Korenaga Masaaki, Wang Ting, Li Yanchun, Showalter Lori A, Chan Tehsheng, Sun Jiaren, Weinman Steven A
Center for Hepatitis Research, Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, 77555, USA.
J Biol Chem. 2005 Nov 11;280(45):37481-8. doi: 10.1074/jbc.M506412200. Epub 2005 Sep 8.
Hepatitis C infection causes a state of chronic oxidative stress, which may contribute to fibrosis and carcinogenesis in the liver. Previous studies have shown that expression of the HCV core protein in hepatoma cells depolarized mitochondria and increased reactive oxygen species (ROS) production, but the mechanisms of these effects are unknown. In this study we examined the properties of liver mitochondria from transgenic mice expressing HCV core protein, and from normal liver mitochondria incubated with recombinant core protein. Liver mitochondria from transgenic mice expressing the HCV proteins core, E1 and E2 demonstrated oxidation of the glutathione pool and a decrease in NADPH content. In addition, there was reduced activity of electron transport complex I, and increased ROS production from complex I substrates. There were no abnormalities observed in complex II or complex III function. Incubation of control mitochondria in vitro with recombinant core protein also caused glutathione oxidation, selective complex I inhibition, and increased ROS production. Proteinase K digestion of either transgenic mitochondria or control mitochondria incubated with core protein showed that core protein associates strongly with mitochondria, remains associated with the outer membrane, and is not taken up across the outer membrane. Core protein also increased Ca(2+) uptake into isolated mitochondria. These results suggest that interaction of core protein with mitochondria and subsequent oxidation of the glutathione pool and complex I inhibition may be an important cause of the oxidative stress seen in chronic hepatitis C.
丙型肝炎病毒感染会引发慢性氧化应激状态,这可能会促使肝脏发生纤维化和癌变。先前的研究表明,丙型肝炎病毒核心蛋白在肝癌细胞中的表达会使线粒体去极化,并增加活性氧(ROS)的产生,但这些效应的机制尚不清楚。在本研究中,我们检测了表达丙型肝炎病毒核心蛋白的转基因小鼠肝脏线粒体以及与重组核心蛋白孵育的正常肝脏线粒体的特性。表达丙型肝炎病毒核心蛋白、E1和E2的转基因小鼠肝脏线粒体表现出谷胱甘肽池的氧化和NADPH含量的降低。此外,电子传递复合体I的活性降低,复合体I底物产生的ROS增加。复合体II或复合体III的功能未观察到异常。用重组核心蛋白体外孵育对照线粒体也会导致谷胱甘肽氧化、选择性复合体I抑制以及ROS产生增加。用蛋白酶K消化与核心蛋白孵育的转基因线粒体或对照线粒体表明,核心蛋白与线粒体紧密结合,仍与外膜结合,且不会穿过外膜被摄取。核心蛋白还增加了分离线粒体对Ca(2+)的摄取。这些结果表明,核心蛋白与线粒体的相互作用以及随后谷胱甘肽池的氧化和复合体I的抑制可能是丙型肝炎慢性期所见氧化应激的重要原因。