| Objective: Predict the key receptor proteins and signaling pathways of Anwei Decoction on chronic atrophic gastritis(CAG)based on the principle of network pharmacology,and explore the influence of Anwei Decoction on the key factors of the signaling pathway through experimental verification,combined with molecular docking for further research combination types of active ingredients and key factors,and in-depth discussion of the effect of Anwei Decoction on the EGFR-AKT-GSK3β signaling pathway in chronic atrophic gastritis.Methods: The network pharmacology method was used to construct the network relationship between the effective components of Anwei Decoction and its targets,and the pathway enrichment analysis was carried out;the CAG rat model was replicated with sodium deoxycholate and ammonia water,and the epidermal growth factor receptor(EGFR)was detected after the intervention of Anwei Decoction,protein kinase B(AKT),glycogen synthase kinase-3β(GSK3β)relative expression in gastric tissue;The effective monomer component is docked with the protein simulation,and the effective component of the drug is predicted and visualized according to the size of the binding energy.Results: There are 28 common targets at the intersection of Anwei Decoction drug component targets and CAG-related genes,among which AKT has the highest degree of correlation(Degree),KEGG pathway enrichment screening shows that it is involved in cancer pathways,EGFR tyrosine kinase inhibitors,Helicobacter pylorus Epithelial cell signaling,gastric cancer pathway and other signaling pathways in bacterial infection;EGFR,AKT,and GSK3β may be key factors in the treatment of CAG.Animal experiments with hematoxylin-eosin(HE)staining showed that the structure of each layer of the gastric mucosa of rats in the normal control group was intact,and the lamina propria of the gastric mucosa in the model control group showed inflammatory cell infiltration.It can be seen that the infiltration of chronic inflammatory cells decreased,and the atrophy was significantly improved.Western blot(WB)and real-time fluorescent quantitative PCR(q RT-PCR)were used to observe the gastric tissue of CAG rats.Compared with the normal control group,the expressions of AKT and EGFR in the model control group were significantly up-regulated(P <0.05),the expression of GSK3β was significantly down-regulated(P<0.05);compared with the model control group,the high-dose Anwei Decoction group could significantly reduce the expression of AKT and EGFR(P<0.05)and increase the expression of GSK3β(P<0.05).Molecular docking showed that there are 9 small molecule ligands in Anwei Decoction that can tightly combine with different sites of receptor proteins to form stable chemical bonds such as hydrogen bonds,cationic πbonds,and π-π stacking.Among them,isorhamnetin showed excellent binding ability;in addition,luteolin,kaempferol and quercitrin could all bind well to the target receptor protein;finally,five compounds including baicalin had different degree of integration.In conclusion,isorhamnetin,luteolin,kaempferol and quercitrin may be the effective monomer components of Anwei Decoction in the treatment of CAG.Conclusion:(1)Network pharmacology predicts that the treatment of CAG by Anwei Decoction may be related to the expression of the EGFR-AKT-GSK3βsignaling pathway,and the key targets may be EGFR,AKT and GSK3β.(2)Animal experiments have verified that Anwei Decoction can improve the pathological morphology of gastric mucosa in CAG rats,and at the same time significantly down-regulate the expression of AKT and EGFR and upregulate the expression of GSK3β.(3)Molecular docking showed that the effective monomer components of Anwei Decoction in the treatment of CAG may be isorhamnetin,luteolin,kaempferol and quercetin. |
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