Mechanism Of Cangzhu In Treating Gastric Ulcer Based On Network Pharmacology

Objective.To analyze the active ingredients,core targets,and mechanism of action of Cangzhu in the treatment of gastric ulcer(Gastric ulcer,GU)using a network pharmacology approach,and to verify the core targets by animal experiments.Methods.Ⅰ.Network pharmacology component.1.The chemical composition of Cangzhu was obtained by searching the three databases of TCMSP,TCMID,and Chinese Natural Products Chemical Composition Database,and the potential active ingredients of Cangzhu were obtained by screening the chemical composition using OB and DL values,and finally,they were supplemented with the literature.These active ingredients were searched through the TCMSP database,STITCH database,Swiss Target Prediction database to obtain their corresponding direct targets,and the abbreviations corresponding to the targets were obtained through the Uniprot database.The STRING database was then used to expand these targets,and the two were aggregated to obtain the relevant targets of Cangzhu.2.Search the Genecards,Drug Bank,and TTD databases to find the targets related to gastric ulcer,and expand the above targets by STRING database to obtain the gastric ulcerrelated targets.3.The potential targets of Cangzhu for the treatment of gastric ulcer were obtained by intersecting Cangzhu-related targets and finding the active ingredients and GU-related targets upstream and downstream of the potential targets,and constructing the "ingredient-targetdisease" network by using Cytoscape 3.8.1 software,performing topological analysis on the network,and using The network was then topologically analyzed and the core targets and core components for the treatment of GU were obtained using mean screening.4.Use DAVAD 6.8 GO analysis and KEGG pathway analysis to enrich the biological pathways involved in the core targets.5.The core targets were screened using the scores of BC,CC,DC,and genecard from the topological analysis.The components corresponding to the core targets,the core components screened by the topological analysis were summarized and validated with the core targets by molecular docking with Maestro 11.9 software.Ⅱ.Animal experiment part.1.Establishment of a rat-bound-water immersion gastric ulcer model.2、To observe the effect of Cangzhu on ulcer index(UI)in rats.3.HE staining to observe the cell morphology of gastric tissues.4.ELISA to detect the expression levels of three core target proteins(AKT1,PTGS2,VEGFA)in gastric tissues.Results.Ⅰ.Cyberpharmacology section.1.27 active ingredients of Beicangzhu were obtained by searching,of which 21 active ingredients had eligible targets;43 active ingredients of Nancangzhu had eligible targets,of which 33 active ingredients had eligible targets.A total of 1026 GU targets were obtained.2.The "ingredient-target-disease" network of Beicangzhu was constructed,containing 21 active ingredients and 945 targets,and 177 core targets were obtained by topological analysis.3.Comparing the core components and the top ten core targets of Beicangzhu and Nancangzhu,we found that five core components were the same(wogonin,atractylone,alphaeudesmol,hinesol,beta-eudesmol),in addition to Ar-curcumene,3β-acetoxyatractylone,5-methylolfurfural,and Diisobutyl phthalate were unique to Nancangzhu,and limonin,Oleic acid,Aristolone,and Furanodiene were unique to Beicangzhu.8 of the top 10 core targets are the same for both(TP53,TNF,IL6,AKT1,PTGS2,CXCL8,CDKN1 A,IL1B).Also,TERT and CCND1 are unique to Nancangzhu,and VEGFA and SST are unique to Beicangzhu.This indicates that the components and targets that play a dominant role in the treatment of gastric ulcers are highly similar between the two.4.The results of GO functional analysis enrichment showed that there were 485 enrichment pathways in Nancangzhu and 566 in Beicangzhu,and 425 intersecting pathways in both,accounting for 87.63% and 75.09% of the total enrichment pathways in Beicangzhu and Nancangzhu,respectively.The enrichment results of KEGG pathways showed that 126 pathways were obtained from Nancangzhu and 134 from Beicangzhu.The two pathways intersected 123 pathways,accounting for 97.62% and 91.79% of the total number of pathways in Beicangzhu and Nancangzhu,respectively.Based on the function of the pathways,it is speculated that Cangzhu achieves the treatment of gastric ulcer through anti-inflammatory,antibacterial,anti-cancer,anti-oxidation,and regulation of cell growth and apoptosis.5.The molecular docking scores of the core components of Beicangzhu were all less than0,and the average docking score of each core target was less than-4 except for IL1 B and TNF,indicating that the core components could act well on the core targets.Ⅱ.Experimental part1.Ulcer index(UI): compared with the blank group,the UI of the model group was significantly higher(P < 0.05);compared with the model group,the UI of each administration group was significantly lower(P < 0.05),and the UI of the high,medium and low dose groups of Cangzhu increased in a gradient,and the P-value between the groups was less than 0.05.2.Morphological observation: The gastric tissues of the rats in the blank group were normal and no cell damage was seen.while the gastric tissues of the rats in the model group showed necrosis and detachment of mucosal cells,with local involvement of the whole layer of the mucosa and congestion and edema in the blood vessels of the submucosa.The most significant reduction was seen in the high dose group and the positive control group.3.The expression levels of AKT1 and PTGS2 in the gastric tissues of rats with boundflooded gastric ulcers were reduced and the expression level of VEGFA was increased.Conclusion1.Cangzhu can participate in the therapeutic process of gastric ulcer with multiple targets and pathways.The core components of Cangzhu,such as Atractylodin and atractylone,can inhibit the inflammatory response,promote gastric mucosal tissue repair and inhibit excessive cell proliferation through regulating the core targets of AKT1,PTGS2,VEGFA,IL6,and TNF to achieve the purpose of anti-gastric ulcer.2.Cangzhu had a protective effect on atherogenesis in a rat model of bound-flooded gastric ulcer and was able to reduce the expression levels of AKT1 and PTGS2(COX-2)and increase the expression level of VEGFA in their gastric tissues.