HEV感染致脑组织损伤的机制研究

项目名称： HEV感染致脑组织损伤的机制研究

项目编号： No.31472165

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 佘锐萍

作者单位： 中国农业大学

项目金额： 88万元

中文摘要： 本研究以长爪沙鼠和家兔为实验动物，应用Transwell 小室迁移试验和脑血管内皮及神经胶质细胞体外培养技术，同时结合激光共聚焦显微镜观察技术，运用超微病理学、免疫组化和免疫电镜技术、Wester Bloting和PCR技术，总体研究目标锁定于HEV侵入脑组织并造成脑神经细胞及间质组织损伤的发病过程及其机制，重点围绕HEV如何突破血脑屏障进入脑组织这个核心问题展开观察研究。在观察HEV感染过程中ORF2和ORF3的蛋白表达在脑组织细胞内的定性和定位的同时，系统观察HEV对脑神经细胞、血管内皮和胶质细胞结构和功能的损伤作用，并对脑血管内皮和胶质细胞膜上的炎症相关信号分子进行定性、定量的观测，从一个新的视角在细胞和分子水平上系统全面位地揭示HEV在脑神经细胞内的复制、装配和释放的机制及HEV感染引起脑组织损伤的机制，为深入揭示HEV的发病机理及HEV的防治提供新的科学依据和新的研究思路。

中文关键词： 致病机制；戊性肝炎病毒；脑损伤；血脑屏障；小胶质细胞

英文摘要： In this study, using Mongolian gerbils and rabbits as experimental animals, the application of Transwell migration assay and in vitro culture technique for cerebrovascular endothelial cells and glial cells , combined with confocal microscopy techniques, ultrastructural pathology, immunohistochemistry, immuno-electron microscopy, western bloting and PCR techniques, the overall goal of this study is focused on pathogenesis and mechanism of the HEV invading the brain nerve tissue and resulting in injury of brain nerve cells and interstitial tissue。This study will centre on answers to questions how HEV to break through the blood brain barrier into the brain tissue. The qualitatively and quantitatively protein expressions of ORF2 and ORF3 in brain tissue will be observed in HEV infection process, meanwhile, the damage effect on structure and function of brain nerve cells, cerebrovascular endothelial cells and glial cells of experimental animals infected with HEV will be studied systematically，as well as both qualitative and quantitative detection on signal molecules related to inflammation on the surface of cerebrovascular endothelial cells and glial cells will be carried. The provenge reveals systematically and comprehensively the mechanism of HEV replication, assembly and release in brain nerve cells and the pathogenesis of brain injury caused by hepatitis E virus infection at the cellular and molecular level. This research provides theoretical basis for further study on diagnosis and preventative treatment of HEV, which reveals the pathogenesis of HEV infection from a new perspective.

英文关键词： pathogenesis;hepatitis E virus;brain injury;blood-brain barrier;microglia