基于“提壶揭盖法”的加味真武汤治疗慢性肺心病心力衰竭作用机制的网络药理学及动物实验研究  

Network Pharmacology and Animal Experiments Reveal the Mechanism of Modified Zhenwu Decoction in Treating Chronic Pulmonary Heart Disease with Heart Failure Based on the Method of“Lifting Pot and Removing Lid”

作　　者：袁雪金 沈益 霍会爱 杨颖[2] 余清华 YUAN Xue-jin;SHEN Yi;HUO Hui-ai;YANG Ying;YU Qing-hua(Wujiang Hospital of Traditional Chinese Medicine,Suzhou Jiangsu 215221;The People′s Hospital of SND,Suzhou Jiangsu 215001;Suzhou High-Tech Zone Hospital of Traditional Chinese Medicine,Suzhou Jiangsu 215001)

机构地区：[1]苏州市吴江区中医医院,江苏苏州215221 [2]苏州高新区人民医院,江苏苏州215001 [3]苏州高新区中医医院,江苏苏州215001

出　　处：《世界中西医结合杂志》2024年第12期2414-2426,共13页World Journal of Integrated Traditional and Western Medicine

基　　金：国家重点研发计划(2018YFC2002300);苏州市第三批吴门医派名老中医药专家传承工作室项目(苏卫健中医[2023]32号)。

摘　　要：目的通过网络药理学及动物实验探讨基于“提壶揭盖法”的加味真武汤治疗慢性肺心病(Chronic pulmonary heart disease,CPHD)心力衰竭的主要信号通路及潜在作用靶点。方法通过中药系统药理学分析平台(Traditional chinese medicine systems pharmacology database and analysis platform,TCMSP)获取加味真武汤的活性成分和对应靶蛋白,Uniprot数据库获取对应靶基因;GeneCards数据库获取CPHD心力衰竭靶基因;Venny在线软件构建韦恩图,获取交集靶基因;采用STRING数据库构建蛋白互作(Protein protein interaction,PPI)网络,获得加味真武汤治疗CPHD心力衰竭的潜在核心靶基因,Cytoscape软件对结果进行可视化;Metascape数据库对潜在核心靶基因进行基因本体论(Gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析,获取加味真武汤治疗CPHD心力衰竭的主要信号通路,微生信在线软件将结果进行可视化。采用腹腔注射野百合碱溶液方法构建右心力衰竭大鼠模型,留取大鼠外周血及心脏标本。采用ELISA检测右心力衰竭大鼠外周血中TNF-a、MMP9、MMP2的含量,Western Blot、免疫组化检测右心力衰竭大鼠右心组织中ERK、p-ERK、c-Fos、c-Jun、p-c-Jun蛋白的表达水平;采用HE染色行病理检查观察右心力衰竭大鼠右心组织结构改变。采用IBM SPSS 26.0软件进行统计分析。结果通过网络药理学筛选获得加味真武汤药物有效活性成分53个,其中潜在核心有效活性成分29个;药物靶基因238个,疾病靶基因7412个,药物、疾病共有靶基因227个,其中潜在核心靶基因51个。GO分析获得1381条生物进程(Biological process,BP)、36条细胞组分(Cellular com⁃ponent,CC)、211条分子功能(Molecular function,MF);KEGG通路富集分析获得PI3K/AKT、MAPK、IL-17、TNF等共262条信号通路。动物实验结果显示加味真武汤可调控ERK/AP-1信号通路活性,抑制模型大鼠外周血中MMP-2Objective To investigate the main signaling pathways and potential targets of modified Zhenwu Decoction in treating chronic pulmonary heart disease(CPHD)with heart failure through network pharmacology and animal experiments.Methods Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)was searched for the active ingredients and corresponding target proteins of modified Zhenwu Decoction.The corresponding target genes were obtained from Uniprot.The target genes of CPHD with heart failure were obtained from GeneCards.Venny online was used to establish the Venn diagram to obtain common target genes.STRING was employed to construct the protein-protein interaction(PPI)network,and the potential core target genes of modified Zhenwu Decoction in treating CPHD with heart failure were obtained.The results were visualized by Cytoscape.Metascape was used for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses of the potential core target genes,on the basis of which the main signaling pathways of modified Zhenwu Decoction in treating CPHD with heart failure were obtained.The results were visualized by Wei Sheng Xin online.The rat model of right heart failure was established by intraperitoneal injection of monocrotaline solution,and the peripheral blood and heart samples were collected.The levels of tumor necrosis factor-alpha(TNF-α),matrix metallopeptidase(MMP)9,and MMP2 in the peripheral blood were measured by enzyme-linked immunosorbent assay(ELISA),and the expression levels of extracellular signal-regulated kinase(ERK),phosphorylated ERK(p-ERK),c-Fos,c-Jun,and p-c-Jun in the right heart tissue of the rat model of right heart failure were determined by Western blotting and immunohistochemistry.Hematoxylin-eosin staining was used to observe the structural changes of the right heart tissue in the rat model of right heart failure.IBM SPSS 26.0 was used for statistical analysis.Results A total of 53 active ingredients of modified Zhenwu Decoction were retrieved,among which

关 键 词：加味真武汤 慢性肺心病心力衰竭 网络药理学 提壶揭盖法 ERK/AP-1信号通路 

分 类 号：R285.6[医药卫生—中药学]