从TLR2/4-MyD88信号通路研究流行性感冒“肺病及肠”的分子机制与麻杏石甘汤的调控作用

30.9% of influenza patients in addition to the performance of fever, headache and other systemic symptoms and cough, nasal congestion and other respiratory symptoms, but also accompanied by diarrhea, nausea, vomiting and other gastrointestinal symptoms. Although the disease "lung disease affected intestinal" performance has been concerned, but its molecular mechanism is not reported. Based on the theory of "lung and large intestine", the model was based on the "mucosal immune system". The model of influenza virus pneumonia was used to study the effects of respiratory and gastrointestinal function, lung tissue and intestinal tissue pathological damage and viral load To study the pathological mechanism of "lung disease and intestine" and the intervention effect of Maxing Shigan Decoction (Influenza prevention and control)；The expression of TLR2/4-MyD88 signaling pathway in inflammatory mediators was studied by real-time quantitative PCR, Western blot and laser confocal microscopy. The expression ofTLR2/4-MyD88 signaling pathway was induced by gene and protein expression. The main "element" changes and the regulation of the role of Maxing Shigan Decoction; Finally, with the influenza virus pneumonia model, from the passage of signal changes and respiratory function, gastrointestinal function, lung tissue and intestinal tissue pathological damage of the study of its "lung disease and intestinal" molecular mechanism and the mechanism of regulation, Lung disease and intestinal "scientific connotation, to expand the clinical application of Maxing Shigan Decoction

30.9%的流感患者除表现发热、头痛等全身症状和咳嗽、鼻塞等呼吸道症状外，还伴有腹泻、恶心、呕吐等胃肠炎症状。尽管该病“肺病及肠”之表现已被关注，但其分子机制尚无报道。本项目以“肺与大肠相表里”理论为依据，以“黏膜免疫系统”为依托，先用流感病毒肺部感染模型,从呼吸功能和胃肠功能、肺组织和肠组织病理损伤及病毒载量研究其“肺病及肠”病理机制与麻杏石甘汤（流感防治中倍受重视）干预效应；再用流感病毒刺激肺、肠巨噬细胞，以实时定量PCR、Western blot、激光共聚焦显微技术等，从基因与蛋白表达水平、活化与功能状况研究诱生炎症因子的TLR2/4-MyD88通路主要“元件”的变化与麻杏石甘汤调控作用；最后用流感病毒肺部感染模型，从信号通路改变与呼吸功能、胃肠功能、肺组织和肠组织病理损伤的相关性研究其“肺病及肠”分子机制与麻杏石甘汤调控机制，注释流感“肺病及肠”科学内涵，拓展麻杏石甘汤的临床应用。

胃肠炎型流感是流感的常见临床类型。尽管其临床表现已被关注，但其机制有待研究。本项目以“肺与大肠相表里”理论为依据，开展以下研究：①流感病毒肺部感染模型“肺病及肠”病理机制与麻杏石甘汤干预效应的研究；②流感病毒感染对肺、肠巨噬细胞TLR2/4介导的炎症因子分泌水平的影响及麻杏石甘汤调控作用的研究；③流感病毒感染对肺、肠巨噬细胞（组织）TLR2/4介导的趋化因子蛋白表达水平的影响及麻杏石甘汤调控作用的研究；④流感病毒肺部感染模型肺、肠组织TLR2/4-MyD88通路主要“元件”表达与活化水平改变与“肺病及肠”病理损伤的相关性及麻杏石甘汤干预效应的研究。结果是：①流感病毒肺部感染模型肺组织炎症损伤的同时，可伴有结肠组织的炎症、充血等病理变化，其机制可能与IL-6、MCP-1表达水平变化有关，麻杏石甘汤对模型小鼠IL-6、MCP-1血清含量水平及肺组织蛋白表达水平有调节作用。②流感病毒能促进TLR2/4介导的肺巨噬细胞 IL-1、 IL-8、TNF-α和肠巨噬细胞IL-8、TNF-β分泌，麻杏石甘汤含药血清对其有调节作用；流感病毒能促进肺、肠巨噬细胞TLR2、TLR4蛋白表达，麻杏石甘汤含药血清对肺巨噬细胞TLR2、TLR4及肠巨噬细胞TLR4蛋白表达水平有调节作用。③流感病毒能促进TLR2/4介导的肺巨噬细胞CCL3、CCL25与肠巨噬细胞CCL2分泌，麻杏石甘汤含药血清对其分泌水平有调节作用。麻杏石甘汤能明显调节流感病毒感染小鼠肺组织、肠组织CCL2、CCL3、CCL25蛋白表达水平。④给药3天时，麻杏石甘汤能明显调节流感病毒感染小鼠肺组织、肠组织TLR2、TLR4、MyD88、NF-κB蛋白表达水平。流感病毒感染小鼠肺组织炎症与TLR2/4-MyD88通路主要“元件”蛋白表达水平改变的密切相关。其结果丰富了流感“肺病及肠”科学内涵，为拓展麻杏石甘汤的临床应用提供实验支撑。