多种病毒可借助细胞微丝骨架的形态改变及异常重组来完成其侵入及在细胞内的运输过程。课题组前期研究发现，猪血凝性脑脊髓炎病毒（PHEV）入侵神经细胞触发了微丝主要成分纤维形肌动蛋白（F-actin）形态结构的改变，进而促进了病毒的增殖，但尚不清楚F-actin在该病毒侵入神经细胞的作用及其机制。鉴于此，本项目拟在建立细胞感染模型基础上，利用荧光染色、侵入动力学和阻断动力学实验，分析F-actin在PHEV入侵神经细胞过程中的变化特点及其对病毒复制的影响；利用蛋白活性检测、表达显性抑制突变体、抑制剂阻断等方法，明确PHEV入侵过程中F-actin发挥作用所依赖的分子基础；利用单颗粒追踪、边界-边界分布法及相关信号通路激活检测等方法，解析F-actin在该病毒胞内运输中的作用。预期成果不仅可阐明F-actin介导PHEV入侵神经细胞及胞内运输的作用机制，而且可为寻找新的抗病毒靶点药物提供理论依据。

Viruses complete invasion and intracellular trafficking through morphological changes and abnormal recombination of microfilament cytoskeleton. Based on our previous studies, porcine hemagglutinating encephalomyelitis virus (PHEV) invasion of nerve cells triggers the morphological structure changes of F-actin, which is the main component of microfilaments, and then promotes the virus proliferation. However, the dynamic regulation mechanisms of F-actin during PHEV invasion of nerve cells are unclear. In this study, the role of F-actin in PHEV invasion of nerve cells will be analyzed by fluorescence staining, invasion kinetics and blocking kinetics based on PHEV infected neural cell model. Then, molecular mechanisms of F-actin in PHEV invaded nerve cells will be studied through detecting protein activity, dominant negative mutants expression, inhibitor blocking and so on. Furthermore, the role of F-actin in PHEV intracellular trafficking will be analyzed by single-particle tracking, border-boundary distribution and related signaling pathway activation assays. The expected results will not only elucidate the molecular mechanism by which F-actin mediates PHEV invasion of nerve cells and intracellular trafficking, but also provide a theoretical basis for finding novel antiviral targeted drugs.