Hepatitis C virus NS5A and core proteins induce oxidative stress-mediated calcium signalling alterations in hepatocytes
Hepatitis C virus NS5A and core proteins induce oxidative stress-mediated calcium signalling alterations in hepatocytes
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DOI：
10.1016/j.jhep.2008.12.026
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发表时间：
2009-05-01
影响因子：
25.7
通讯作者：
Gonzalez-Gallego, Javier
中科院分区：
文献类型：
--
作者：
Dionisio, Natalia;Garcia-Mediavilla, Maria V.;Gonzalez-Gallego, Javier
背景/目的：丙型肝炎病毒 (HCV) 结构核心和非结构 NS5A 蛋白在肝细胞中诱导一系列细胞内事件，包括活性氧和氮 (ROS/RNS) 的升高。由于氧化应激与细胞内 Ca2+ 稳态的改变有关，我们的目的是研究这些蛋白质对人肝细胞来源的转染细胞中 Ca2+ 动员的影响，以及槲皮素治疗的保护作用。方法：通过荧光光度法测定 Ca2+ 动员和肌动蛋白重组。通过流式细胞术测定 ROS/RNS 的产生。结果：用 NS5A 和核心蛋白转染的细胞显示出 ROS/RNS 产生和静息胞质 Ca2+ 浓度增加，并降低进入储存的 Ca2+ 浓度。在转染细胞中，去氧肾上腺素引起的 Ca2+ 释放、质膜 Ca2+-ATP 酶的 Ca2+ 进入和排出均显着减少。在细胞因子激活的细胞中观察到类似的效果。核心蛋白和 NS5A 蛋白存在时，去氧肾上腺素诱发的肌动蛋白重组会减少。槲皮素可显着防止这些影响。在含复制子的细胞中观察到 Ca2+ 动员的改变和钙蛋白酶激活的增加。结论：NS5A 和核心蛋白可诱导人肝细胞来源的细胞中氧化应激介导的 Ca2+ 稳态改变，这可能是这两种蛋白在与 HCV 感染相关的肝脏疾病发病机制中的作用的基础。 (c) 2009 年欧洲肝脏研究协会。由 Elsevier B.V. 出版。保留所有权利。
Background/Aims: The hepatitis C virus (HCV) structural core and non-structural NS5A proteins induce in liver cells a series of intracellular events, including elevation of reactive oxygen and nitrogen species (ROS/RNS). Since oxidative stress is associated to altered intracellular Ca2+ homeostasis, we aimed to investigate the effect of these proteins on Ca2+ mobilization in human hepatocyte-derived transfected cells, and the protective effect of quercetin treatment. Methods: Ca2+ mobilization and actin reorganization were determined by spectrofluorimetry. Production of ROS/RNS was determined by flow cytometry.Results: Cells transfected with NS5A and core proteins showed enhanced ROS/RNS production and resting cytosolic Ca2+ concentration, and reduced Ca2+ concentration into the stores. Phenylephrine-evoked Ca2+ release, Ca2+ entry and extrusion by the plasma membrane Ca2+-ATPase were significantly reduced in transfected cells. Similar effects were observed in cytokine-activated cells. Phenylephrine-evoked actin reorganization was reduced in the presence of core and NS5A proteins. These effects were significantly prevented by quercetin. Altered Ca2+ mobilization and increased calpain activation were observed in replicon-containing cells.Conclusions: NS5A and core proteins induce oxidative stress-mediated Ca2+ homeostasis alterations in human hepatocyte-derived cells, which might underlie the effects of both proteins in the pathogenesis of liver disorders associated to HCV infection. (c) 2009 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.