基于网络药理学的肉苁蓉苯乙醇苷抗氧化活性分子机制研究

目的 运用网络药理学方法探讨肉苁蓉苯乙醇苷类成分抗氧化的分子作用机制，并采用体外实验对肉苁蓉中主要苯乙醇苷的抗氧化活性及构效关系进行初步研究。方法 通过TCMSP数据库和文献挖掘，获取分布于中国的5种肉苁蓉属植物中的苯乙醇苷类化合物，利用PharmMapper服务器、CoolGeN和GeneCards数据库预测和筛选苯乙醇苷抗氧化作用靶点，采用Cytoscape 3.6.1构建成分-靶点网络。使用String数据库进行蛋白-蛋白相互作用分析，借助DAVID数据库对靶点进行GO分析和KEGG通路分析，采用Cytoscape 3.6.1构建药材-成分-靶点-通路-疾病网络图。采用DPPH自由基清除法和普鲁士蓝法，测定肉苁蓉苯乙醇苷、苷元和结构类似物的抗氧化能力和总还原能力；采用分子对接评价上述成分与重要靶点的结合能力，为药物的靶点预测和验证提供基础。结果 筛选得到苯乙醇苷类化合物共71个，潜在作用靶点114个，进一步筛选抗氧化相关靶点，发现谷胱甘肽S转移酶P1(glutathione S-transferase P1，GSTP1)、表皮生长因子受体(epidermal growth factor receptor，EGFR)等44个靶点与抗氧化相关；KEGG通路分析显示，上述靶点涉及催乳素信号通路、FoxO信号通路、Fc epsilon RI信号通路等51条通路。4种苯乙醇苷、苷元和结构类似物均能够清除DPPH自由基，具有较强的还原力，并且与靶点结合能力较强。结论 苯乙醇苷可以通过调节GSTP1、EGFR、MAPK1等靶点发挥抗氧化作用；苯乙醇苷的抗氧化作用与结构中的咖啡酰和3,4-二羟基苯乙醇基团有关。

Molecular Mechanism of Antioxidant Activity of Phenylethanoid Glycosides from Cistanches Herba Based on Network Pharmacology

OBJECTIVE To explore the antioxidant molecular mechanism of phenylethanoid glycosides from Cistanches Herba by employing the network pharmacological method, as well as to explore the antioxidant activity and structure-activity relationship of the main phenylethanoid glycosides by in vitro experiments. METHODS Phenylethanoid glycosides from 5 Cistanches Herba distributed in China were obtained by TCMSP database and literature search. Pharmmapper web server, CoolGeN and GeneCards database were used to predict and screen the antioxidant targets of the above components and the ingredient-target network was subsequently constructed by Cytoscape 3.6.1. The protein-protein interaction network was constructed by the String database. The GO and KEGG pathway analysis of the targets were analyzed by the DAVID database. The ingredient-target-pathway-disease network was constructed by Cytoscape 3.6.1. DPPH radical scavenging method and Prussian blue spectrophotometry were used to evaluate the antioxidant capacity and total reducing capacity of phenylethanoid glycosides, aglycons and structural analogs, the molecular docking was adopted to evaluate the binding ability of the above compounds and important targets, providing a basis for target prediction and validation of the ingredients. RESULTS It was found that 71 phenylethanoid glycosides active components and 114 targets were involved. A total of 44 antioxidant-related targets were further screened, including glutathione S-transferase P1(GSTP1), epidermal growth factor receptor(EGFR). KEGG pathway analysis showed that the above antioxidant-related targets were enriched in 51 pathways, such as the prolactin signaling pathway, FoxO signaling pathway, and Fc epsilon RI signaling pathway. Four phenylethanoid glycosides, aglycons and structural analogs had the ability to scavenge DPPH free radicals and strong reducing power, also had strong binding ability with the targets. CONCLUSION Phenylethanoid glycosides can exert antioxidant effects by modulating GSTP1, EGFR, MAPK1 and other targets. The antioxidant activities of phenylethanoid glycosides are related to caffeoyl and 3,4-dihydroxyphenylethanol groups in the structure.