MiR-206通过脂类代谢影响HCV感染和致病的机制研究

Hepatitis C virus (HCV) infection was the pathogenic cause of hepatitis C. Because of the complexity of its infected process and pathogenesis, the pathogenic mechanisms were not completely clear. It was reported that lipid metabolism participated in HCV infection, duplication, and pathogenesis, so HCV infection closely related with lipid metabolism. microRNA 206 (miR-206) was identified to be one important factor in lipid metabolism in liver, so we conferred that miR-206 might be affected HCV infection, duplication, and pathogenic process by lipid metabolism in liver. Firstly, we forecast target genes of miR-206 and validate them; secondly, mRNA and protein of miR-206 and its target genes were detected in cells with HCV infection. At same time, the ability of HCV infection and pathogenic process was detected after overexpressing and silencing miR-206. After comprehensive analysis, we construct the functional network of these genes to further clarify the role of miR-206 in HCV infected and pathogenic process by lipid metabolism. Our study will further improve the pathogenic mechanisms of HCV infection, and provide theoretical basis for newly biological drug for HCV treatment in clinic.

丙型肝炎病毒(HCV)感染是导致丙型肝炎的致病因素，由于其感染和致病过程复杂，目前尚未完全阐明具体的致病机制。已有研究表明HCV的感染、复制与致病过程均需要脂代谢蛋白的参与，与脂类代谢过程有着密切的关系；而microRNA 206 (miR-206) 近期被证明参与到肝组织脂类代谢过程中并发挥重要作用，因此推断miR-206可能通过脂类代谢途径影响HCV在宿主体内的感染、复制和致病的过程。本项目通过预测并确定miR-206调控的脂代谢相关基因，检测miR-206及其调控基因在HCV感染细胞中的表达改变情况；同时分析miR-206高表达或抑制后，对HCV感染和致病能力的影响，并确定其作用的脂代谢通路。通过综合分析，最终确定miR-206如何通过脂代谢途径影响HCV的感染和致病过程，构建功能网络图，进一步阐明HCV的感染和致病机制，并为HCV感染者临床治疗生物新药的研发奠定理论基础。

HCV感染导致慢性肝炎，且可发展为肝硬化和肝癌。虽然现阶段已有针对HCV感染的治疗药物，但是治愈率约为90%且HCV耐药毒株也逐年增多，HCV的防治仍然是一个较严重的公共卫生问题。MicroRNA被证明在细胞的生理过程和病例过程中均扮演着重要的角色。近期，MicroRNA-206（miR-206）被证实参与到脂类代谢的过程中。脂滴是HCV在细胞内增殖生命周期中具有极为重要的作用。而miR-206是否参与HCV感染及其增殖过程中并未知晓，因此本项目对此进行了研究。首先我们证明了miR-206转染HCV感染细胞中HCV RNA和蛋白水平的增殖均有较好的抑制作用，随后我们通过生物信息学方法预测了miR-206潜在的脂代谢通路的靶基因，将目的基因的3’UTR构建入pmirGLO质粒，通过双荧光素酶报告系统实验，证实miR-206对脂类合成基因ACC1具有直接抑制表达的作用。进一步的研究结果表明miR-206抑制了ACC1基因通路主要基因的RNA和蛋白水平的表达，而且miR-206成功的抑制了细胞内脂滴的产生。总之，我们通过本项目的支持，证明了miR-206通过抑制ACC1脂类合成通路抑制细胞内脂滴的形成，从而抑制HCV在细胞内的增殖。基于本研究结果，miR-206有望在未来开发为生物化学药物并应用于临床HCV感染的治疗。