基于动物实验利用网络药理学及分子对接技术探讨杜仲治疗肺动脉高压的作用机制  

Mechanism of Eucommia ulmoides for pulmonary hypertension based on animal experiments and network pharmacology combined with molecular docking

作　　者：李瑞湘 金远香 史志华 王瑾 LI Rui-xiang;JIN Yuan-xiang;SHI Zhi-hua;WANG Jin(Department of Ultrasonography,Hunan Chest Hospital,Changsha 410000;Department of Pharmacy,Hunan University of Medicine General Hospital,Huaihua Hunan 418000;Department of Pharmacy,Changsha Central Hospital,Changsha 410000)

机构地区：[1]湖南省胸科医院超声科,长沙410000 [2]湖南医药学院总医院超声科,怀化418000 [3]湖南医药学院总医院药学部,怀化418000 [4]长沙市中心医院药学部,长沙410000

出　　处：《中南药学》2024年第10期2633-2642,共10页Central South Pharmacy

基　　金：湖南省自然科学基金(No.2020JJ5445);吉首大学校级课题(No.Jdzd21035);白求恩医学科学研究基金(No.TY046BS)。

摘　　要：目的基于动物实验与网络药理学及分子对接技术探讨杜仲治疗肺动脉高压(PH)的分子机制。方法(1)构建野百合碱(MCT)诱导的PH大鼠模型,观察杜仲提取物对PH大鼠的治疗作用,利用HE染色观察肺组织的形态学变化,酶联免疫分析检测外周血浆中炎症因子HMGB1、TNF-α、IL-1β及IL-6水平。(2)基于TCMSP、GeneCards、OMIM、TTD以及DrugBank等公共数据库信息,筛选出杜仲和PH的靶点基因,进行靶点标准化;运用Cytoscape 3.9.1软件构建杜仲活性成分-PH-靶点网络;将获得的关键靶标上传到STRING 11.0数据库,用于蛋白质-蛋白质相互作用(PPI)网络分析;通过拓扑分析确定杜仲治疗PH的核心靶标,并进行GO功能富集分析和KEGG信号通路富集分析;结合分子对接技术验证杜仲治疗PH的活性成分与核心靶标之间结合活性。(3)利用免疫组织化学染色检测潜在靶点的蛋白表达水平。结果(1)动物实验结果表明,杜仲提取物可以降低MCT诱导PH大鼠的右心室收缩压(RVSP),缓解右心室肥厚及肺血管重构,降低外周血浆中HMGB1、IL-1β、IL-6及TNF-α水平。(2)网络药理学结果显示,筛选得到杜仲活性成分25种,相关靶点195个,PH相关靶点6416个。杜仲治疗PH的潜在核心靶点19个。GO功能富集分析共获得2789个条目,包括生物过程2477条、细胞组分101条及分子功能211条。KEGG信号通路富集分析,其中主要信号通路包括PI3K-Akt信号通路、IL-17信号通路及TNF信号通路等。分子对接显示,杜仲核心活性成分与靶点有较强结合能力。(3)免疫组织化学染色显示杜仲提取物明显降低了PH大鼠肺组织中AKT1、ESR1、STAT3和IL-6的表达,而增加了TP53的表达水平。结论本研究初步揭示了杜仲可通过多成分、多靶点、多途径发挥治疗PH的作用,为其临床应用和深度开发提供研究基础。Objective To determine the molecular mechanism of Eucommia ulmoides for pulmonary hypertension(PH)based on animal experiments and network pharmacology combined with molecular docking.Methods(1)The experimental PH rat model was established with a single intraperitoneal injection monocrotaline(MCT)and then the effect of extract from Eucommia ulmoides on PH rat was observed.Hematoxylin-eosin staining and enzyme linked immunosorbent assay were used to analyze the morphology of lung tissues and to detect HMGB1,TNF-α,IL-1β,and IL-6 level in the plasma.(2)Based on TCMSP,GeneCards,OMIM,TTD,and DrugBank,the target genes of Eucommia ulmoides and PH were screened and standardized.The Eucommia ulmoides ingredients-PH-target network was established with Cytoscape software.The key targets obtained were uploaded to the STRING database for protein-protein interaction(PPI)network analysis.The core targets of Eucommia ulmoides in the treatment of PH were identified by topological analysis.Additionally,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were performed.Molecular docking was used to verify the binding activity between the active ingredients of Eucommia ulmoides and the core target for PH.(3)Immunohistochemistry staining was used to detect the expression level of potential key targets.Results(1)Animal experiments show that the extract from Eucommia ulmoides relieved MCT-induced PH.Haemodynamic parameters including RVSP,RV/(LV+S),RV/(Tibial length)and the thickness of pulmonary arterial wall were greatly reduced in the PH rats.Meanwhile,the level of HMGB1,IL-1β,IL-6,and TNF-αin PH rats were reduced as well.(2)Network pharmacology showed 25 active components of Eucommia ulmoides,195 related targets,and 6416 PH-related targets.PPI network topology analysis uncovered 19 potential core targets of Eucommia ulmoides for PH treatment.A total of 2789 items were obtained via GO functional enrichment analysis,including 2477 biological processes,101 cellular components,and 211 molecular functions.Molecular docki

关 键 词：杜仲 肺动脉高压 网络药理学 分子对接 作用机制 

分 类 号：R285.5[医药卫生—中药学]