摘要
Abstract
Objective To explore the possible targets for the development of the irritable bowel syndrome by screening and analysing the differentially expressed genes based on bioinformatics methods and verify them in microglial.Methods The gene expression microarray data of GSE36701 and GSE166869 were downloaded from GEO database and divided into irritable bowel syndrome(IBS)group and healthy control(HC)group.The data were processed using the relevant software package of R language,and the differentially expressed genes(DEGs)were obtained and enriched.Then the common differentially expressed genes were obtained from the intersec-tion.The g:Profiler database was used to analyze the common differentially expressed genes by GO and KEGG.The STRING database was used to analyze the protein interaction of the common differentially expressed genes.The key genes were screened by Cytoscape software.The WebGestalt database was used to analyze the enrichment of the up-regulated differentially expressed genes related to diseases and drugs.The microglial models of stress,inflammation and stress plus inflammation were constructed using cortisterone(Cort),lipopolysaccharide(LPS)and Cort plus LPS,respectively.GABRE and NF-κB were detected by Western blotting and quanti-tative real-time PCR,and the expressions of inflammatory factors were detected by ELISA.Results The results of microarray analysis using R language software package showed that there were 66 differentially expressed genes.GSEA analysis showed that IBS was associated with Parkinson's disease,Alzheimer's disease and other diseases.GO analysis showed that the differentially expressed genes were mainly enriched in biological processes such as natural killer cell-mediated cytotoxic regulation and natural killer cell-mediated immune regulation,in cell components such as tertiary granular lumen and microvillus,and in molecular functions such as lamin binding and aminoacyltransferase activity.KEGG analysis showed that the common differentially expressed genes were enriched in γ-GABAergic synapses and other pathways.Protein interaction analysis showed that CDC20,PTTG1 and BIRC5 were the key genes.The prediction analysis of up-regulated differentially expressed genes and disease showed that IBS was associated with hyperappetence,schizoaffective disorder and other diseases.The prediction analysis of up-regulated differentially expressed genes and drug showed that IBS was related to flunitrazepam,anti-anxiety drugs and other drugs.Western blotting and quantitative real-time PCR showed that the expressions of GABRE,TLR4,IκBα,NF-κB p65,TNF-α,IL-1β in LPS group,Cort group and LPS+Cort group were significantly higher than those in sham group,and the expressions in LPS+Cort group were significantly higher than those in LPS group and Cort group(P＜0.05).ELISA results showed that the expressions of TNF-α,IL-6 and CRP in LPS group,Cort group and LPS+Cort group were significantly higher than those in sham group(P＜0.001),and the expressions in LPS+Cort group were significantly higher than those in LPS group and Cort group(P＜0.001).Conclusion GABRE and MICALL2 genes are associated with the development of IBS and are potential targets for IBS treatment.关键词
肠易激综合征/差异表达基因/富集分析/小胶质细胞/应激/炎症Key words
irritable bowel syndrome/differential expression genes/enrichment analysis/microglia/stress/inflammation分类
医药卫生
