网络药理学预测丹红有效组分配伍抗缺血性脑卒中的作用机制及相关实验研究  

Network pharmacology predicts the mechanism and related experimental research on the effective components of Salvia miltiorrhiza and Carthamus tinctorius against cerebral ischemic stroke

作　　者：朱慧渊 苗琦 王江 罗斌 万海同[2] 王文瑄 董炳耀 肖生斌 党珊[5] ZHU Huiyuan;MIAO Qi;WANG Jiang;LUO Bin;WAN Haitong;WANG Wenxuan;DONG Bingyao;XIAO Shengbin;DANG Shan(School of Basic Medicine, Shaanxi University of Chinese Medicine, Xianyang 712046;School of Biological Engineering, Zhejiang Chinese Medical University, Hangzhou 310053;Department of Pain, The Second Affiliated Hospital of Xi’an Medical College, Xi’an 710068;Department of Statistics, Xi’an Jiaotong University Health Science Center, Xi’an 710061;Department of Geratology, Shaanxi Provincial People’s Hospital, Xi’an 710068, China)

机构地区：[1]陕西中医药大学基础医学院,陕西咸阳712046 [2]浙江中医药大学生物工程学院,浙江杭州310053 [3]西安医学院第二附属医院疼痛科,陕西西安710068 [4]西安交通大学医学部统计学教研室,陕西西安710061 [5]陕西省人民医院老年病科,陕西西安710068

出　　处：《西安交通大学学报（医学版）》2021年第3期474-483,共10页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81503491、81630105、81874366);陕西中医药大学第二附属医院创新团队研究项目(No.2020XKTD-C01)。

摘　　要：目的基于网络药理学筛选丹参-红花药对配伍抗缺血性脑卒中的成分靶点、作用靶点,并对大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)大鼠模型进行相关作用靶点的实验验证。方法①通过系统药理学平台(Traditional Chinese Medicine Systems Pharmacology,TCMSP)、基因名片(GeneCards)数据库筛选丹参-红花的有效成分、成分靶点及缺血性脑卒中的相关靶点;运用STRING数据库构建蛋白互作网络图(protein protein interaction network,PPI),并利用生物信息分析软件(Cytoscape3.8.0)构建成分靶点互作网络;利用David在线分析工具对目标基因进行基因本体论(gene ontology,GO)富集分析和基因的京都百科全书和基因组(kyoto encyclopedia of genes and genomes,KEGG)通路分析。②采用改良MCAO法制备缺血性脑卒中大鼠模型,采用免疫组化法、实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠脑组织中NLR家族Pyrin域蛋白3(NLRP3)炎症小体和核转录因子-B(NF-kB)p65(NFkBp65)蛋白的阳性表达,探讨丹红配伍抗脑缺血损伤炎症反应的作用机制。结果①丹参-红花药对中筛选出87个有效成分,253个靶点,缺血性脑卒中疾病靶点1448个,药物与疾病相关靶点161个,通过GO分析得出730个生物学过程(biological process,BP),81个细胞组分(cellular component,CC),128个分子功能(molecular function,MF),通过KEGG分析得出127条信号通路。②免疫组化法和Real-time PCR测定结果显示,与假手术组比较,模型组大鼠脑组织中NLRP3炎症小体和NFkBp65蛋白表达明显增加(P<0.01),与模型组比较,丹红配伍组和尼莫地平组NLRP3炎症小体和NFkBp65蛋白表达明显降低(P<0.01)。结论丹红有效组分配伍能够通过NF-κB信号通路下调NLRP3炎症小体的释放,阻断炎症损伤环节,最终达到抗炎损伤的作用。Objective To explore the effect mechanism of Salvia miltiorrhiza and safflower on combined anti-ischemic stroke and verify relevant action targets in middle cerebral artery occlusion(MCAO)rat model based on network pharmacology.Methods①Traditional Chinese Medicine Systems Pharmacology(TCMSP)and GeneCardsdatabases were used to screen the active components,component targets and ischemic stroke targets of Salvia miltiorrhiza and safflower respectively.The above data were imported into STRING database for protein interaction network analysis,and Cytoscape3.8.0 software was used to construct protein interaction network(PPI)and component target interaction network.Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway annotation analysis of target genes were performed using David online analysis tool.②In this experiment,a rat model of ischemic stroke was prepared by using improved MCAO method,and immunohistochemical method and Real-time quantitative polymerase chain reaction(REAL-TIME PCR)to detect the positive expressions of NLRP3 inflammatory body and NF P65 protein in the brain tissue of rats in each group so as to explore the functional mechanism of anti-inflammation reaction against cerebral ischemia injury.Results①A total of 87 effective components,corresponding to 253 targets,1448 targets for ischemic stroke and 161 targets related to drugs and diseases,were screened from the Salvia milticorrhiza and safflower drug pairs.We obtained 730 biological processes,81 cell components and 128 molecular functions through GO analysis,and 127 signal pathways through KEGG analysis.②Immunohistochemical method and Real-time PCR determination results showed that compared with control group rats,model group rats had significantly increased tissue NLRP3 inflammatory body and NFkBp65 protein expressions(P<0.01).Compared with those in the model group,NLRP3 inflammatory body and NFkBp65 protein expressions significantly decreased in Dan red compatibility groups and nim horizon groups(P<

关 键 词：网络药理学 丹参 红花 缺血性脑卒中 作用机制 

分 类 号：R289[医药卫生—方剂学]