Zaragoza C, Ocampo C J, Saura M, McMillan A, Lowenstein C J
Division of Cardiology, Department of Medicine, School of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
J Clin Invest. 1997 Oct 1;100(7):1760-7. doi: 10.1172/JCI119702.
Nitric oxide is a radical molecule with antibacterial, -parasitic, and -viral properties. We investigated the mechanism of NO inhibition of Coxsackievirus B3 (CVB3) replication in vitro by determining the effect of NO upon a single replicative cycle of CVB3 grown in HeLa cells. Transfection of inducible NO synthase cDNA into HeLa cells reduces the number of viral particles produced during a single cycle of growth. Similarly, a noncytotoxic concentration of the NO donor S-nitroso-amino-penicillamine reduces the number of viral particles in a dose-dependent manner. To explore the mechanisms by which NO exerts its antiviral effect, we assayed the attachment, replication, and translation steps of the CVB3 life cycle. NO does not affect the attachment of CVB3 to HeLa cells. However, NO inhibits CVB3 RNA synthesis, as shown by a [3H]uridine incorporation assay, reverse transcription-PCR, and Northern analysis. In addition, NO inhibits CVB3 protein synthesis, as shown by [35S]methionine protein labeling and Western blot analysis of infected cells. Thus, NO inhibits CVB3 replication in part by inhibiting viral RNA synthesis by an unknown mechanism.
一氧化氮是一种具有抗菌、抗寄生虫和抗病毒特性的自由基分子。我们通过测定一氧化氮对在HeLa细胞中生长的柯萨奇病毒B3（CVB3）单个复制周期的影响，研究了一氧化氮在体外抑制CVB3复制的机制。将诱导型一氧化氮合酶cDNA转染到HeLa细胞中可减少单个生长周期内产生的病毒颗粒数量。同样，一氧化氮供体S-亚硝基-氨基青霉胺的非细胞毒性浓度以剂量依赖方式减少病毒颗粒数量。为了探究一氧化氮发挥其抗病毒作用的机制，我们检测了CVB3生命周期的附着、复制和翻译步骤。一氧化氮不影响CVB3与HeLa细胞的附着。然而，如[3H]尿苷掺入试验、逆转录-PCR和Northern分析所示，一氧化氮抑制CVB3 RNA合成。此外，如[35S]甲硫氨酸蛋白标记和对感染细胞的Western印迹分析所示，一氧化氮抑制CVB3蛋白合成。因此，一氧化氮部分通过未知机制抑制病毒RNA合成来抑制CVB3复制。
