Nonstructural 3 protein of hepatitis C virus triggers an oxidative burst in human monocytes via activation of NADPH oxidase
Nonstructural 3 protein of hepatitis C virus triggers an oxidative burst in human monocytes via activation of NADPH oxidase
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DOI：
10.1074/jbc.m100698200
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发表时间：
2001-06-22
影响因子：
4.8
通讯作者：
Pipy, B
中科院分区：
文献类型：
--
作者：
Bureau, C;Bernad, J;Pipy, B
研究表明，氧化应激发生在慢性丙型肝炎中。从隔离的吞噬细胞和活化的驻留巨噬细胞释放活性氧（ROS）代表肝脏中氧化应激的主要组分。然而，很少有人知道单核细胞产生活性氧的能力，以响应丙型肝炎病毒的蛋白。在这项研究中，我们研究了几种丙型肝炎病毒蛋白刺激人单核细胞产生的ROS。将来自健康献血者的人单核细胞与重组病毒蛋白：Core、NS 3、NS 4和NS 5孵育。通过化学发光测量ROS产生。只有NS 3触发人单核细胞中的ROS产生。产生的活性氧主要是超氧阴离子。NS 3也诱导了一个快速和短暂的增加，细胞内钙离子浓度测量的视频数字显微镜技术。通过使用不同的代谢抑制剂，我们发现，ROS的产生需要钙内流，酪氨酸激酶，和应激活化蛋白激酶，p38。对p47（PHOX）磷酸化和易位的研究表明，NADPH氧化酶被激活并参与NS 3诱导的ROS产生。在第二个实验中，NS 3抑制由佛波醇12-肉豆蔻酸酯13-乙酸酯诱导的氧化爆发。这些结果表明，NS 3激活NADPH氧化酶并调节ROS的产生，这可能涉及丙型肝炎感染的自然史。
It has been shown that oxidative stress occurs in chronic hepatitis C. Release of reactive oxygen species (ROS) from sequestered phagocytes and activated resident macrophages represents the predominant component of oxidative stress in the liver. However, little is known about the ability of the monocyte to produce ROS in response to protein of hepatitis C virus. In this study, we investigated the ROS production in human monocytes stimulated by several viral proteins of hepatitis C virus. Human monocytes from healthy blood donors were incubated with recombinant viral protein: Core, NS3, NS4, and NS5. ROS production was measured by chemiluminescence. Only NS3 triggered ROS production in human monocytes. Generated ROS were mainly the anion superoxide. NS3 also induced a rapid and transient increase in intracellular calcium concentration measured by a video digital microscopy technique. By using different metabolic inhibitors, we showed that ROS production requires calcium influx, tyrosine kinases, and the stress-activated protein kinase, p38. The study of p47(PHOX) phosphorylation and translocation showed that NADPH oxidase was activated and involved in ROS production induced by NS3. In a second experiment, NS3 inhibited the oxidative burst induced by phorbol 12-myristate 13-acetate. These results indicate that NS3 activates NADPH oxidase and modulates ROS production, which may be involved in the natural history of hepatitis C infection.