基于网络药理学和体外实验的异绿原酸B治疗炎症性肠病作用机制研究OA

Objective To explore the therapeutic targets and mechanisms of Isochloroic acid B(ICAB)in the treatment of inflammatory bowel disease(IBD)by network pharmacology,molecular docking and in vitro experiments,providing new ideas and scientific evidence for the potential clinical application of ICAB in IBD and the subsequent development of drugs for the treatment of IBD.Methods Databases including SwissTargetPrediction,PharmMapper,and GeneCards were used to predict and screen the common targets of ICAB and IBD.Gene ontology(GO)functional and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analyses were performed to screen the top 5 core targets and the top 10 signaling pathways.Molecular docking was conducted to validate the interaction between ICAB and the core targets.Concurrently,an in vitro inflammatory injury model was established by 40 μg/mL lipopolysaccharide(LPS)to induce human colonic epithelial cells(NCM460),in order to validate the anti-inflammatory effects and mechanism of action of ICAB.Results A total of 287 component targets of action,2 525 disease-related targets,and 105 targets of action common to ICAB and IBD were predicted.The protein-protein interaction(PPI)network topology showed that the top five key targets were ALB,AKT1,MMP9,GASP3,and EGFR.Molecular docking results showed that the binding energies between ICAB and the core targets were all below-29.30 kJ/mol,indicating strong binding activity.KEGG enrichment analysis indicated that the pathways were primarily concentrated on the Relaxin signaling pathway,Proteoglycans in cancer,MAPK signaling pathway,PI3K-AKT signaling pathway.In vitro cell experiments demonstrated that ICAB could reduce the levels of interleukin-8(IL-8)and interleukin-1β(IL-1β)in the LPS-induced NCM460 cell inflammatory injury model(P<0.05),decrease the mRNA expression of PI3K and AKT(P<0.05),and reduce the protein expression of p-PI3K,p-AKT,p-mTOR,as well as the ratios of p-PI3K/PI3K,p-AKT/AKT and p-mTOR/mTOR(P<0.05).Conclusion ICAB exhibits anti-inflammatory effect against IBD,and its mechanism of action is likely associated with the inhibition of the PI3K/AKT/mTOR signaling pathway.