基于网络药理学、分子对接及实验验证探讨当归挥发油改善血管性痴呆的作用机制  

Exploration of the Mechanism of Volatile Oil of Angelicae Sinensis Radix in Improving Vascular Dementia Based on Network Pharmacology,Molecular Docking and Experimental Validation

作　　者：范月盈 邓翠莉 张力升 罗晶 张韧[1] 刘娜[1] 刁远明[1] FAN Yueying;DENG Cuili;ZHANG Lisheng;LUO Jing;ZHANG Ren;LIU Na;DIAO Yuanming(Guangzhou University of Chiness Medicine,Guangzhou 510006 Guangdong,China;Shenzhen Hospital of Integrated Traditional and Western Medicine,Shenzhen 518000 Guangdong,,China)

机构地区：[1]广州中医药大学,广东广州510006 [2]深圳市中西医结合医院,广东深圳518000

出　　处：《中药新药与临床药理》2023年第8期1117-1128,共12页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省自然科学基金面上项目(2022A1515011575)。

摘　　要：目的基于网络药理学、分子对接及实验验证探讨当归挥发油(Volatile oil of Angelica Sinensis Radix,VOASR)改善血管性痴呆(Vascular dementia,VaD)的作用机制。方法(1)采用气相色谱-质谱联用(GC-MS)法对VOASR的主要化学成分进行分析鉴定,确定百分含量排前5位的化合物。通过Swiss Target Prediction数据库筛选出VOASR 5个主要化学成分的作用靶点,通过OMIM、GeneCards、Drugbank数据库收集VaD疾病相关靶点。对VOASR主要化学成分的作用靶点与VaD疾病相关靶点取交集,得到二者共同靶点即为VOASR干预VaD的潜在作用靶点。运用STRING数据库对交集靶点构建蛋白相互作用(PPI)网络,并分析判断关键靶点。使用R语言的Bioconductor软件包对共同靶点进行GO功能富集分析及KEGG通路富集分析,并使用Cytoscape 3.7.2软件构建VOASR干预VaD的“药物-成分-疾病-靶点-通路”网络。采用AutoDock Vina软件进行分子对接,分析5个主要化学成分与关键靶点蛋白的结合能力,进一步筛选出VOASR治疗VaD的核心靶点。(2)体内实验:将C57BL/6小鼠随机分为假手术组、模型组、VOASR组(105 mg·kg^(-1)),每组6只。采用单侧颈总动脉结扎术(UCCAO)构建VaD小鼠模型;术后灌胃给药(10 mL·kg^(-1)),每日1次,持续3周。采用Morris水迷宫实验评估VaD小鼠的认知功能;采用免疫组织化学法及Western Blot法检测小鼠海马组织中多巴胺受体D2(DRD2)、单胺氧化酶A(MAOA)蛋白表达情况。结果(1)VOASR中相对百分含量排前5位的核心化学成分分别为(E)-Ligustilide(E-藁本内酯)、Z-Butylidenephthalide(Z-正丁烯基苯酞)、Senkyunolide H(洋川芎内酯H)、Spathulenol(桉油烯醇)、3-Butylphthalide[3-丁基-1(3H)-异苯并呋喃酮],占总含量的97.32%。共筛选得到VOASR中5个主要化学成分的作用靶点92个,VaD疾病相关靶点1181个,得到共同靶点(交集靶点)31个。分析得到VOASR治疗VaD的关键靶点包括VEGFA、PTGS2、ICAM1、VCAM1、F2、FGF2Objective To investigate the mechanism of volatile oil of Angelicae Sinensis Radix(VOASR)in improving vascular dementia(VaD)based on network pharmacology,molecular docking and experimental validation.Methods(1)Gas chromatography-mass spectrometry(GC-MS)was used to analyze and identify the main chemical components of VOASR and determine the top 5 compounds in terms of percentage content.The targets of the 5 major chemical components of VOASR were screened by Swiss Target Prediction database,and the targets related to VaD disease were collected by OMIM,GeneCards and Drugbank databases.The common targets of the main chemical components of VOASR and the disease-related targets of VaD were intersected,and the common targets were identified as the potential targets of VOASR for VaD intervention.A protein-protein interaction(PPI)network was constructed for the intersecting targets using the STRING database,and the key targets were analyzed.GO functional enrichment analysis and KEGG pathway enrichment analysis of common targets were performed using the Bioconductor software package in R language.Cytoscape 3.7.2 was used to construct a"drugs-components-diseases-targets-pathways"network for VOASR interventions in VaD.Molecular docking was performed using AutoDock Vina software to analyze the binding ability of the 5 major chemical components to key target proteins,and to further screen the core targets of VOASR for VaD treatment.(2)In vivo experiments:C57BL/6 mice were randomly divided into sham-operation group,model group and VOASR group(105 mg·kg^(-1)),with 6 mice in each group.The VaD mouse model was constructed by unilateral common carotid artery occlusion(UCCAO);the drug was administered by gavage(10 mL·kg^(-1))once daily for3 consecutive weeks after surgery.The cognitive function of VaD mice was assessed by Morris water maze test;the protein expressions of dopamine receptor D2(DRD2)and monoamine oxidase A(MAOA)in the hippocampal tissue of mice were measured by immunohistochemistry and Western Blot.Results(1)The top

关 键 词：当归挥发油 血管性痴呆 网络药理学 分子对接 E-藁本内酯 洋川芎内酯H 多巴胺受体D2 单胺氧化酶A 小鼠 

分 类 号：R285.5[医药卫生—中药学] R857.3[医药卫生—中医学]