Potential pathogenic mechanism of ciprofloxacin in regulating aortic dissection based on network pharmacology and experimental verification
Objective To screen the key target genes of ciprofloxacin in regulation of the occurrence and development of aortic dissection through network pharmacology and molecular docking technology,and further explore its potential pathogenic mechanism through animal and cell experiments.Methods TO screen the related targets of ciprofloxacin and aortic dissection were from TargetPrediction,PharmMapper,GeneCards,and CTD databases respectively,and the potential targets of ciprofloxacin regulating aortic dissection were obtained after the intersection of the related targets.PPI network was constructed using the search tool for STRING database to analyze the association between potential targets.The potential targets were analyzed by KEGG pathway enrichment and GO function enrichment analysis,and the"drug-target-pathway-disease"network was constructed.Cytoscape software and MCODE and other plug-ins were used to further screen out the key targets.The chemical structure of ciprofloxacin and the key targets were docked and visualized by AutoDock vina and PyMol software.Combined with animal model construction,HE staining,cell proliferation,and toxicity detection experiments,RT-Qpcr experiments,and Western blotting were used to detect the key target gene expression in mouse aortic tissues and human aortic smooth muscle cells(HASMCs).Results In this study,515 Ciprofloxacin-related targets,and 9 004 aortic dissection-related targets were screened,resulting in 412 potential ciprofloxacin-regulated aortic dissection targets.KEGG pathway enrichment analysis showed that they were mainly enriched in lipid and atherosclerosis,IL-17 signaling pathway,TNF signaling pathway,etc.GO functional enrichment analysis showed that CC was mainly enriched in membrane raft and membrane microdomain.MF was mainly enriched in cytokine receptor binding and xenobiotic transmembrane transporter activity.BP were mainly enriched in the response to lipopolysaccharide,the response to molecule bacterial origin,etc.STRING and Cytoscape analysis were used to construct the PPI network and key sub-modules,and 15 key targets were obtained.They were Bcl-2,JUN,CD44,CASP3,TLR4,IFNG,TP53,Akt1,ALB,MMP9,IL-6,IL10,IL-1B,GAPDH,and TNF.Molecular docking results showed that ciprofloxacin had a good docking mode with MMP9,ALB,GAPDH,Akt1,TP53,CASP3,and IL1β.In vivo experiments showed that compared with the model group,the model+ciprofloxacin model group had increased film formation rate and mortality rate,and more serious damage to the elastic structure of the vessel wall.The mRNA and protein expression levels of pro-apoptotic and pro-inflammatory genes were increased.In vitro experiments showed that compared with the control group,the expression levels of MMP9,IL-6,CASP3,JUN,IL-1β,TP53,and TLR4 were significantly up-regulated,and the expression level of Akt1 was significantly down-regulated in HASMCs cell stimulated with different concentrations of ciprofloxacin.Conclusion Ciprofloxacin is one of the important factors mediating the occurrence and development of aortic dissection,which may play a regulatory role by stimulating the expression of inflammatory factors and VASMCs cell apoptosis.
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