中药射干抗气道变应性炎症多靶标作用机制及配伍关系研究

Airway allergic inflammation was one of the major causes on cough due to virus cold, which led to the poor or ineffective therapeutic outcome by antihistamine H1 receptor antagonists, central antitussive, and antibiotics et al. Airway allergic inflammation belonged to the exogenous cough in traditional Chinese medicine, which symptoms was inner invasion superficial pathogens. With the therapeutic principle of multiple components and targets, good results have been achieved. Previous research suggested that the different isoflavones in Rhizoma Belamcandae had good anti-inflammatory, and antitussive effects respectively. Belamcanda chinensis extract consist of isoflavones with broad-spectrum anti-virus capabilities can significantly inhibit metabolic pathway of arachidonic acid lipoxygenase and cyclooxygenase, which indicated whole regulation and multi-target superiority of the traditional Chinese medicine in treatment. This project is to study the extracts and monomer compounds from Belamcanda chinensis on the arachidonic acid metabolic pathways, and signal transduction pathway, et al of the inflammation of guinea pig cough model caused by syncytial virus. The mechanisms of airway allergic inflammation caused by viral influenza symptoms of asthma were confirmed on tissular, cellular and molecular level. Based on dose-effect relationship between monomer compounds from Belamcanda chinensis and therapeutic effects, correlation analysis and regression analysis were applied to build the multi-level networks about constituent and effect target and explore the compatibility of isoflavones in Rhizoma Belamcandae.

气道变应性炎症是病毒性感冒后咳嗽的主要原因，抗组织胺H1受体拮抗剂、中枢镇咳药、抗生素等化学药品治疗效果欠佳或无效。该病属于中医学"外感咳嗽病"范畴，其中医症候特点为表邪未尽，多年来通过多成分、多靶标治疗原则，取得良好效果。前期研究发现，射干不同异黄酮类化合物分别具有良好的抗炎、止咳等多靶标作用；由它们组成的射干提取物具有抗病毒谱广，显著抑制花生四烯酸脂氧合酶及环氧合酶代谢途径效果。体现了中药多组分整体调理、多靶标协同治疗该病的优势。本项目拟通过射干提取物及其不同活性单体化合物对豚鼠合胞病毒感染性咳嗽模型气道炎症的花生四烯酸代谢途径、细胞信号传导途径等多方面影响研究，从组织、细胞、分子水平上确定其抗病毒性感冒后咳嗽所引起气道变应性炎症的作用机理。同时根据射干提取物中单体化合物与疗效之间的量-效关系，利用相关分析及回归分析，构建成分-成分-靶点的多层次网络，明确不同异黄酮类化合物之间配伍关系

摘 要.气道炎症可由细菌或病毒介导，气道上皮损伤，炎性细胞活化并浸润于气道，导致大量细胞因子和炎性介质产生和释放，气道神经及神经肽的表型和含量均发生改变，Sp及其受体为主的神经肽或者VR1等神经递质的释放增加，使咳嗽反射感受器敏感性增加，并使以磷脂形式存在于细胞膜上的花生四烯酸（AA）在PLA2和PLC催化下，从磷脂池中释放出来，转变为具有生物活性的代谢产物，发生AA的炎症级联代谢。.围绕气道炎症发病机制，探讨射干异黄酮单体成分及配伍对合胞病毒感染豚鼠咳嗽模型变应性气道炎症的（1）咳嗽次数（2）炎性细胞（3）细胞因子（4）肺组织形态（5）SP物质含量（6）Sp受体NK1、辣椒素受体VRl蛋白表达（7）受体NK1、VR1 mRNA表达（8）花生四烯酸代谢产物影响，从整体、器官、细胞及分子不同药理层次上揭示射干异黄酮化合物抗变应性气道炎症的作用机理；并通过相关分析和回归分析，明确各单体成分作用途径和机制；确定成分间最佳配伍剂量。. 结果：明确了射干抗气道炎症多中心、多靶点的作用机制，不同的作用途径及成分间最佳配伍：（1）鸢尾黄素、白射干素及野鸢苷可显著抑制豚鼠咳嗽；（2）除射干苷外，另外5种成分可分别降低不同种类的炎性细胞计数；（3）鸢尾黄素、野鸢尾黄素及白射干素可显著下调细胞因子IL-4/IFN-γ比值；（4）6个异黄酮成分都可减轻肺组织炎性浸润、下调模型肺组织SP含量、下调SP受体NK1、辣椒素受体VRl蛋白表达、下调受体NK1和VR1蛋白的mRNA表达；（5）同一化学成分在不同样本（血清、肺泡灌洗液、肺组织）中影响花生四烯酸代谢的途径不同，鸢尾黄素、次野鸢尾黄素、射干苷及野鸢尾苷影响主要代谢途径COX、P450及LOX，而野鸢尾黄素及白射干素只影响COX及LOX途径。(6) 不同配伍组对气道炎症的作用途径及强度与单一成分相比有所不同，经均匀设计软件处理，得到了射干异黄酮苷元抗气道炎症的最佳配伍比例为：鸢尾黄素-野鸢尾黄素-次野鸢尾黄素-白射干素（30:45:3:10）。