湖南中医药大学学报2025，Vol.45Issue(12)：2296-2306,11.DOI:10.3969/j.issn.1674-070X.2025.12.008
基于网络药理学和细胞实验探讨络风宁2号方治疗心力衰竭的作用机制
Mechanism of action of Luofengning Formula No.2 in treating heart failure based on network pharmacology and cellular experiments
摘要
Abstract
Objective To explore the mechanism of action of Luofengning No.2 Formula(LFN2)in treating heart failure.Methods The active components and potential targets of LFN2 were identified using TCMSP and SwissTargetPrediction databases,while heart failure-related disease targets were collected from GeneCards and OMIM databases.Intersecting targets between LFN2 and heart failure were determined using Venny 2.1.0 online platform,then imported into STRING database to construct a protein-protein interaction(PPI)network.Cytoscape 3.9.1 software was used to visualize the"drug-component-target"network.R language was employed for GO functional enrichment analysis and KEGG pathway enrichment analysis.Molecular docking was performed to validate the binding affinity between the active components of LFN2 and the core targets.Cellular experiments were conducted to validate the cAMP/Drp1 and apoptosis pathways.Cells were divided into control,model,LFN2,Db-cAMP,and LFN2+Db-cAMP groups.The expression levels of ANP/BNP mRNA,cAMP,Drp1,and apoptosis-related proteins(cleaved Caspase-3,Bcl-2,and Bax)were measured in each group.Results Network pharmacology identified 157 active components and 553 target proteins for LFN2.KEGG pathway enrichment analysis revealed significant associations with the Wnt signaling pathway,Rap1 signaling pathway,MAPK signaling pathway,PPAR signaling pathway,cAMP signaling pathway,glucagon signaling pathway,and renin secretion pathway.PPI network analysis identified AKT1,ALB,and TNF as core targets.Molecular docking results showed that the main active components(Ginsenoside Rh4,Biatractylolide,Danshenxinkun D,and Tumulosic acid)exhibited strong binding affinities(binding energy<-5.10 kJ/mol)with the core targets(GAPDH,TNF,IL6,AKT1,and ALB).Cellular experiments demonstrated that compared with the control group,the model group showed increased ANP/BNP mRNA content(P<0.01),decreased cAMP content(P<0.05),enhanced Drp1 fluorescence intensity and protein level(P<0.05),significantly elevated levels of pro-apoptotic proteins cleaved Caspase-3 and Bax(P<0.01),and decreased levels of anti-apoptotic protein Bcl-2(P<0.05).Compared with the model group,the LFN2,Db-cAMP,and LFN2+Db-cAMP groups exhibited reduced ANP/BNP mRNA content(P<0.05),increased cAMP content(P<0.05),reduced Drp1 fluorescence intensity and protein level(P<0.05),and increased Bcl-2 protein level(P<0.05).The improvement was more pronounced in the LFN2+Db-cAMP group than in the LFN2 group.Conclusion LFN2 may exert anti-heart failure effects by regulating the cAMP/Drp1 signaling pathway and inhibiting cardiomyocyte apoptosis.关键词
心力衰竭/络风宁2号方/网络药理学/cAMP/Drp1/细胞凋亡Key words
heart failure/Luofengning Formula No.2/network pharmacology/cAMP/Drp1/apoptosis分类
医药卫生引用本文复制引用
郑清华,刘一卓,秦子镒,邱璐,郑相颖,王显..基于网络药理学和细胞实验探讨络风宁2号方治疗心力衰竭的作用机制[J].湖南中医药大学学报,2025,45(12):2296-2306,11.基金项目
北京市科研基地建设项目(2025-JYB-KYPT-05) （2025-JYB-KYPT-05）
国家自然科学基金项目(82074263). （82074263）
