基于整合药理学策略的银黄清肺胶囊作用解析及研究

发布时间：2018-06-27 18:40

本文选题：整合药理学 + 银黄清肺胶囊　；参考：《北京中医药大学》2017年硕士论文

【摘要】：目的:银黄清肺胶囊是一个含有14味中药的经典名方,其广泛运用于治疗慢性支气管炎并且含有大量的化学成分。然而,系统研究银黄清肺胶囊的化学成分是一个巨大的挑战,目前对该复方中含有的化学成分依然不够清晰。与此同时,虽然银黄清肺胶囊治疗慢性支气管炎在临床已取得了良好的效果,但是其药效基础和作用机理仍未明确。为了对银黄清肺胶囊所含复杂化学成分有一个清晰的认识,并探索银黄清肺胶囊治疗慢性支气管炎的作用机制,运用整合药理学策略对银黄清肺胶囊进行初步解析,为中药复方研究的模式提供新的思路。方法:基于超高效液相色谱串联线性离子阱轨道阱质谱技术建立灵敏、高效的分析方法鉴定银黄清肺胶囊中的化学成分;运用Medchem Studio对银黄清肺胶囊所含的成分的二维结构与Drugbank数据库中的药物进行靶标预测,并对不同单味药之间含有的共同靶标数进行计算;基于中医理论对各单味药所含靶标进行功能富集,构建"中药-功效-药理作用-通路"作用网络,探索中药传统功效与现代药理作用之间的关联;收集与慢性支气管炎相关的疾病靶标分子,构建银黄清肺胶囊成分靶标-疾病靶标相互作用网络,并根据"连接度"、"节点介数"、"紧密度'"、"核心度"四个拓扑结构特征值筛选关键节点,并对关键节点进行通路富集分析,构建"中药-靶标-通路"作用网络,找到银黄清肺胶囊发挥疗效可能参与的生物过程,并明确相应关键靶标对应的化合物。最后用分子对接的手段对分析结果模拟验证,通过考察小分子化合物与靶标蛋白的结合能力,间接验证网络药理学预测的可靠性。结果:1.根据多级质谱数据,色谱行为,标准品和以前的文献报道,一共在银黄清肺胶囊中鉴定了 204个成分,包括黄酮类化合物68个,生物碱类化合物25个,木脂素类化合物22个,有机酸及其酯类化合物22个,三萜类化合物17个,甾体皂苷类化合物11个,香豆素类化合物11个,萜类化合物6个,柠檬苦素类化合物5个,其他化合物17个。这些化合物的药材来源也进行了分析。其中,37个成分来源于甘草,29个成分来源于银杏叶,27个成分来源于枳实,24个成分来源于五味子,15个成分来源于枇杷叶,14个成分来源于浙贝母,13个成分来源于新疆一枝蒿,12个成分来源于大青叶,12个成分来源于穿山龙,10个成分来源于蜜麻黄,10个成分来源于苦杏仁,8个成分来源于石菖蒲,6个成分来源于北葶苈子。2.银黄清肺胶囊成分靶标预测共得到26917个"成分-靶标"对,不同单味药之间含有不同数目的相同靶标,提示不同单味药之间可能具有协同或拮抗作用,而各单味中药与银杏叶之间的共同靶标数均大于其总靶标数的50%,在一定程度上说明银杏叶与其他各味中药之间的关联是比较紧密的。3.通过"中药-功效-药理作用-通路"相互作用网络构建,对单味药成分靶标聚集的通路分析,发现中药功效与现代药理活性密切相关。对银黄清肺胶囊成分靶标-疾病靶标相互作用网络分析,经筛选,共得到475个关键靶标。经通路富集分析,发现关键靶标显著富集在炎症-免疫、支气管平滑肌收缩和神经中枢等功能模块,而在相关通路中,多个病理过程均参与到Asthma通路,而聚集到Asthma通路上的IL-3,IL-4,IL-5,IL-10,IL-13,FCER1G,CCL11和EPX这8个分子与慢性支气管炎的发病过程密切相关,被认为是银黄清肺胶囊治疗慢性支气管炎过程中所调控的关键靶标分子。分子对接结果显示,有17对小分子化合物与靶标分子具有强结合能力。结论:运用液质联用技术共鉴别了 204个化合物,为银黄清肺胶囊进一步的质量控制研究,药理活性预测研究做良好的铺垫;运用网络药理学技术进行银黄清肺胶囊靶标预测和网络构建,在计算水平上阐释了银黄清肺胶囊作用于慢性支气管炎的分子机制。本文的研究特色及创新点:1.基于整合药理学策略运用超高效液相-高分辨质谱联用技术对中药大复方银黄清肺胶囊所含化学成分进行了全面解析,共鉴定了 204个化学成分;2.基于整合药理学策略运用网络药理学预测技术对中药大复方银黄清肺胶囊中的关键成分和靶标的相互作用关系进行网络构建,并对关键成分、靶标及通路进行筛选分析,阐释银黄清肺胶囊作用于慢性支气管炎的分子机制,并运用分子对接技术进行模拟验证。
[Abstract]:Objective: Yinhuang Qingfei capsule is a classic prescription containing 14 flavors, which is widely used in the treatment of chronic bronchitis and contains a large number of chemical components. However, the chemical composition of Yinhuang Qingfei capsule is a huge challenge, and the chemical composition in the compound is still not clear. At the same time, although the chemical composition of the compound is still not clear. But Yinhuang Qingfei capsule has achieved good results in the treatment of chronic bronchitis, but the basis and mechanism of its efficacy are still not clear. In order to understand the complex chemical components of Yinhuang Qingfei capsule, the mechanism of Yinhuang Qingfei capsule in the treatment of chronic bronchitis is explored, and the integrated pharmacology strategy is used. The preliminary analysis of Yinhuang Qingfei capsule provides a new idea for the model of Chinese medicine compound research. Method: a sensitive and efficient analysis method based on super high performance liquid chromatography tandem linear well track trap mass spectrometry is established to identify the chemical components in Yinhuang Qingfei capsule, and Medchem Studio is used for the formation of Yinhuang Qingfei capsule. The two dimensional structure and the drugs in the Drugbank database are predicted, and the number of common targets among the different single flavors is calculated. Based on the theory of traditional Chinese medicine, the targets are enriched and the "Chinese medicine efficacy pharmacological action pathway" network is constructed to explore the traditional and modern pharmacological effects of traditional Chinese medicine. To collect the target molecules of the disease related to chronic bronchitis, construct the interaction network of Yinhuang Qingfei capsule component target disease target, and select the key nodes according to the "connectivity", "node number", "tightness", "core degree" four topological structure characteristic values, and analyze the key nodes through the path enrichment analysis, and construct "Chinese Medicine -" The target channel "Action Network" is used to find the biological processes that may be involved in the effect of Yinhuang Qingfei capsule, and to clarify the corresponding compounds corresponding to the corresponding key targets. Finally, the analysis results are simulated and verified by the means of molecular docking, and the reliability of the network pharmacology prediction is indirectly verified by investigating the binding ability of the small molecular compound to the target protein. Results: 1. according to multilevel mass spectrometry data, chromatographic behavior, standard products and previous literature reports, a total of 204 components were identified in Yinhuang Qingfei capsule, including 68 flavonoids, 25 alkaloids, 22 lignans, 22 organic acids and their esters, 17 three terpenoids, steroid saponins. There are 11 compounds, 11 coumarins, 6 terpenoids, 5 citrin compounds and 17 other compounds. The sources of these compounds are also analyzed. Among them, 37 components are derived from licorice, 29 are derived from Ginkgo biloba leaves, 27 are derived from Fructus aurantii, 24 components are derived from Schisandra chinensis and 15 sources. In loquat leaf, 14 ingredients come from Fritillaria thunbergii, 13 components come from Artemisia Artemisia in Xinjiang, 12 of them originate from the leaves of dacea, 12 from the Dioscorea, 10 from Ephedra, 10 from bitter almond and 8 from Acorus calamus. 6 components are derived from the target prediction of the.2. Yinhuang Qingfei capsule. A total of 26917 "composition target" pairs were obtained, with different single flavors containing the same target number, suggesting that different single flavors may have synergistic or antagonistic effects. The common target number of each single flavored Chinese medicine and Ginkgo biloba leaves is more than 50% of the total target number. The correlation is a relatively close.3. through the "traditional Chinese medicine efficacy pharmacological action pathway" interaction network construction, the analysis of the pathway of the target aggregation of single flavors, found that the efficacy of traditional Chinese medicine is closely related to the modern pharmacological activity. The analysis of the interaction network of the target target disease target of Yinhuang Qingfei capsule has been screened, and 475 key targets are obtained. By pathway enrichment analysis, it was found that the key targets were significantly enriched in the function modules of inflammatory immunity, bronchial smooth muscle contraction and nerve center, while in the related pathways, multiple pathological processes were involved in the Asthma pathway, and the 8 molecules of IL-3, IL-4, IL-5, IL-10, IL-13, FCER1G, CCL11, and EPX, on the Asthma pathway, and chronic bronchi The process of inflammation is closely related and is considered to be the key target molecule controlled by Yinhuang Qingfei capsule in the treatment of chronic bronchitis. Molecular docking results show that there are 17 pairs of small molecules with strong binding ability to the target molecule. Conclusion: 204 compounds were identified by the technique of liquid chromatography-using yellowish yellow Qingfei capsule. One step of quality control research, pharmacological activity prediction research to make good paving, using network pharmacology to predict the target and network construction of Yinhuang Qingfei capsule, explain the molecular mechanism of Yinhuang Qingfei capsule acting on chronic bronchitis. The characteristics and innovation of this paper: 1. based on integrated pharmacology policy The chemical components of the big compound Yinhuang Qingfei capsule were comprehensively analyzed by super high performance liquid phase high resolution mass spectrometry, and 204 chemical components were identified. 2. based on the integrated pharmacology strategy, the interaction between the key components and target in the big compound silver Huang Qingfei capsule was made by using the network pharmacology prediction technology. The key components, targets and pathways were screened and analyzed by using the relationship. The molecular mechanism of Yinhuang Qingfei Capsule on chronic bronchitis was explained, and the molecular docking technique was used to simulate and verify the molecular mechanism.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R286.0;R285

【参考文献】