基于网络药理学和动物实验探讨桃红四物汤抗缺血性脑卒中的分子机制  

Molecular mechanism of Taohong Siwu decoction against ischemic stroke based on network pharmacology and animal experiment

作　　者：王艺霖 孟祥国 叶宇航 胡霞敏 WANG Yi-lin;MENG Xiang-guo;YE Yu-hang;HU Xia-min(Graduate School of Shanghai University of Traditional Chinese Medicine,Shanghai 201203;Shanghai University of Medicine&Health Sciences,Shanghai 201318;Shanghai University of Science and Technology,Shanghai 200093)

机构地区：[1]上海中医药大学研究生院,上海201203 [2]上海健康医学院,上海201318 [3]上海理工大学,上海200093

出　　处：《中南药学》2024年第2期392-398,共7页Central South Pharmacy

摘　　要：目的通过网络药理学、分子对接技术和动物实验探讨桃红四物汤对缺血性脑卒中的调控机制。方法运用数据库整理分析桃红四物汤与疾病的交集靶点,进行网络拓扑分析及蛋白质相互作用分析并获取核心成分和核心靶点;对交集靶点进行GO和KEGG富集分析;利用分子对接技术获得通路蛋白与核心成分结合能;采用线拴法建立小鼠脑缺血再灌注损伤(MCAO/R)模型,利用神经行为学评分、TTC染色技术考察桃红四物汤防治缺血性脑卒中的药效作用,利用Western blot技术考察桃红四物汤对MCAO/R小鼠脑中PI3K、AKT蛋白和炎症因子IL-1β、IL-18表达的影响。结果桃红四物汤抗缺血性脑卒中的核心成分可能为槲皮素、木犀草素、β-谷甾醇、山柰酚、豆甾醇、黄芩素、杨梅酮、β-胡萝卜素及常春藤苷,核心靶点可能为MAPK14、RELA、TNF、TR53、MAPK1、MYC、FOS、HSP9AA1、ESR1、AKT1、HF1A、CTNNB1。交集靶点主要富集在脂质和动脉粥样硬化、PI3K/AKT等信号通路,涉及氧化应激、对脂多糖的反应等生物进程。分子对接预测核心成分与通路蛋白具有较好的结合能力。动物实验表明桃红四物汤可降低MCAO/R小鼠神经功能评分,减小脑梗死体积,降低脑内IL-1β、IL-18的表达,提高PI3K、AKT的表达。结论桃红四物汤可能通过激活PI3K/AKT信号通路,调节神经炎症,防治缺血性脑卒中。Objective To determine the mechanism of Taohong Siwu decoction(THSWD)in ischemic stroke by network pharmacology,molecular docking and in vivo experiment.Methods The common targets of THSWD and disease were analyzed via database collection.Network topology analysis and protein interaction analysis were used to obtain the key components and key targets.The common targets were analyzed by GO and KEGG enrichment.The binding energy of related pathway proteins and key components was obtained by molecular docking.The mouse focal ischemia-reperfusion injury model was established by ligation of middle cerebral artery occlusion reperfusion(MCAO/R).The effect of THSWD on stroke was determined by neurobehavioral tests and TTC staining.Western blot was used to verify the effect of THSWD on the expression of IL-1β,IL-18,PI3K,and AKT in the middle MCAO/R mouse brain.Results The key components of THSWD on ischemic stroke might include quercetin,luteolin,β-sitosterol,kaempferol,stigmasterol,baicalin,myricetin,β-carotene and ivioside.THSWD and the key potential targets of disease included MAPK14,RELA,TNF,TR53,MAPK1,MYC,FOS,HSP9AA1,ESR1,AKT1,HF1A and CTNNB1.The common targets were mainly concentrated in the lipid and atherosclerosis signaling pathways,and PI3K/AKT signaling pathways,which involved in such biological processes as oxidative stress and response to lipopolysaccharide.The molecular docking showed that key components had strong binding affinity to pathway protein.The in vivo experiment assays showed that THSWD decreased the neural function score,reduced the cerebral infarction area,decreased the expression levels of IL-1βand IL-18,and increased the expression levels of PI3K and AKT in the MCAO/R mouse brain.Conclusion THSWT may prevent stroke via the activation of PI3K/AKT signaling pathway to regulate neuritis.

关 键 词：网络药理学 分子对接 桃红四物汤 脑卒中 分子机制 

分 类 号：R285.5[医药卫生—中药学]