摘要
目的:运用网络药理学方法探讨活络效灵丹治疗缺血性中风的作用机制。方法:利用中医药证候关联数据库(SymMap)、中药综合资源数据库(TCM-ID)、草药成分和靶标数据库(HIT)以及SwissTargetPrediction数据库检索活络效灵丹的活性成分、作用靶点;通过人类基因综合数据库(GeneCards)、比较毒理基因组学数据库(CTD)、基因组学数据库(DisGeNET)、基因表达综合数据库(GEO)等筛查缺血性中风的疾病靶点与差异表达基因;利用UpSetR对活络效灵丹作用靶点、差异表达基因和缺血性中风疾病靶点取交集,筛选出活络效灵丹治疗缺血性中风相关靶点;通过Metascape数据库对相关靶点进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集分析。利用Cytoscape 3.9.10软件构建蛋白质互作网络和药物-活性成分-靶基因网络,筛选出活络效灵丹治疗缺血性中风的核心成分和核心靶点。通过单样本基因集富集分析对GSE16561数据集中正常和缺血性中风样本间的免疫细胞浸润差异进行分析,并分析核心靶点与免疫细胞浸润水平的相关性。采用分子对接验证核心靶点与活性成分的亲和性。结果:共获得活络效灵丹活性成分226个,靶点1541个,缺血性中风靶点712个,差异表达基因3136个。取交集后,共获得活络效灵丹抗缺血性中风靶点82个。GO富集分析发现,活络效灵丹治疗脑缺血性中风的靶点主要参与炎症反应、脂多糖反应、细胞因子产生的正调节、激素反应和细胞迁移的正调节等生物过程。KEGG分析提示,活络效灵丹治疗缺血性中风的作用机制与癌症通路、磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)和白细胞介素(IL)-17信号通路等有关。蛋白质互作网络和药物-活性成分-靶基因网络得到C-C基序趋化因子配体2(CCL2)、IL-1β和基质金属蛋白酶9(MMP9)等核心靶点和槲皮素、丹参酮ⅡA、白藜芦醇、姜黄素、丹酚酸、阿魏酸等10种核心成分。Pearson相关性分析显示,IL-1β和MMP9与中性粒细胞、内皮细胞等免疫细胞浸润水平相关。分子对接结果显示,IL-1β和MMP9与活络效灵丹核心成分结合稳定。结论:活络效灵丹中槲皮素、丹参酮ⅡA、白藜芦醇、姜黄素、丹酚酸和阿魏酸等有效成分能够作用于IL-1β和MMP9等核心靶点,通过调控癌症通路、PI3K/Akt和IL-17等信号通路治疗缺血性中风。
Objective:To explore the mechanism of Huoluo Xiaoling Pillets in treating ischemic stroke by using network pharmacology methods.Methods:The active ingredients and effect targets of Huoluo Xiaoling Pillets were retrieved by using Symptom Mapping(SymMap),Traditional Chinese Medicine Integrated Database(TCM-ID),Herbal Ingredient and Target Database(HIT),and Swiss Target Prediction database;the disease targets and differentially expressed genes in ischemic stroke were screened through databases such as Human Gene Database(GeneCards),Comparative Toxicogenomics Database(CTD),DisGeNET,and Gene Expression Omnibus(GEO);the effect targets of Huoluo Xiaoling Pillets,differentially expressed genes,and disease targets of ischemic stroke were intersected by using UpSetR,and the targets related to the treatment of ischemic stroke with Huoluo Xiaoling Pillets were screened out;the enrichment analysis of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were performed on relevant targets through the Metascape database.The protein-protein interaction networks and drug-active ingredient-target gene networks were constructed by Cytoscape 3.9.10 software,and the core components and targets of Huoluo Xiaoling Pillets for treating ischemic stroke were screened.The differences in immune cell infiltration between normal and ischemic samples in the GSE16561 dataset were analyzed through single sample gene set enrichment analysis,and the correlation between core targets and immune cell infiltration levels were analyzed.Molecular docking was applied to verify the affinity between the core targets and the active ingredients.Results:A total of 226 active ingredients of Huoluo Xiaoling Pillets were obtained,with 1541 targets,712 targets for ischemic stroke,and 3136 differentially expressed genes.After taking the intersection,a total of 82 anti-ischemic stroke targets were obtained for Huoluo Xiaoling Pillets.GO enrichment analysis found that the targets of Huoluo Xiaoling Pillets in treating cerebral ischemic stroke mainly participated in biological processes such as inflammatory responses,lipopolysaccharide responses,positive regulation of cytokine production,hormone responses,and positive regulation of cell migration.KEGG analysis suggested that the mechanism of of Huoluo Xiaoling Pillets in treating ischemic stroke is related to the cancer pathway,phosphatidylinositol 3 kinase/protein kinase B(PI3K/Akt),and interleukin(IL)-17 signaling pathway.The protein-protein interaction network and drug-active ingredient-target gene network yielded core targets such as C-C motif ligand 2(CCL2),IL-1β,and matrix metalloproteinase 9(MMP9),as well as 10 core components including quercetin,tanshinone IIA,resveratrol,curcumin,salvianolic acid,and ferulic acid.Pearson correlation analysis showed that IL-1βand MMP9 are correlated with immune cell infiltration levels such as neutrophils and endothelial cells.The molecular docking results showed that IL-1βand MMP9 bind stably with the core components of Huoluo Xiaoling Pillets.Conclusion:The active ingredients in Huoluo Xiaoling Pillets,such as quercetin,tanshinone IIA,resveratrol,curcumin,salvianolic acid,and ferulic acid,can act on core targets such as IL-1βand MMP9,and treat ischemic stroke by regulating the cancer pathway,and PI3K/Akt,and IL-17 signaling pathways.
作者
沈佳豪
SHEN Jiahao(Department of Pharmacy,The First People's Hospital of Huzhou,Huzhou Zhejiang 313000,China)
出处
《新中医》
CAS
2024年第13期39-47,共9页
New Chinese Medicine
关键词
缺血性中风
活络效灵丹
网络药理学
分子对接
作用机制
Ischemic stroke
Huoluo Xiaoling Pillets
Network pharmacology
Molecular docking
Mechanism
