而在病毒发病机制中发挥关键作用。因此，了解NO和氧自由基在病毒感染中的作用，不仅有助于在分子水平上深入了解病毒的发病机制，而且有助于从分子水平上了解微生物感染中宿主与病原体的相互作用。此外，在NO诱导的各种病理效应中，NO在微生物病原体中的诱变潜力也是耐人寻味的。对于病毒突变，研究病毒的氧化应激和毒力之间可能存在的联系是很有意义的。在此背景下，我们最近的研究首次证实了高产量NO诱导的氧化应激加速了RNA病毒的突变。简单地说，通过使用含有基因突变标记基因的重组RNA病毒仙台病毒(一种负义单链RNA病毒)和iNOS基因敲除小鼠，我们获得了确凿和直接的证据，表明体内宿主(野生型小鼠)中过量产生的NO明显增加并加速了病毒突变率。这种加速突变的过程扩大了病原体变异的异质性，导致在选择压力下的快速进化。NO和OィイD3-(/)2ィエD3-因此过氧亚硝酸盐的产生在受感染的宿主中普遍存在。因此，这一发现对RNA病毒的总体进化具有重要意义，包括在体内快速产生耐药、免疫耐受和细胞嗜性改变的艾滋病毒突变体。较少

项目成果

H. Maeda and T. Akaike: "Nitric oxide and oxygen radicals in infection, inflammation, and cancer"Biochemistry (Moscow). 63. 854-865 (1998)

H. Maeda 和 T. Akaike：“感染、炎症和癌症中的一氧化氮和氧自由基”生物化学（莫斯科）。

赤池孝章: "特集/NO(一般化窒素)NOS阻害剤の臨床応用の可能性"循環器科. 44. 459-466 (1998)

Takaaki Akaike：“NO（广义氮）NOS 抑制剂的临床应用的特点/可能性”心脏病学系 44. 459-466 (1998)。

H. Sugiura, et al.: "Role of peroxynitrite in airway microvascular hyperpermeability during late allergic phase in guinea pigs"Am. J. Respir. Crit. Care Med.. 160. 663-671 (1999)

H. Sugiura 等人：“过氧亚硝酸盐在豚鼠过敏后期气道微血管通透性过高中的作用”Am。

T. Akaike, et al.: "Nanomolar quantification and identification of various nitrosothiols by high performance liquid chromatography coupled with flow reactors of metals and Griess reagent"J. Biochem.. 122. 459-466 (1997)

T. Akaike 等人：“通过高效液相色谱与金属和格里斯试剂的流动反应器联用对各种亚硝基硫醇进行纳摩尔定量和鉴定”J。

赤池孝章: "IV.炎症に関与する物質(室田誠逸、柏崎禎夫編)"(株)医薬ジャーナル社(東京). 11 (1997)

Takaaki Akaike：“IV. 与炎症有关的物质（由室田精一和柏崎贞雄编辑）”Iyaku Journal Co., Ltd.（东京）11（1997）。