To explore immune-related mechanisms and potential gene targets of brucellosis spondylitis(BS)based on bioinformatics
Objective To search for differentially expressed genes(DEGs)and key signaling pathways in intervertebral tissues of the patients with brucellosis spondylitis(BS)through bioinformation analysis,so as to provide theoretical basis for in-depth understanding of immune-related mechanisms of BS and search for new therapeutic targets.Methods Intervertebral tissues of 4 patients with BS and 3 patients with lum-bar disc herniation(LDH)were collected for transcriptome sequencing.Bioinformatics analysis was used to find DEGs in the interbody tissues of BS patients and LDH patients.The Enirchr online enrichment tool was used to conduct gene ontological(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis for the screened differentially expressed im-mune genes.Protein-protein interaction(PPI)networks with Cytoscape software and screen was construc-ted to identify key genes.Intervertebral tissues of 14 patients with BS(BS group)and 10 patients with LDH disease(control group)were collected for qRT-PCR to verify the biological information analysis re-sults.Meanwhile,peripheral blood of the patients was collected,and the proportion of M2 macrophages and Th2 cells in peripheral blood of the patients in the 2 groups was detected by flow cytometry.Results The proportion of M2-type macrophages and Th2 cells in the intervertebral tissue of BS patients was higher than that of LDH patients(P＜0.05).A total of 1 651 DEGs were screened,with 1 294 up-regulated genes and 357 down-regulated genes.The biological processes of 1 651 DEGs were mainly concentrated in antigen processing and presentation,immune response,immune system process,etc.The cellular compo-nents were mainly concentrated in MHC protein complex,MHC Class Ⅱ protein complex and important components of cell membrane and the molecular functions were mainly related to phosphatase activity,polysaccharide binding and ion channel activity.The enrichment analysis of KEGG pathway showed that the PI3K-Akt signaling pathway,calcium signaling pathway and MAPK signaling pathway were closely re-lated to the development of BS.Through PPI network analysis,the top 20 key genes were selected and in-tersected with the genes of the PI3K-Akt signaling pathway to obtain 3 key genes,namely FN1,EGFR and EGF.qRT-PCR results showed that the mRNA expressions of FN1,EGFR and EGF were higher than those of control group(P＜0.000 1).Conclusion FN1,EGFR and EGF may be biomarkers and potential therapeutic targets for BS,and this study is helpful to strengthen the understanding of the immunomodu-latory mechanism related to BS.
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