肝细胞中高迁移率族蛋白1对病毒生活周期的影响

发布时间：2018-05-30 00:13

本文选题：丙型肝炎病毒 + HMGB1　；参考：《湖南大学》2013年硕士论文

【摘要】：丙型肝炎病毒（HCV）为单股正链RNA病毒，感染了全世界约1.7亿人，可导致慢性肝炎、肝硬化、肝癌等严重的肝脏疾病。联合使用聚乙二醇干扰素和利巴韦林是经典的丙肝治疗方法，但约50%的1型HCV病人对此耐药，研发丙肝的新疗法迫在眉睫，而对HCV生活周期的阐述则为抗病毒研究提供治疗的靶标。高迁移率族蛋白1（HMGB1）是一种非组蛋白核蛋白，调节染色质的结构和功能。在病毒感染宿主细胞时，HMGB1既能抑制登革热病毒和水泡性口炎病毒等病毒的复制，又能促进禽流感病毒的复制，本课题主要研究肝细胞中HMGB1对HCV生活周期的影响。我们运用siRNA和shRNA等RNA干扰手段，对肝癌细胞系Huh7、Huh7.5以及HLCZ01中的HMGB1进行基因沉默。在HCV感染Huh7.5和HLCZ01，以及NDV感染Huh7的实验中，沉默HMGB1导致细胞中的病毒含量降低，，过表达HMGB1则导致HCV在HLCZ01细胞中的含量升高，同时IFNβ的表达水平与细胞中病毒含量呈正相关性，这些结果预示着肝细胞中的HMGB1能够促进病毒复制，并且不是通过影响细胞的先天免疫反应实现的。免疫荧光结果显示，HCV的感染诱导了肝细胞中的HMGB1由胞核向胞质的转移。通过免疫共沉淀和RNA结合蛋白免疫共沉淀实验证实，HMGB1不与HCV的core、NS2、NS5A等病毒蛋白结合，而与HCV的负链RNA结合。这些实验数据显示，HCV感染肝细胞后，HMGB1从胞核转移至胞质，通过结合HCV负链RNA促进HCV的复制。我们的研究对于HMGB1在病毒感染中的功能，以及HCV的生活周期有了新的阐述，为抗病毒研究提供了一种新的靶标、思路和方法。
[Abstract]:Hepatitis C virus (HCV) is a single strand of positive chain RNA virus, infected with about 170 million people worldwide, and can lead to chronic hepatitis, cirrhosis, liver cancer and other serious liver diseases. The combination of peginterferon and Leigh Bhave Lin is a classic hepatitis C treatment, but about 50% of type 1 HCV patients are resistant to this, and the new therapy for hepatitis C is imminent. The elaboration of the HCV life cycle provides a target for the treatment of antiviral research. High mobility group protein 1 (HMGB1) is a non histone nucleoprotein that regulates the structure and function of chromatin. When the virus infects the host cell, HMGB1 can inhibit the replication of dengue virus and vesicular stomatitis virus and promote the avian influenza virus. In this project, we mainly studied the effect of HMGB1 on the life cycle of HCV in hepatocytes.
We use RNA interference such as siRNA and shRNA to silencing the HMGB1 in the cell lines of Huh7, Huh7.5 and HLCZ01. In HCV infected Huh7.5 and HLCZ01, and NDV infection Huh7, the silence caused the decrease of the virus content in the cells. There is a positive correlation between the expression level and the virus content in the cells. These results indicate that HMGB1 in the hepatocytes can promote the replication of the virus and is not achieved by affecting the innate immune response of the cells. The immunofluorescence results show that the infection of HCV induces the transfer of HMGB1 from the nucleus to the cytoplasm in the liver cells. By immunoprecipitation and RN A binding protein immunoprecipitation assay demonstrated that HMGB1 does not bind to HCV's core, NS2, NS5A and other viral proteins, and is associated with negative chain RNA of HCV. These experimental data show that HMGB1 is transferred from the nucleus to the cytoplasm after HCV infection of the liver cells, and promotes replication by combining HCV negative strand RNA. The new life cycle of HCV has provided a new target, train of thought and method for antiviral research.
【学位授予单位】：湖南大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R373.21

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