基于线粒体Mfn2/MAVS研究应激负荷诱导流感病毒易感性及清热消炎宁的干预作用

The anti-influenza mechanism of traditional Chinese medicine (TCM) is related to the regulation on viral susceptibility. However, the underlying biological mechanism is still unclear. Previous study demonstrated that restraint stress could elevate the level of stress-responsive corticosterone (CORT), causing increased viral susceptibility in mice. The mechanism behind was proposed to be related to the down-regulation of IFN-β through the Mfn2/MAVS pathway, but this hypothesis still needs to be proven. As Qingre Xiaoyan Ning could lower H1N1 susceptibility (ZL 2012102340) and up-regulate IFN-β level in restraint stressed mice, the current project aimed to study the influence of CORT on viral susceptibility and the anti-influenza mechanism of TCM, targeting Mfn2/MAVS signaling pathway using Qingre Xiaoyan Ning and restraint stressed mice model. This project also intended to use knockout-mice and RNAi technology in restraint stress mice model and CORT-loaded cell model to confirm the importance of Mfn2/MAVS in viral susceptibility. The pathomorphology of lungs, gene and protein expression of NP and Mfn2/MAVS/IRF3/IFN pathway will also be detected. This project also aims to reveal the biological mechanism of influenza susceptibility, to develop common technology for the evaluation of anti-influenza effect of TCM, and to elucidate the scientific connotation of anti-influenza effect of TCM.

中药防治流感作用与调节机体病毒易感性相关，但生物学机制并不清楚。前期研究提示拘束应激增加应激激素皮质酮（CORT）水平、提高小鼠对流感病毒的易感性，机制可能与影响线粒体Mfn2/MAVS通路、下调IFN-β水平相关，但需进一步验证。鉴于中药清热消炎宁能有效降低应激诱导的流感病毒易感性（ZL 2012102340），且能上调肺组织IFN-β水平。因此，本项目拟通过清热消炎宁给药病毒负荷应激小鼠，以Mfn2/MAVS信号通路为靶点，研究CORT对易感性的影响及中药抗病毒的作用机理。拟在拘束应激动物和CORT负荷细胞等模型上，采用基因敲除小鼠及RNAi技术确认Mfn2/MAVS在病毒易感性中的作用，检测肺组织病理变化、病毒核蛋白及RNA水平，检测Mfn2、MAVS、IRF3、IFN-β表达水平。旨在揭示流感病毒易感性的生物学机制，建立中药抗病毒药效评价的共性技术，阐明中药抗病毒作用的科学内涵。

中药防治流感作用与调节机体病毒易感性相关，但生物学机制并不清楚。前期研究提示拘束应激增加应激激素皮质酮（CORT）水平、提高小鼠对流感病毒的易感性，机制可能与影响线粒体Mfn2/MAVS通路、下调IFN-β水平相关，但需进一步验证。鉴于中药清热消炎宁能有效降低应激诱导的流感病毒易感性（ZL 2012102340），且能上调肺组织IFN-β水平。因此，本项目拟通过清热消炎宁给药病毒负荷应激小鼠，以Mfn2/MAVS信号通路为靶点，研究CORT对易感性的影响及中药抗病毒的作用机理。拟在拘束应激动物和CORT负荷细胞等模型上，采用基因敲除小鼠及RNAi技术确认Mfn2/MAVS在病毒易感性中的作用，检测肺组织病理变化、病毒核蛋白及RNA水平，检测Mfn2、MAVS、IRF3、IFN-β表达水平。旨在揭示流感病毒易感性的生物学机制，建立中药抗病毒药效评价的共性技术，阐明中药.抗病毒作用的科学内涵。