基于网络药理学和实验验证探讨熄风清火活血化痰方治疗缺血性脑卒中的作用机制  

Mechanism of Xifeng Qinghuo Huoxue Huatan Decoction in Treating Ischemic Stroke Based on Network Pharmacology

作　　者：蒲馨颖 英振昊[2] 郭大千 门伟业 朱文浩[3] PU Xin-ying;YING Zhen-hao;GUO Da-qian;MEN Wei-ye;ZHU Wen-hao(Shandong University of Traditional Chinese Medicine,Jinan Shandong 250355;School of Rehabilitation Medicine,Shandong University of Traditional Chinese Medicine,Jinan Shandong 250355;Department of Encephalopathy,Zibo Hospital of Traditional Chinese Medicine,Zibo Shandong 255300)

机构地区：[1]山东中医药大学,山东济南250355 [2]山东中医药大学康复医学院,山东济南250355 [3]山东淄博市中医医院脑病科,山东淄博255300

出　　处：《世界中西医结合杂志》2023年第6期1178-1185,共8页World Journal of Integrated Traditional and Western Medicine

基　　金：山东省中医药科技项目(2020M114);淄博市重点研发计划项目(2019ZC010124);第五批全国中医临床优秀人才研修项目(国中医药人教函[2022]1号)。

摘　　要：目的基于网络药理学方法探讨熄风清火活血化痰方治疗缺血性脑卒中(Ischemic stroke,IS)潜在的作用机制,并对其潜在的功能靶点进行实验验证。方法采用TCMSP平台、ETCM平台、TCMID平台、BATMAN-TCM、文献筛选熄风清火活血化痰方中各中药的活性成分,利用TCMSP靶点预测模型、STITCH、SwissTarget、SEA数据库预测活性成分的潜在靶点;利用Genecards、OMIM、Disgenet数据库检索缺血性卒中的相关靶点并筛选得到疾病靶点,取疾病靶点与药物靶点的交集,利用STRING 11.0对药物与疾病的交集靶点进行蛋白质相互作用网络进行构建,通过Cytoscape 3.6.0软件进行拓扑分析获得熄风清火活血化痰方治疗缺血性卒中的关键靶点;利用Metascape对关键靶点进行GO和KEGG富集分析。通过Western blot检测对网络药理学的富集分析结果进行验证。结果共得到有效成分203个,药物靶点3971个,筛选缺血性脑卒中靶点1798个,最终获得关键靶点60个,KEGG通路富集分析筛选得到前20条信号通路。熄风清火活血化痰方治疗缺血性卒中主要成分为槲皮素、山奈酚、β-谷甾醇、木樨草素、豆甾醇等,关键靶点AKT1、TP53、IL-6、MAPK1、MAPK8等。关键的信号通路可能包括AGE-RAGE信号通路、流体剪切力与动脉粥样硬化信号通路、NF-kB信号通路、PI3k-Akt信号通路、VEGF信号通路等。Western blot检测结果表明,与空白组比较,模型组p-AKT1、p-P53、p-JNK1、p-ERK2表达升高,差异有统计学意义(P<0.05),与模型组比较,中药组p-AKT1、p-P53、p-JNK1、p-ERK2、IL-6表达降低,AKT1表达升高,差异有统计学意义(P<0.05)。结论揭示了熄风清火活血化痰方多成分、多靶点、多途径治疗缺血性卒中的机制,初步证实了该方对靶点蛋白AKT1、TP53、IL-6、MAPK1、MAPK8表达的影响,为熄风清火活血化痰方的临床开发利用提供了依据。Objective To explore the potential mechanism of Xifeng Qinghuo Huoxue Huatan Decoction in treating ischemic stroke(IS)based on network pharmacology and experimentally verify the potential functional targets.Methods The active components of the Chinese medicines in Xifeng Qinghuo Huoxue Huatan Decoction were obtained from TCMSP,ETCM,TCMID,BATMAN-TCM,and the available literature.The potential targets of the active components were predicted via the target predictive model of TCMSP,STITCH,SwissTarget,and SEA.Ischemic stroke-related targets were retrieved from Genecards,OMIM,and Disgenet,and the common targets shared by the disease and the decoction were identified.STRING 11.0 was employed to establish the protein-protein interaction network of the common targets,and Cytoscape 3.6.0 to predict the key targets of the decoction for treating this disease.Metascape was used for GO functional annotation and KEGG pathway enrichment on the key targets.The prediction results of network pharmacology were verified by Western blotting.Results There were 203 active components in Xifeng Qinghuo Huoxue Huatan Decoction,which corresponded to 3971 targets,and 1798 targets for ischemic stroke were obtained.Finally,60 key targets were obtained and the top 20 signaling pathways were screened out by the enrichment analysis.The main components of Xifeng Qinghuo Huoxue Huatan Decoction in treating ischemic stroke were quercetin,kaempferol,beta-sitosterol,luteolin,stigmasterol,etc.The key targets were AKT1,TP53,IL-6,MAPK1,MAPK8,etc.AGE-RAGE,fluid shear stress and atherosclerosis,PI3k-Akt,NF-kB,and VEGF signaling pathways may be the key ones.Western blot showed that compared with the blank group,the model group showed up-regulated expression of p-AKT1,p-P53,p-JNK1,and p-ERK2(P<0.05).Compared with the model group,the decoction down-regulated the expression of p-AKT1,p-P53,p-JNK1,p-ERK2,and IL-6 and up-regulated the expression of AKT1(P<0.05).Conclusion This study preliminarily revealed the multi-component,multi-target,and multi

关 键 词：网络药理学 熄风清火活血化痰方 缺血性脑卒中 

分 类 号：R285.6[医药卫生—中药学]