VSIG4抑制MHV-3病毒感染诱发的暴发型肝衰竭的分子机制研究

项目名称： VSIG4抑制MHV-3病毒感染诱发的暴发型肝衰竭的分子机制研究

项目编号： No.81471949

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 杨承英

作者单位： 中国人民解放军第三军医大学

项目金额： 72万元

中文摘要： 巨噬细胞(Mφs)及其分泌的炎症因子可加剧鼠肝炎病毒株-3(MHV-3)诱发的暴发型肝衰竭（FH）。VSIG4是特异表达于Mφs表面的免疫分子。前期研究我们发现， MHV-3可刺激VSIG4缺陷Mφs产生过量活性氧（ROS）、导致NLRP3炎症小体活化及IL1β分泌,最终引发VSIG4 KO小鼠的快速死亡。但VSIG4调控ROS分泌的机制尚未知。本研究拟:使用p47phox KO小鼠感染MHV-3，明确ROS对MHV-3所致FH的影响；转染RAW264.7细胞表达VSIG4，利用qPCR，WB等分析VSIG4调节ROS分泌的分子机制；使用Co-IP、TAP-质谱技术鉴定VSIG4调控NAPDH氧化酶活性可能的新效应分子；在体给予ROS阻断剂或VSIG4激动型抗体，探讨基于VSIG4信号对FH的干预效果。本研究将阐明VSIG4抑制FH病理进程的作用机制，并为FH的临床免疫干预提供新策略。

中文关键词： MHV-3；VSIG4；巨噬细胞；ROS；暴发型肝衰竭

英文摘要： Macrophages (Mφs) and their secreted proinflammatory factor can aggravate the pathogenesis of fulminant hepatic failure (FH) induced by murine hepatitis virus 3(MHV-3), which can be delayed by inhibiting the inflammation caused by macrophages. V-set and Ig domain-containing 4(VSIG4) is a co-inhibitory molecule specifically expressed on the surface of tissue macrophages, such as peritoneal macrophages and kupffer cells. In our previous study, we found that VSIG4 deficiency exacerbates MHV-3 induced liver injury and mortality and which is depend on IL-1β. VSIG4 deficiency macrophages produce more reactive oxygen species (ROS) which increase NLRP3 inflammasom and secrete more IL-1β than WT. These results indicated that VSIG4 maybe contribute to delay the pathogenesis of FH by down-regulating the expression of ROS. However，the molecular mechanism about VSIG4 regulate ROS is not yet clear. In this study， we want to infect p47phox knockout mice with MHV-3 to analyze the role of ROS in FH induced by MHV-3, transfect RAW264.7 cells with VSIG4 expression vector to study the mechanism about VSIG4 regulate ROS using qPCR, Western Blot and EMSA, screen the proteins interacting with VSIG4 using TAP-chip and co-IP to identify the new effector molecules of VSIG4 targeting. In addition, the probably therapeutic effect of VSIG4 signal pathway should be evaluated by using VSIG4 agonist antibodies or ROS inhibitors treatment in vivo. This study will clarify the mechanism of VSIG4 involved in regulating the process of FH, which provide a new strategy to the therapy of FH.

英文关键词： MHV-3;VSIG4;macrophage;ROS;fulminant hepatic failure