Wnt5a在重症甲型H1N1流感发病机制中的作用研究

2009 pandemic H1N1 influenza's global prevalence deserves our attention. Patients with the influenza are mostly mild cases, while some are severe or critical that progress towards respiratory failure or even death. On the basis of preclinical, pathological, cytokines and animal model study about the H1N1 influenza, we point that the excessive inflammatory response is the main reason of organ damage or death induced by influenza. This project focuses on the mechanism of Wnt5a's inflammatory response in acute lung injury at the whole, organ, cell, protein and gene levels, through the study of the H1N1 influenza's animal model and detection of serum of patients. The main content of our work is to find the expression levels of Wnt5a and pro-inflammatory cytokines and the macrophage's infiltration degree, to study on Wnt5a's role in the H1N1 influenza with the animal model, and to further verify the hypothesis on acute lung injury induced by influenza though cytokines-mediated Wnt5a signaling and search for therapeutic target. This study intends to reveal the mechanism of severe influenza, and provide a theoretical basis and feasible option for the early diagnosis and prevention of severe influenza.

甲型H1N1流感全球流行值得关注。该型流感多为轻症，部分表现为重症或危重症，出现呼吸衰竭甚至死亡。在前期临床、病理、细胞因子和甲型H1N1流感动物模型研究基础上，我们提出过度炎症反应是流感患者脏器损伤乃至死亡的主要原因。本课题拟用甲型H1N1流感动物模型、患者血清和呼吸道分泌物，从整体、器官、细胞、蛋白和基因水平全面系统研究wnt5a在重症流感致急性肺损伤发病机制中的作用。主要研究内容为：Wnt5a、促炎细胞因子表达水平及巨噬细胞浸润程度；利用动物模型研究拮抗Wnt5a对重症甲型H1N1流感的干预效应，进一步通过验证Wnt5a通过炎症因子介导肺损伤的假说，寻找治疗靶点。本研究力求揭示Wnt5a在重症流感发病机制中的作用，为临床重症流感早期诊断和防控提供理论依据和可行方案。

流感对世界公共卫生已经构成了严重的威胁，甲型H1N1流感全球流行值得关注。该型流感多为轻症，部分表现为重症或危重症，出现呼吸衰竭甚至死亡。在前期临床、病理、细胞因子和甲型H1N1流感动物模型研究基础上，我们提出过度炎症反应是流感患者脏器损伤乃至死亡的主要原因。.本研究基于上述背景，应用ELISA方法检测重症甲流患者血清及肺泡灌洗液Wnt5a水平，采用Westblot以及分子生物学技术检测肺组织样本中Wnt通路相关蛋白表达情况，明确Wnt5a在重症流感发生发展中的作用；动物模型上，观察干预Wnt通路对重症甲型H1N1流感动物模型的影响，明确巨噬细胞、细胞因子和Wnt5a在肺损伤中的作用，为探讨重症甲型H1N1的发病机制提供依据。本课题发现重症流感患者Wnt5a水平升高，部分细胞因子水平升高；重症甲型H1N1流感动物模型Wnt5a水平升高，巨噬细胞肺浸润增加，部分炎症因子水平升高，Wnt通路下游分子β-catenin、DVL2、CyclinD1 表达增加；Wnt5a中和抗体对重症甲型H1N1流感的有保护效应，降低IL-6、RANTES水平，提示Wnt5a通过炎症因子介导肺损伤。同时根据目前流感患者的标本库，寻找与炎症因子相关的可以辅助判断疾病的生物标志物，为临床重症流感早期诊断和防控提供理论依据和可行方案。.通过基础与临床相关的研究，我们认为过度Wnt5a在重症流感发展中起到重要作用，调控Wnt5a可能作为重症甲型H1N1流感的治疗靶点。