基于网络药理学和分子对接技术探讨黄芪-紫苏子配伍治疗慢性阻塞性肺疾病的作用机制＊

目的 采用网络药理学和分子对接技术研究黄芪-紫苏子配伍治疗慢性阻塞性肺疾病（COPD）的主要化学成分、可能的作用靶点以及相关信号通路,探讨其多成分-多靶点-多通路的潜在作用机制。方法 通过TCMSP数据库检索黄芪-紫苏子的化学成分;运用PubChem数据库和Swiss Target Prediction数据库筛选其潜在靶点;借助GeneCards数据库检索COPD相关靶点,与黄芪-紫苏子化合物靶点进行交集筛选出共同靶点作为研究靶点;使用Cytoscape3.2.1软件构建“中药-化学成分-靶点-疾病”网络;将上述共同靶点导入String数据库获取数据后在Cytoscape3.2.1软件中构建蛋白质-蛋白质相互作用网络（protein-protein interaction,PPI）,然后进行核心靶点筛选;利用Metascape数据库进行GO和KEGG通路富集分析;使用RCSB PDB数据库、Pymol和Autodock Vina软件进行分子对接;采用A549细胞进行体外活性实验评价槲皮素和异鼠李素的抗炎作用,验证网络药理学及分子对接结果的科学准确性。结果 黄芪-紫苏子的“中药-化学成分-靶点-疾病”网络包含34个化学成分和74个治疗COPD的潜在靶点,其核心化学成分主要为黄酮类化合物;另PPI网络中核心靶点主要涉及ALB、TP53、AKT1等;GO功能富集得到GOBP条目1841条,GOCC条目109条,GOMF条目105条;KEGG通路富集筛选得到130条信号通路,其中靶点在AGE-RAGE信号通路、PI3K-AKT 信号通路和HIF-1信号通路等富集较为集中。分子对接结果显示,黄芪和紫苏子的核心化合物中的黄芪异黄烷苷、木犀草素、槲皮素以及异鼠李素等黄酮类成分对ALB具有较高亲和力。体外活性验证中槲皮素和异鼠李素具有显著的抗炎活性。结论 黄芪-紫苏子配伍可通过多成分、多靶点和多通路调控炎症因子释放,达到治疗COPD的效果,为其分子机制研究奠定一定的基础。

Study on the Mechanism of Radix Astragali seu Hedysari-Fructus Perillae Combination in the Treatment of Chronic Obstructive Pulmonary Disease Based on Network Pharmacology and Molecular Docking

Objective To explore the potential mechanism of the multicomponent-multitarget-multi pathway action of the Radix Astragali seu Hedysari-Fructus Perillae combination in the treatment of chronic obstructive pulmonary disease (COPD) by using network pharmacology and molecular docking and to investigate the main chemical components, possible targets of action and related signaling pathways.Methods The chemical components of Radix Astragali seu Hedysari-Fructus Perillae were searched through TCMSP database; the potential targets were screened through PubChem database and Swiss Target Prediction database; COPD-related targets were searched through GeneCards database, and the common targets were screened through intersection with the targets of Astragali seu Hedysari-Fructus Perillae compounds as research targets; the common targets were constructed using Cytoscape 3.2.1 software. Cytoscape3.2.1 software was used to construct a "medicinal-chemical component-target-disease" network; the above common targets were imported into the String database to obtain data and then a protein-protein interaction (PPI) network was constructed in Cytoscape3.2.1 software. The above common targets were imported into String database to obtain data and then constructed protein-protein interaction (PPI) network in Cytoscape 3.2.1 software, followed by core target screening; GO and KEGG pathway enrichment analysis using Metascape database; molecular docking using RCSB PDB database, Pymol and Autodock Vina software; in vitro activity assay used A549 cells to evaluate the anti-inflammatory effects of quercetin and isorhamnetin. The anti-inflammatory effects of quercetin and isorhamnetin were evaluated in vitro using A549 cells to verify the scientific accuracy of the network pharmacology and molecular docking results.Results The "medicinal-chemical component-target-disease" network of Astragali seu Hedysari-Fructus Perillae contained 34 chemical components and 74 potential targets for the treatment of COPD, with the core chemical components mainly being flavonoids; the core targets in the PPI network mainly involved ALB, TP53, AKT1, etc.; GO function enrichment yielded GOBP entries 1841, GOCC entries 109, GOMF entries 105; KEGG pathway enrichment screening obtained 130 signaling pathways, of which targets in AGE-RAGE signaling pathway, PI3K-AKT signaling pathway and HIF-1 signaling pathway were more concentrated. The molecular docking results showed that the flavonoid components of Astragali seu Hedysari-Fructus Perillae, such as isomucronulatol 7-O-glucoside, luteolin, quercetin and isorhamnetin, had high affinity for ALB. In vitro activity was verified in which quercetin and isorhamnetin showed significant anti-inflammatory activity.Conclusion Astragali seu Hedysari-Fructus Perillae combination can regulate the release of inflammatory factors through multi-component, multi-target and multi-pathway to achieve the effect of treating COPD, laying a certain foundation for the study of its molecular mechanism.