Objective:To explore the mechanism of action of asperosaponinⅥ(AⅥ)in the treatment of rheumatoid arthritis(RA)and validate it in ex vivo experiments using network pharmacology and molecular docking methods.Methods:T...Objective:To explore the mechanism of action of asperosaponinⅥ(AⅥ)in the treatment of rheumatoid arthritis(RA)and validate it in ex vivo experiments using network pharmacology and molecular docking methods.Methods:The predicted targets of AⅥwere obtained from PharmMaper,UniProt and Swiss Target Prediction platforms,the disease targets were collected from Online Mendelian Inheritance in Man,Therapeutic Target Database and Gene Cards databases,the intersection targets of AⅥand RA were obtained from Venny 2.1.0,and the protein-protein interaction(PPI)network was obtained from STRING database,which was analyzed by Cytoscape software and screened to obtain the core targets.Cytoscape software was used to analyze PPI network and screen the core targets.Based on the Database for Annotation,Visualization and Integrated Discovery database,Gene Ontology functional and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed,and Cytoscape software was used to construct the"DiseasePathway-Target-Drug"network,which was finally verified by molecular docking and animal experiments.Results:Network pharmacological studies showed that AⅥwas able to modulate 289 targets,with 102 targets for the potential treatment of RA,with the core pathway being the AKT/PI3K signaling pathway,and the core targets being the epidermal growth factor receptor(EGFR)and matrix metalloproteinase 9(MMP9).Molecular docking results showed that AⅥcould produce strong binding with both of the 2 core targets.In vitro cellular experiments showed that AⅥreduced nitric oxide,prostaglandin E_2,tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),and IL-1βlevels(P<0.05)and inhibited cyclooxygenase-2,nitric oxide synthase,EGFR,MMP9,phosphorylated phosphoinositide 3-kinase(p-PI3K),and phosphorylated serine-threonine kinase(p-AKT)proteins(P<0.05).The results of in vivo studies showed that AⅥimproved RA score and foot swelling thickness and decreased TNF-α,IL-6,p-PI3K and p-AKT levels in RA rats(P<0.05).Conclusion:AⅥexerts anti-inflammatory and anti-RA effects which might be related to the EGFR/MMP9/AKT/PI3K pathway.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.82160779 and 82274178)the 2022 Guizhou Provincial Health Commission Science and Technology Fund Project(No.gzwkj2022-055)Guizhou Science and Technology Plan Project[No.qian ke he ji chu-ZK(2022)Yi ban 477]。
文摘Objective:To explore the mechanism of action of asperosaponinⅥ(AⅥ)in the treatment of rheumatoid arthritis(RA)and validate it in ex vivo experiments using network pharmacology and molecular docking methods.Methods:The predicted targets of AⅥwere obtained from PharmMaper,UniProt and Swiss Target Prediction platforms,the disease targets were collected from Online Mendelian Inheritance in Man,Therapeutic Target Database and Gene Cards databases,the intersection targets of AⅥand RA were obtained from Venny 2.1.0,and the protein-protein interaction(PPI)network was obtained from STRING database,which was analyzed by Cytoscape software and screened to obtain the core targets.Cytoscape software was used to analyze PPI network and screen the core targets.Based on the Database for Annotation,Visualization and Integrated Discovery database,Gene Ontology functional and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed,and Cytoscape software was used to construct the"DiseasePathway-Target-Drug"network,which was finally verified by molecular docking and animal experiments.Results:Network pharmacological studies showed that AⅥwas able to modulate 289 targets,with 102 targets for the potential treatment of RA,with the core pathway being the AKT/PI3K signaling pathway,and the core targets being the epidermal growth factor receptor(EGFR)and matrix metalloproteinase 9(MMP9).Molecular docking results showed that AⅥcould produce strong binding with both of the 2 core targets.In vitro cellular experiments showed that AⅥreduced nitric oxide,prostaglandin E_2,tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),and IL-1βlevels(P<0.05)and inhibited cyclooxygenase-2,nitric oxide synthase,EGFR,MMP9,phosphorylated phosphoinositide 3-kinase(p-PI3K),and phosphorylated serine-threonine kinase(p-AKT)proteins(P<0.05).The results of in vivo studies showed that AⅥimproved RA score and foot swelling thickness and decreased TNF-α,IL-6,p-PI3K and p-AKT levels in RA rats(P<0.05).Conclusion:AⅥexerts anti-inflammatory and anti-RA effects which might be related to the EGFR/MMP9/AKT/PI3K pathway.
