甘露消毒丹及其挥发油对IV感染小鼠粘膜免疫及RIG-Ⅰ/NF-κB信号通路作用机制的研究

Influenza virus (IV) is one of most serious virus threatening human health. Its pathogenic mechanism is that the invasion and potentiation mediated by mucosal immunity and specific signal pathway which induced the apoptosis and pulmonary injury. IV infection in a pollution haze is characterized as “damp-toxin” manifestation. Based on the pathogenic character, we innovate a hypothesis that treatment of IV infected pneumonia should be dealing with “damp-toxin” and “lung stuffiness”. Ganlu xiaolu micropills are best known as major formula treating damp-warm causing epidemic, functioned as repelling foulness, clearing heat, expelling damp and detoxifying with remarkbale clinical effect. The whole recipe and its essential oil are divided into two administration routes as oral and nasal inhalation. Begaining with active substance research, we detect index components by HPLC to find the best procedure. Study is a factorial design. Balb/c mice infected by IV are divided into with pollution haze group and without pollution haze group to observe mucosal immune response and dynamic change of components through RIG-Ⅰ/NF-κB dependent-signal pathway with techniques of Western-blot, RT-PCR and so on. For providing research and theory basis, our study attempt to explore the content of “damp-toxin” through modern biological indicators, to compare effectiveness of ganlu xiaolu micropills and to discuss the mechanism of integrated regulation how ganlu xiaolu micropills improve the immune pulmonary injury caused by IV.

流感病毒(IV)是对人类健康威胁最严重的病毒之一，主要是通过粘膜免疫、信号通路介导入侵、增毒，损伤细胞，诱导凋亡而导致肺损伤。根据雾霾条件下IV感染具有“湿毒”特征，创新性提出从“湿毒”、“肺痹”论治IV肺炎。甘露消毒丹为治湿温时疫之主方，芳香辟秽、清热祛湿、解毒化浊，临床疗效确切。全方及挥发油分别口服和滴鼻，“一方两用”，采用HPLC检测指标性成分确定最佳提取工艺，以其有效物质群为切入点，初步探寻物质基础及作用靶器官。通过析因设计，以Balb/c小鼠肺炎模型（有无雾霾环境）为对象，采用Western-blot、RT-PCR等技术方法，观察粘膜免疫、RIG-Ⅰ/NF-κB通路“元件”动态变化，阐明“湿毒”、“肺痹”现代生物指标内涵，探讨甘露消毒丹药能效果差异性，揭示改善IV免疫炎性损伤的整合调节机制，为“湿毒”论治IV感染临床应用提供实验依据与基础。

流感病毒(IV)是对人类健康威胁最严重的病毒之一，根据雾霾条件下IV感染具有“湿毒”特征，甘露消毒丹为治湿温时疫之主方，芳香辟秽、清热祛湿、解毒化浊，临床疗效确切。其全方及挥发油可分别口服和滴鼻，“一方两用”。本课题组创新性提出“湿毒”、“肺痹”并与IV肺炎的生物学指标关联，研究甘露消毒丹及其挥发油干预靶器官、靶向效应机制。以绿原酸、黄芩苷、连翘苷的含量作为工艺评价指标，采用正交设计筛选甘露消毒丹最佳提取工艺，采用HPLC检测甘露消毒丹中活性成分，对其进行质量控制研究。通过多层面验证IV滴鼻造模，并比较有无雾霾条件（PM2.5），发现肺部炎症、炎症因子、免疫功能等指标变化呈一致性，RIG-I/NF-κB信号通路亦被激活，模拟雾霾条件+IV滴鼻组呈现重症肺炎改变，且其指标表达更为明显。本研究从不同指标、不同角度及不同水平探讨甘露消毒丹及其挥发油抗IV感染的调控机制及优势靶点。甘露消毒丹可降低IV感染后肺部炎症，降低NS1蛋白的表达，减少促炎因子的释放，抑制细胞凋亡，调控RIG-I/NF-κB信号通路，同时能诱生干扰素，调节免疫，其滴鼻灌胃合用较单一给药有优势。甘露消毒丹能够调节粘膜免疫、维持免疫稳态与活化之间的平衡，可升高sIgA含量，降低炎症因子，挥发油滴鼻较醇提物灌胃作用明显，且起效迅速，挥发油滴鼻给药方式具有一定的优势。