| Research Objective:Crohn’s disease(CD)is a chronic and inflammatory granulomatous disease of digestive tract with unknown etiology,and it is a kind of Inflammatory bowel disease(IBD).Its incidence is on the rise in the world,Asia and China.At present,the diagnosis and evaluation of CD depends on endoscopy and imaging examination,and the study of convenient and accurate serological indicators in clinic is a hot spot.About 75% of CD patients have intestinal lesions involving Terminal ileum(TI),and CD is also one of the common causes of TI lesions,so TI lesions may be associated with early manifestations of CD.Based on bioinformatics technology,this paper studied whether there are Differentially expressed genes(DEGs)between CD patients with TI inflammation(TICD group)and CD patients without TI inflammation(n TICD group),screened biological markers related to early diagnosis of CD,and evaluated the value of these markers in clinical diagnosis and curative effect evaluation of CD.Research Methods:1.Data source and processingIn GEO database,the CD-related data set GSE73094 was retrieved and downloaded,and analyzed by GEO2 R online tool software.According to the presence or absence of CD,it was divided into CD group and healthy control group.In the CD group,according to whether TI has inflammation,the CD group is divided into TICD group and n TICD group.DEGs;of CD group and healthy control group,TICD group and n TICD group were obtained respectively.Two groups of PPI network diagrams of DEGs were constructed respectively to screen key genes;Take the intersection of two groups of key genes as the research goal.2.Experimental objects and methods The clinical data and serum samples of patients who were hospitalized in a 3A hospital in Jiangsu Province from August 2022 to June 2023 and those who were diagnosed as CD due to TI lesions were collected.Serum MCP-1,IFN-γ and ICAM1-1 levels were detected by ELISA kit.2.Experimental objects and methodsThe clinical data and serum samples of patients hospitalized in a 3A hospital in Jiangsu Province from June 2022 to June 2023 who were diagnosed with CD,and patients who were diagnosed with TI lesions due to colonoscopy during the same period but were not diagnosed with CD were collected.Serum MCP-1,IFN-γ and ICAM1-1 levels were detected by ELISA kit.3.Experimental groupingPatients were divided into CD group and terminal ileitis group(n CD group).The CD group was divided into early diagnosis group and previous diagnosis group according to the course of disease.According to Harvey-Brashow score,they were divided into remission group and activity group;According to the imaging results of colonoscopy,abdominal CT and MRI,they were divided into terminal ileum involvement group(TICD group)and non-involvement group(n TICD group).4.Statistical methodThe general clinical data and serum MCP-1,IFN-γ and ICAM1-1 levels were compared among the groups.Spearman correlation analysis was used to compare the relationship between serum MCP-1,IFN-γ and ICAM1 levels and clinical indexes of patients.Logistic regression was used to analyze the relationship between CD disease activity and serum MCP-1,IFN-γ,ICAM1 levels and clinical indexes.ROC curve was used to evaluate the predictive value of serum MCP-1,IFN-γ and ICAM1 levels on the degree of disease activity.Research results:1.A total of 225 DEGs were screened from 1.CD group and healthy control group,including 161 up-regulated genes and 64 down-regulated genes(FDR < 0.05).A total of 371 DEGs were screened from TICD group and n TICD group,including214 up-regulated genes and 157 down-regulated genes(FDR<0.05).2.The PPI network diagram of two groups of DEGs was constructed,and the top10 genes with the highest MMC score were selected as key genes: MCP-1,CXCL1,CXCL8,ICAM1,IFN-γ,RELA,CTLA4,FOS,TNFRSF1 A and STAT1;were the key genes in CD and healthy people;The key genes in TICD group and n TICD group are CD40,CXCL8,ICAM1,ITGAX,IFN-γ,IL2,MCP-1,ITGAM,CTLA4 and IL10.Taking the intersection of two groups of key genes,a total of five genes overlap,and their expressions are consistent in the two groups,namely CXCL8,ICAM1,IFN-γ,CTLA4 and MCP-1,all of which are up-regulated genes.KEGG analysis showed that the functions of these five key genes were mainly enriched in IL-17 signaling pathway and cytokine-cytokine receptor signaling pathway.3.Clinical verification:3.1 The levels of MCP-1,IFN-γ and ICAM1-1 in TICD group were higher than those in n TICD group,P < 0.05;3.2 The levels of serum IFN-γ and ICAM1-1 in early diagnosis group were higher than those in n CD group(P < 0.01).The levels of serum MCP-1 and ICAM1-1in previous diagnosis group were higher than those in n CD group(P < 0.001).The serum MCP-1 level in the previous diagnosis group was higher than that in the early diagnosis group(P < 0.05);3.3 The levels of serum MCP-1,serum ICAM-1,IL-6 and ESR in active CD were higher than those in remission group(P < 0.01);3.4 Spearman correlation analysis showed that serum IFN-γ level was positively correlated with FC(ρ=0.38,P < 0.01),ESR(ρ=0.33,P < 0.01)and CRP(ρ=0.31,P < 0.05).Serum ICAM-1 level was positive with FC(ρ=0.41,P <0.01),IL-6(ρ=0.53,P < 0.01),ESR(ρ=0.44,P < 0.01)and CRP(ρ=0.57,P < 0.01);3.5 Binary Logistic regression analysis suggests that MCP-1,IFN-γ,ICAM-1,IL-6,ESR and CRP are independent predictors of disease activity in CD patients,that is,the higher the levels of MCP-1,IFN-γ,ICAM-1,IL-6,ESR and CRP,the higher the possibility of disease activity in CD patients;3.6.ROC analysis showed that the area under the ROC curve for MCP-1 to predict CD disease activity was 0.8244;The area under ROC curve for IFN-γ to predict CD disease activity is 0.6197: The area under ROC curve predicted by ICAM-1 is 0.8378.That is,MCP-1,IFN-γ and ICAM-1 have potential value in predicting whether CD is active or not.Research conclusions:1.DEGs exist between CD patients and healthy people,CD patients with TI involvement and CD patients without TI involvement,and DEGs are all related to the activation of inflammatory signaling pathway,indicating that the activation of inflammatory signaling pathway may play an initial trigger role in the occurrence and development of CD.2.Serum MCP-1,IFN-γ and ICAM1-1 levels have potential role in the differential diagnosis of CD;The levels of MCP-1,IFN-γ and ICAM-1 in serum have a potential role in differentiating CD from TI lesions.Serum IFN-γ and ICAM1 levels may have potential value in early diagnosis of CD.3.The levels of IL-6,ESR,CRP,MCP-1 and ICAM-1 in serum are risk factors for the disease activity of CD patients.Serum MCP-1,IFN-γ and ICAM-1 levels have potential value in predicting whether CD is active or not. |
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