1.The First Clinical College, Hubei University of Chinese Medicine, Hubei Province, Wuhan 430061, China;
2.Department of Rheumatism Immunology, Hubei Provincial Hospital of Traditional Chinese Medicine, Hubei Province, Wuhan 430061, China;
3.Xiangnan Famous Medical Studio, Hubei Province, Wuhan 430061, China;
4.Department of Geriatrics, Hubei Provincial Hospital of Traditional Chinese Medicine, Hubei Province, Wuhan 430061, China;
5.Department of Endocrine, Hubei Provincial Hospital of Traditional Chinese Medicine, Hubei Province, Wuhan 430061, China;
6.College of Basic Medical, Hubei University of Chinese Medicine, Hubei Province, Wuhan 430061, China;
7.College of Acupuncture and Orthopedics, Hubei University of Chinese Medicine, Hubei Province, Wuhan 430061, China
Abstract:Objective GEO database was used to conduct differential analysis, enrichment analysis, and immune infiltration analysis of gene data of thyroid-associated ophthalmopathy, providing theoretical basis for in-depth understanding of the immune related mechanism of thyroid-associated ophthalmopathy. Methods “Graves disease” was retrieved from GEO database, and orbital expression genes before thyroid-associated ophthalmopathy were selected as experimental group, and normal anterior orbital expression genes were used as control group. PPI analysis of differential genes was performed using STRING database to screen core genes; the R software was used for GO and KEGG enrichment analysis of differential genes; meanwhile, CIBERSORT deconvolution method was used to analyze the infiltration of 22 kinds of immune cells in thyroid-associpated ophthalmopathy. Results According to GEO database, 315 differential genes were obtained, including 220 down-regulated genes and 95 up-regulated genes, 56 core genes were screened by protein-protein interaction (PPI) analysis, and the top five core genes were FOS, JUN, CXCL8, ITGAM, and CXCL12, respectively; GO enrichment analysis showed 460 biological processes, six cellular components, and 36 molecular functions, it mainly involved cell chemotaxis, leukocyte chemotaxis, granulocyte chemotaxis, leukocyte migration, monocyte migration, and other biological processes; KEGG enrichment analysis showed 20 signaling pathways, mainly involved cytokine-cytokine receptor interaction, PI3K-Akt signaling pathway, interleukin-17 signaling pathway, NF-κB signaling pathway, Toll-like receptor signaling pathway, osteoclast differentiation, and other signaling pathways; the results of immune infiltration showed that monocytes, plasma cells and neutrophils were significantly increased in the experimental group, while M2 macrophages and inactive resting mast cells were significantly decreased. Immunocell correlation analysis showed a significant positive correlation between plasma cells and activated mast cells (r = 0.64), and there was a significant positive correlation between monocytes, inactive natural killer cells and neutrophils (r = 0.61, 0.62), regulatory T cells were significant positively correlated with CD8+T cells (r = 0.60), there was a significant negative correlation between inactive resting mast cells, plasma cells and activated mast cells (r = -0.71, -0.68), there was a significant negative correlation between monocytes, M2 macrophages and inactive resting mast cells (r = -0.71, -0.66), there was a significant negative correlation between inactive and activated natural killer cells (r = -0.71), the difference analysis of immune infiltration between experimental group and control group indicated that monocytes and neutrophils in experimental group were significantly increased (P < 0.05), M2 macrophages and non-activated resting mast cells were significantly decreased (P < 0.05). Conclusion There are various kinds of immune cell infiltration in thyroid-associated ophthalmopathy, immune response and inflammatory reaction throughout the disease.
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