lncRNA-miRNA-mRNA Network of Yuye Decoction(玉液汤)for the Prevention and Treatment of Diabetic Nephropathy Based on GEO Database
Objective:To excavate the ternary transcriptional network of Yuye Decoction(YYD,玉液汤)against diabetic nephropathy(DN),and to predict its biomarkers and mechanism of action in the prevention and treatment of DN.Methods:Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Pharmmapper,and Uniprot were used to predict the key active components of YYD and their target proteins,and through GEO database,differentially expressed mRNA and miRNA in normal and DN patients were screened.The predicted targets of YYD were intersected with differentially expressed mRNAs(DEmRNAs)in DN to obtain the targets of YYD against DN.String was used to map the protein-protein interaction(PPI)network,and Metascape was utilized for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The active component-target network of YYD in the prevention and treat-ment of DN was constructed by Cytoscape 3.6.1.The targets of YYD in the PPI network were imported into Cytoscape for topological analy-sis,and the core targets were screened using the Cytohubba plug-in.The miRDB,miRWalk,and TargetScan databases were used to predict miRNAs associated with the core targets,which were intersected with the DEmiRNAs in DN.The shared miRNAs were then imported into the Starbase database to predict the related lncRNAs,and DN-related lncRNAs were screened.Discovery Studio 2019 was used to perform the molecular docking of core compounds-core targets.Streptozotocin(STZ)combined with high-fat and high-sugar feed was given to induce DN rat model.The rats were treated with YYD 2.6 g/kg to observe the changes in body mass,fasting blood glucose,and histopathological mor-phology of the kidneys in each group.Additionally,urinary protein,blood creatinine,and urea nitrogen were detected.Fluorescence quantita-tive polymerase chain reaction(Real-time PCR)was performed to validate the differential genes,and the competitive endogenous RNA(ceR-NA)network of lncRNA-miRNA-mRNA of YYD against DN was established by the Cytoscape.Results:The ceRNA network was composed of 17 key lncRNAs,10 key miRNAs and 13 key mRNAs.Experiments verified that in the ceRNA network,compared with the conditions in the model control group,urinary protein,blood creatinine and urea nitrogen were significantly reduced in the YYD 2.6 g/kg group,and the ln-cRNA expressions of Neat 1,Xist,and Malat1,as well as the mRNA expressions of Casp1,Ctsb,and Ctsd were significantly down-regulated,while the miRNA expressions of miR-361-3p,miR-193a-3p,and miR-139-5p were markedly up-regulated.GO and KEGG analyses revealed that the key targets involved in biological processes such as inflammation,fibrosis,oxidative stress,apoptosis,and autophagy in the kidney mainly through the modulation of apoptosis,NOD-like receptor signaling pathway,MAPK signaling pathway,IL-17 signalling pathway,TGF-be-ta signaling pathway and PI3K-Akt signaling pathway and other pathways.Molecular docking showed that the active components,folic acid,calycosin,quercetin and Icariin I were well bound to the core targets.Conclusion:YYD may protect the kidney and inhibit renal inflammation by affecting the lncRNA-miRNA-mRNA network in DN rats,which involves multi-component,multi-target and multi-pathway synergistic effects.
万方数据
维普
