基于TLRs/MyD88信号通路的银花平感颗粒抗甲型流感病毒免疫机制研究

Yinhuapinggan Granule was developed by our project team and was new drug of traditional Chinese medicines with independent intellectual property rights, which was composed of pungent-cool and pungent-warm drugs, and had the efficacy of clearing heat, freeing lung and relieving superficies. The formula of Yinhuapinggan Granule is precise and appropriate. The early efficacy and clinical trials showed that the Yinhuapinggan Granule has significant curative effect. But the mechanism of Yinhuapinggan Granule needs further research.. Objective：On basis of preliminary studies, and based on TLRs/MyD88 signaling pathway, to explore the influenza virus infection and Yinhuapinggan Granule on this signaling pathway, and to reveal the anti-influenza virus mechanism of Yinhuapinggan Granule.. Content: Guided by the theory of traditional Chinese medicine, based influenza a virus infection on TLRs/MyD88 signaling pathways, use a combination of the modern experimental technology and the multi-subject theories, adopted a method that combines the vitro test and the vivo test, focus from TLRs/MyD88 signaling pathway in major gene expression, protein expression and level of activity, such as TLR3, TLR7, MyD88, TRIF, IRF-3 and so on, to explore the regulation function and the anti-influenza virus mechanism of Yinhuapinggan Granule.. Meaning: Research the compatibility mechanism and treatment advantage of Yinhuapinggan Granule composed of pungent-cool and pungent-warm drugs, and disprove and explain the pathological biological basis of traditional Chinese medicine pathogenesis at the beginning of the flu, in order to expand the treatment thoughts, to guide clinical rational drug use, to improve the clinical efficacy, to establish a research method.

银花平感颗粒是本项目组研制具有自主知识产权的中药新药，本品辛凉与辛温合用，清热宣肺解表，组方精当，前期药效和临床试验表明其治疗流感初起具有显著疗效与作用，但尚需深入研究其作用机制。.目标：在前期研究基础上，基于TLRs/MyD88信号通路，探索流感病毒感染对该信号通路作用与银花平感颗粒调控作用，阐述本品抗流感病毒部分作用机制。.内容：以中医药理论为指导，基于甲型流感病毒感染对TLRs/MyD88信号通路作用，综合运用多学科理论与现代实验技术，采用体外与在体试验相结合，重点选择该信号通路主要基因、蛋白表达及活性水平指标(TLR3、TLR7、MyD88、TRIF、IRF-3等)，研究银花平感颗粒调控作用以阐述其抗流感作用机制。.意义：研究银花平感颗粒方辛凉与辛温配伍机制与治疗优势，反证并诠释流感初起中医病机学说的病理生物学基础，开拓治疗思路，指导临床合理用药，提高临床疗效，建立研究方法。

项目组在已有的研究基础上，本研究体内动物实验发现银花平感颗粒对甲型H1N1流感病毒感染小鼠具有治疗作用，其抗流感病毒作用机制可能与减轻流感病毒感染小鼠肺部损伤，提高流感病毒小鼠免疫功能和调控TLRs信号通路关键靶点的基因和蛋白表达有关。本研究体外实验研究发现银花平感颗粒对流感病毒H1N1的增殖有显著抑制作用，能减轻染毒MDCK细胞CPE，主要表现在直接灭活病毒、抗吸附作用以及治疗作用方面，其抗流感病毒感染的作用机制可能与其抑制TLR7/MyD88信号通路的激活，降低关键靶点mRNA表达，以及抑制IFN-β、IL-6的分泌有关；通过银花平感颗粒和三拗汤两方药体内体外结合对比研究对甲型H1N1流感病毒影响，探讨了不同方剂配伍对流感病毒的抑制作用。进一步验证了银花平感颗粒的组方科学性。开展了基于诱生IFN-α/β的TLRs/MyD88 信号通路研究银花平感颗粒抗A 型流感病毒免疫机制研究，取得了较好的研究成果。现已将相关研究成果整理撰写发表相关科研论文，正式发表10篇，其中SCI收录2篇（均带标注）；培养研究生2名；获省中医药科学技术一等奖、省部级二等奖。现已按原研究计划，在研究周期内完成了全部研究任务，为中药抗病毒研究提供先进、可行的技术和方法，为临床治疗提供思路和有效方药。