参黛清肠汤调控泛凋亡治疗溃疡性结肠炎的机制研究OA

Objective:To explore the mechanism of Shendai Qingchang(参黛清肠)Decoction in treating ulcera-tive colitis(UC)by regulating PANoptosis based on bioinformatics and animal experiments.Methods:The UC microar-ray dataset was extracted from the Gene Expression Omnibus(GEO),and the differentially expressed genes(DEGs)were obtained.The PANoptosis genes were extracted from the Genecards database,the common genes(PAN-DEGs)were extracted via the Venn diagram.The Metascape database was used for the Gene Ontology(GO)and Kyoto Ency-clopedia of Genes and Genomes(KEGG)enrichment analyses.The protein-protein interaction(PPI)network was con-structed via the STRING database.The least absolute shrinkage and selection operator(LASSO)prediction model was constructed through machine learning,and the core PAN-DEGs with diagnostic and therapeutic potential were obtained by receiver operating characteristics(ROC).The immunoinfiltration analysis was carried out with the deconvolution method(CIBERSORT).The rat model of UC was established by enema with trinitrobenzene sulfonic acid(TNBS)-ethanol.Af-ter the experiment,rat blood and intestinal tissue samples were collected as specimens,and the pathological changes of rat colon in each group were observed by hematoxylin-eosin(HE)staining.The serum levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and transforming growth factor-β(TGF-β)were quantified by enzyme-linked immunosorbent assay(ELISA),and immunohistochemistry(IHC)was used to detect the expression levels of matrix metalloproteinase 9(MMP9),interleukin 1B(IL-1β),intercellular adhesion molecule 1(ICAM1),and cluster of differentiation 44(CD44)in the colon tissue.Immunofluorescence(IF)was employed to detect the expression of zonula occludens-1(ZO-1)and mucin-2(MUC2)in the colon tissue,and the levels of diamine oxidase(DAO)and D-lactate(D-LA)were measured by corresponding assay kits.Real-time PCR was adopted to determine the mRNA levels of Mmp9,Il1b,Icam1,and Cd44 in the colon of each group,and Western blot was employed to determine the protein levels of MMP9,IL-1B,ICAM1,and CD44.Results:Datasets GSE87473 and GSE87466 were extracted from the GEO database,and 93 PAN-DEGs were obtained.KEGG analysis indicated that PAN-DEGs were mainly enriched in the nuclear factor(NF)-κB,phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT),T cell receptor,and IL-17 signaling pathways.Through PPI,LASSO regression,and ROC curve models,4 core PAN-DEGs(MMP9,IL-1B,ICAM1,and CD44)were ultimately identified.Immunoinfiltration analysis showed that the expression of core PAN-DEGs was positively correlated with activated memory CD4+T cells,follicular helper T cells,M1 macrophages,activated mast cells,and neutrophils.An-imal experiments demonstrated that compared with the normal control group,the model control group exhibited mucosal epithelial cell disarrangement,crypt deformation,and extensive inflammatory cell infiltration.In addition,the model con-trol group showed elevated serum levels of IL-6 and TNF-α(P<0.01),a declined TGF-β level(P<0.01),increased positive expression areas of MMP9,IL-1B,ICAM1,and CD44(P<0.01),weakened fluorescence intensity of ZO-1 and MUC2(P<0.01),raised levels of DAO and D-LA(P<0.01),and up-regulated mRNA and protein levels of Mmp9,Il1b,Icam1,and Cd44(P<0.01).The treatment with Shendai Qingchang Decoction at different doses alleviated the pathological changes.Particularly,the 12.96 g/kg group showed the most significant alleviating effects,with tightly ar-ranged mucosal epithelial cells,most crypt structures being clear,normal colon morphology,and only mild inflammatory infiltration in the mucosal and lamina propria layers.Moreover,this group showed declined serum levels of IL-6 and TNF-α(P<0.01),a raised TGF-β level(P<0.01),reduced staining intensity and areas of MMP9,IL-1B,ICAM1,and CD44(P<0.01),enhanced fluorescence intensity of ZO-1 and MUC2(P<0.01),lowered DAO and D-LA levels(P<0.01),and down-regulated mRNA and protein levels of Mmp9,Il1b,Icam1,and Cd44(P<0.01).Conclusion:MMP9,IL-1B,ICAM1,and CD44 are the core PAN-DEGs with diagnostic and therapeutic potential and closely related to the pathogenesis of UC.They may synergize with cells such as activated memory CD4+T cells,follicular helper T cells,and M1 macrophages to mediate the pathogenesis of UC.Shendai Qingchang Decoction may regulate the core PAN-DEGs and immune homeostasis on UC rats through NF-κB,PI3K-AKT,and T cell receptor signaling pathways,thereby relieving in-testinal mucosal injury and treat UC.