麻黄-杏仁药对减轻气道损伤的配伍机理研究

Asthma is one of the four chronic and stubborn disease, Glucocorticoid is still the preferred treatment of asthma. Recent studies showed that the severity of asthma is proportional to the scope of airway epithelium damage and the degree of deficiency repair functions mediated by EGFR, Therefore, the drug to develop epithelial protective effect is one of the new research direction in the future. Traditional Chinese medicine has a long history of treating asthma, the curative effect is reliable. The most used medicine of treating asthma decoction is ephedra and almond. From interpreting the relationship between compatibility rules between components and effect is an important field of research due to the compound, Couplet medicines is the main object of study because it use a small number of medicine but strong pharmacological action. Previous study found that: Maxingshigan Decoction significantly reduced airway injury, the main ingredients are ephedra, almond (optimal dose ratio is 4:5), but its scientific connotation is not clear. In this study, couplet medicines of ephedra and almond was studied as the object, pharmacological methods of the bowel absorption liquid containing medicine composition is used. Technology of RRLC-QTOF/MS was used to analysis the chemical changes in compatibility. Technology of HCS, RT-PCR and Pearson analysis were used to investigate the correlation between anti airway injury and the key components of the EGFR/PI3K pathway multi-level. By using principal component analysis, to preliminary explane the scientific connotation of the regular pattern among compatibility, ingredients and effect. Through the study, we want to deepen the understanding of couplet medicines of ephedra and almond, provide demonstration and new ideas for the research on the compatibility, promote the modern interpretation of Chinese compound.

哮喘是四大顽症之一，糖皮质激素仍然是疗喘的首选。新近研究表明，哮喘的严重程度与气道上皮的损伤范围、EGFR介导的修复功能缺失程度成正比，因此，以开发具有上皮保护作用的药物是今后研究的新方向之一。中医药治喘历史悠久、疗效可靠，疗喘方运用最多的药味是麻黄、杏仁。从成分和效应之间的关系阐释配伍规律是复方研究的重要领域，药对由于药简力宏是主要研究对象。前期研究发现：麻杏石甘汤可明显减轻气道损伤，主要药味是麻黄、杏仁（最佳剂量配比4：5），但其内涵尚不清楚。本研究以麻黄-杏仁药对为对象，应用含药肠吸收液方法，采用RRLC-QTOF/MS分析配伍的成分变化；HCS、RT-PCR技术和Pearson分析，多层次探讨其抗气道损伤与EGFR/PI3K通路关键元件的相关性；并经主成分分析，初步阐释其"配伍-成分-效应"之间的规律。深化麻黄-杏仁药对的认识，为配伍研究提供示范和新的思路，促进复方的现代阐释。

哮喘是四大顽症之一，临床平喘首选是不良反应多的糖皮质激素；研究表明，哮喘的严重程度与气道上皮的损伤范围、EGFR介导的修复功能缺失程度成正比，因此，开发上皮保护药物是今后研究的新方向之一。阐明物质基础、作用机理及其关系是复方研究的重要领域，药对因药简力宏是主要对象；中医各疗喘方运用最多的药味是麻黄、杏仁。.本研究以麻黄-杏仁药对为对象，采用RRLC-QTOF/MS技术建立其定性定量分析方法，获取其不同配比质谱图后经鉴别和分析，并与已有化合物数据库比对，找出其主要差异性有效成分。应用含药肠吸收液方法，从整体、器官和细胞等水平、多层次探讨药对疗喘与抗气道损伤、调节EGFR/PI3K通路关键元件表达的相关性，揭示其疗喘新靶点。结合质谱和药效学结果，经主成分分析，初步阐释其“配伍-成分-效应”之间的规律及麻黄、杏仁“相须为用”的内涵。.结合Scifinder等数据库及文献报道，建立了麻黄、杏仁药味共57种化学成分的目标数据库及8种成分的有效化合物库；建立了药对含药肠吸收液的UPLC-Q-TOF-MS/MS分析方法，对不同配比质谱图进行鉴别、分析并与目标数据库匹配，确定出10种差异性化合物，再与有效化合物库匹配，基本厘清了药对的主要差异性有效成分：麻黄碱、伪麻黄碱、甲基麻黄碱、苦杏仁苷。从整体、器官和细胞的药效学研究表明，药对能延长引喘潜伏期有良好的平喘作用，该效应与其调节上皮细胞EGFR基因与蛋白的表达、减少炎症因子释放，从而减轻气道上皮细胞损伤密切相关；经最佳配比分析发现，麻黄：杏仁≥1：1的配比组对大多数效应指标的作用强于麻黄：杏仁<1：1的配比组。主成分分析表明，麻黄成分对各效应指标的影响明显强于杏仁成分，故麻黄为君药；杏仁成分或增强（协同）、或减弱（拮抗）麻黄的作用，为臣药；麻黄有效成分中，甲基麻黄碱对绝大多数效应指标的影响最强，应是麻黄的主要有效成分。.本研究初步探明了麻黄-杏仁药对疗喘的物质基础、EGFR作用机理及“配伍-成分-效应”之间的变化，深化了对该药对的现代阐释。