基于肠道菌群－宿主共代谢免疫交互网络调控效应研究哮宁口服液干预支气管哮喘的作用机制

Bronchial asthma is closely related to intestinal flora. By pre- clinical and experimental studies ,research group found that the curative effect of xiao ning liquid extract treatment bronchial asthma in children is accurate, and which has better control effect in intestinal flora disturbance, but its mechanism is not clear . The latest research progress of the co-metabolism and cross immunity network regulating effect of the host and intestinal flora provides a new pointcut for further revealling the mechanism of Traditional Chinese Medicine treatment asthma in children. The issue around the " xiao ning liquid extract plays its overall adjustment action by regulating the intestinal flora - host co-metabolism and cross immunity network effect " hypothesis , based on the preliminary studies , using the asthma model of OVA sensitization , integrating the PCR-DGGE molecular ecology , LC-MS metabolomics , flow cytometry (FCM) , immunohistochemistry and multivariate statistics combination strategy and multidisciplinary technology, to study the regulating effect of the Chinese medicine compound xiao ning liquid extract in the intestinal flora - host co-metabolism and cross immune network .Hoping from " intestinal flora - host interaction balanced " perspective , to further reveal the scientific connotation of overall regulatory mechanism of traditional Chinese medicine compound and the multi-layered, multiple- link complex action system of chinese medicine , which promoting theoretical reference and experimental evidence for the improvement of the clinical efficacy.

支气管哮喘与肠道菌群失调关系密切。课题组前期临床和实验研究发现，哮宁口服液治疗儿童支气管哮喘疗效确切，且对肠道菌群紊乱有较好的调控作用，但其作用机制还不十分清楚。肠道菌群与宿主具有共代谢免疫交互网络调控效应的最新突破性研究进展，为进一步深入揭示传统中医药治疗儿童哮喘的作用机制提供了崭新的切入点。本课题围绕“哮宁口服液通过调控肠道菌群－宿主共代谢免疫交互网络效应发挥其整体调节作用”的假说，在前期研究基础上，借助OVA致敏哮喘模型，整合运用PCR-DGGE分子生态学、LC-MS代谢组学、流式细胞术、免疫组化和多元统计学相结合的策略和多学科技术，研究中药复方哮宁口服液对肠道菌群与宿主共代谢免疫交互网络的调控效应，以期从“肠道菌群－宿主交互平衡”的角度，为进一步揭示传统中药复方整体调节的科学内涵及中医药多层次、多环节的复杂作用体系，促进临床疗效的提高提供理论参考和实验依据。

本项目在前期研究的基础上，借助OVA致敏哮喘模型，以院内制剂哮宁口服液为载体，整合运用Illumina高通量测序技术、LC-MS代谢组学、免疫组化和多元统计学相结合的策略和方法，研究哮宁口服液对肠道菌群与宿主共代谢免疫交互网络的调控效应，以期从“肠道菌群－宿主交互平衡”的角度，揭示哮宁口服液对支气管哮喘的作用机制。研究结果发现：①哮宁口服液对哮喘模型具有良好的治疗效应，在降低气道高反应性、减轻气道炎症等方面有确切疗效。②哮宁口服液对肠道微生物多样性指数、丰富度和均匀度具有较好的调控作用。乳酸菌属、葡萄球菌属含量变化差异较大，拟杆菌门相对丰度最高，其次为厚壁菌门，变形杆菌门，脱铁杆菌门，这可能是哮宁口服液影响哮喘治疗效应的关键功能菌。③哮宁口服液对哮喘模型小鼠血浆中的关键代谢物葡萄糖、天冬氨酸、谷氨酸、柠檬酸、3-磷酸甘油酸、硬脂酸、亮氨酸、异亮氨酸、丙氨酸存在良性调节作用，其可能为哮宁口服液影响哮喘的关键代谢物。④哮宁口服液对肠粘膜TOLL-2受体有较明显的正向调节作用。⑤哮宁口服液对哮喘模型菌群结构与宿主的血浆代谢表型、免疫间存在关联。中药复方哮宁口服液可能通过影响肠道菌群与宿主代谢免疫交互发挥其治疗效应。项目研究结果为进一步揭示中药复方治疗哮喘的分子作用机制，深化传统中药复方整体调节作用的现代科学内涵奠定了基础。