基于UPLC-Q-Orbitrap HRMS和网络药理学分析五味子治疗非酒精性脂肪肝的物质基础及作用机制

任媛媛; 王超; 谢勇; 郭光鹏; 黄勤挽; 王瑾

doi:10.13386/j.issn1002-0306.2021090347

基于UPLC-Q-Orbitrap HRMS和网络药理学分析五味子治疗非酒精性脂肪肝的物质基础及作用机制

UPLC-Q-Orbitrap HRMS Combined with Network Pharmacology to Analyze the Material Basis and Mechanism of Schisandrae chinensis in the Treatment of Non-alcoholic Fatty Liver

摘要

摘要: 目的：基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱法（UPLC-Q-Orbitrap HRMS）、网络药理学方法及分子对接技术探索五味子治疗非酒精脂肪肝的物质基础与作用机制。方法：利用UPLC-Q-Orbitrap HRMS技术鉴定五味子中的化学成分。利用中药系统药理学数据库和分析平台（TCMSP）和SwissTarget Prediction在线平台筛选并预测五味子化学成分的潜在作用靶点；利用GeneCards，CTD，OMIM，Dis-GeNET，GEO数据库收集非酒精脂肪肝疾病的潜在作用靶点。利用String数据库和Cytoscape 3.7.1软件构建蛋白质-蛋白质相互作用（PPI）网络模型；使用R对潜在靶点进行基因本体论（gene ontology，GO）和基因组百科全书通路（kyoto encyclopedia of genes and genomes，KEGG）富集分析，以预测其可能的信号通路，并通过Cytosca-pe3.7.1软件建立“药物-关键活性成分-靶点途径”网络。最后，进行分子对接，以初步验证五味子治疗非酒精性脂肪肝的作用机制。结果：共分析鉴定了五味子中的50种化学成分，并以此为基础筛选出246个五味子治疗非酒精脂肪肝的潜在靶点。通过网络拓扑进一步分析，筛选出23个核心成分和30个潜在核心靶点。GO和KEGG富集分析发现五味子通过作用于癌症中的蛋白聚糖、内分泌抵抗、Rap1信号通路、VEGF信号通路、糖尿病并发症中的AGE-RAGE信号通路和雌激素信号通路等信号通路发挥治疗作用。分子对接结果显示，筛选出的排名前5的活性成分槲皮素、山奈酚、五味子甲素、α-亚麻酸和五味子醇乙与排名前4的核心靶点AKT1、HSP90AA1、SRC、MAPK1均具有较好的结合活性，结合自由能均小于-5 kcal/mol，分子对接构象稳定。结论：五味子可能通过作用于脂质代谢、氧化应激、血管新生和炎症相关途径和靶点等改善非酒精脂肪肝的症状，从而发挥作用。

Abstract: Objective: UPLC-Q-Orbitrap HRMS, network pharmacology and molecular docking technology were used to explore the material basis and mechanism of Schisandrae chinensis (SC) in treating non-alcoholic fatty liver disease(NAFLD). Method: UPLC-Q-Orbitrap HRMS was used to identify the chemical components in SC. The Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP) and Swiss Target Prediction online platform were used to screen and predict the potential targets of the chemical composition of SC; GeneCards, CTD, OMIM, Dis-GeNET, GEO database were used to collect non-alcoholic fatty liver disease potential targets. String database and Cytoscape 3.7.1 software were used to construct a protein-protein interaction (PPI) network model; R-based bioconductor data package for gene ontology (GO) and KEGG pathway were used to analyse potential targets; Cytoscape 3.7.1 software were used to establish a "drug-key active ingredient-target-pathway" network. Finally, molecular docking was carried out to preliminarily verify the mechanism of SC in the treatment of NAFLD. Results: 50 components of SC were analyzed and identified, and 246 potential targets of SC in treating NAFLD were screened out. Through further analysis of network topology, 23 core components and 30 potential core targets were screened out, and enrichment analysis was carried out based on them. SC played a therapeutic role through the proteoglycans in cancer, endocrine resistance, Rap1 signaling pathway, VEGF signaling pathway, AGE-RAGE signaling pathway in diabetic complications and Estrogen signaling pathway. Molecular bonding results showed that the top 5 active ingredients, quercetin, kaempferol, schisandrin a, α-linolenic acid and schisandrol B, had good binding activities with the top 4 core targets, AKT1, HSP90AA1, SRC and MAPK1, and their binding free energies were all less than −5 kcal/mol. The docking conformation of the molecule was stable. Conclusion: SC may improve the symptoms of NAFLD by acting on lipid metabolism, oxidative stress, angiogenesis and inflammation related pathways and targets.