The Mechanism of Action of Zhigangao Decoction for the Treatment of Arrhythmia Based on Network Pharmacology,Molecular Docking and Animal Experiments
GUO Changxue
GAO Yang
ZHAO Xiao
QIN Hewei
Abstract：Objective:Based on network pharmacology,molecular docking technology and combined with animal experiments to verify and analyze the mechanism of action of Zhigangao Decoction for the treatment of arrhythmia.Methods:The active components and their targets of zhigancao,ginseng,cassia twig,Ziziphus jujuba seed,Ziziphus jujuba mill,Shengdi and e Jiao were obtained through the TCSMP database.Gene Cards database was used to obtain the targets related to arrhythmias.Venn diagram was used to obtain the common targets.The protein-protein interactions(PPI)were analyzed through the STRING database,the PPI network graph was constructed,and the core targets were screened through network topology analysis.Gene ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were conducted on the core targets.The Cytoscape 3.9.1 software was used to draw the multi-dimensional correlation visualization network of drugs-active ingredients-targets of Zhigangao Decoction,and the key active ingredients were obtained through network analysis.The key active ingredients and targets screened out were verified for molecular docking using AutoDock.Arrhythmia was induced by injecting aconitine into the left external jugular vein.After successful modeling,the intervention was carried out with Zhigancao Decoction.After 15 consecutive days of intervention,the electrocardiograms of rats were recorded using a biological experimental functional system.The were recarded incidence and duration of ventricular tachycardia,as well as ventricular fibrillation.The core target levels in the blood and aorta of rats were detected by enzyme-linked immunosorbent assay(ELISA),real-time fluorescence quantitative polymerase chain reaction(RT-PCR),and Western Blot.Results:A total of 165 chemical components were analyzed,among which quercetin,kaempferol,β-sitosterol,stigmasterol and fumarine were five key active components.After de-duplication of the predicted targets,392 targets were obtained.Intersection mapping was conducted with 1,228 disease-related targets,and 95 targets were obtained.The protein interaction network screened out 20 core targets including interleukin-6(IL6),protein kinase B1(AKT1),tumor necrosis factor(TNF),interleukin-1 β(IL1B),cyclooxygenase 2(PTGS2),hypoxia-inducible factor 1(HIF-1),and matrix metalloproteinase 9(MMP9).GO functional enrichment analysis included 647 biological process items,74 cellular component items,and 88 molecular function items.KEGG enriched 167 pathways.The molecular docking results showed that five key compounds and six core caned interact directly.The results of animal experiments showed that Zhigancao Decoction could significantly improve the conditions of arrhythmia(latency time,maintenance time,heart rate,and incidence of arrhythmia)in rats(P＜0.05),and reduce the levels of AKT1,TNF,and IL1B in the peripheral serum of rats(P＜0.05),the mRNA and protein levels of HIF-1α,PTGS2 and MMP9 in rat ventricular tissues reduced(P＜0.05).Conclusion:The Zhigancao Decoction could improve arrhythmia and its mechenism may be related to regulating of HIF-1A pathway to inhibit the expression of key targets such as HIF-1α,PTGS2,MMP9,AKT1,TNF,and IL1B.
Keywords：arrhythmiaZhigancao Decoctionnetwork pharmacologymolecular dockingexperimental verification
Publication Date：2025-06-10
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：11( 1625-1635 )