摘要
目的基于GEO数据库数据分析原发性干燥综合征(pSS)外周血和唇腺组织中差异表达基因的生物学功能,筛选调控pSS发生发展的关键基因,并观察pSS患者唇腺组织中的免疫细胞变化情况。方法从GEO数据库获取pSS相关基因表达谱芯片GSE51092,利用R软件筛选出pSS患者外周血和唇腺组织中的差异表达基因,并进行GO功能富集分析、KEGG信号通路富集分析、基因集富集分析(GSEA),观察差异表达基因的生物学功能。通过STRING数据库构建差异表达基因的蛋白互作网络,利用Cytoscape软件筛选连接度值排序前10的差异表达基因。通过CIBERSORT反卷积算法获取22种免疫细胞的细胞占比,分析各类免疫细胞间的相关性,并观察pSS患者和健康人之间的免疫细胞浸润差异。结果筛选出282个差异表达基因。GO功能富集分析结果显示,差异表达基因在生物学过程中体现在病毒应答、病毒防御应答、Ⅰ型干扰素(IFN)信号通路传导等过程上;分子功能变化主要集中在双链RNA结合、翻译调节活性、NAD+ADP−核糖转移酶活性等功能上。KEGG信号通路富集分析发现,显著富集的信号通路包括冠状病毒(COVID)-19、人乳头瘤病毒感染、甲型流感等。GSEA分析结果显示,差异表达基因富集在免疫反应通路,包括对病毒的防御反应、对IFN-γ的应答、RIG-I样受体信号通路、调控破骨细胞分化等通路。连接度值排序前10的基因为STAT1、IRF7、CXCL10、MX1、ISG15、IFIT3、IFIH1、OAS1、OAS2、IFIT2。pSS患者唇腺组织中细胞占比较高的免疫细胞类型分别是M2巨噬细胞、M0巨噬细胞、M1巨噬细胞、幼稚B细胞、活化的CD4+T细胞、活化的NK细胞,其中活化的NK细胞与CD8+T细胞的细胞占比呈正相关关系(r=0.44,P<0.05),活化的记忆CD4+T细胞与Treg的细胞占比呈负相关关系(r=-0.54,P<0.05)。与健康人相比,pSS患者唇腺组织中活化树突状细胞(DC)的细胞占比显著增加(P<0.001),CD8+T细胞的细胞占比显著降低(P=0.038)。结论与健康人相比,pSS患者外周血和唇腺组织中有282个差异表达基因;差异表达基因与病毒感染、I型IFN信号通路传导、对病毒的防御反应等免疫反应通路有关,STAT1、IRF7、CXCL10等10个基因可能是pSS发生发展的关键调控基因。pSS患者唇腺组织中活化的NK细胞与CD8+T细胞的细胞占比正相关,活化的记忆CD4+T细胞与Treg的细胞占比负相关;与健康人相比,pSS患者唇腺组织中活化DC的细胞占比增加、CD8+T细胞的细胞占比降低。
Objective The biological functions of differentially expressed genes in the peripheral blood and labial glandular tissues of patients with primary Sjogren's syndrome(pSS)were analyzed based on GEO database,the key genes that regulated the occurrence and development of pSS were screened,and the changes of immune cells in the labial glandular tissues of pSS patients were observed.Methods The expression profile chip GSE51092 of pSS-related genes was obtained from GEO database.The differentially expressed genes in the peripheral blood and labial gland tissues were screened out by R software,and the biological functions of the differentially expressed genes were observed by GO function enrichment analysis,KEGG signal pathway enrichment analysis and gene set enrichment analysis(GSEA).The protein interaction network of differentially expressed genes was constructed through STRING database,and the top 10 differentially expressed genes were screened out by Cytoscape software.The cell proportion of 22 kinds of immune cells was obtained by CIBERPORT deconvolution algorithm,the correlation between various immune cells was analyzed,and the difference of immune cell infiltration between pSS patients and healthy people was observed.Results A total of 282 differentially expressed genes were screened out.The results of GO function enrichment analysis showed that the differentially expressed genes were reflected in the processes of response to virus,defense response to virus,type I interferon(IFN)signal pathway transmission,etc.The changes of molecular functions mainly focused on double stranded RNA binding,translation regulation activity,NAD+ADP-ribotransferase activity and other functions.KEGG signal pathway enrichment analysis found that the significantly enriched signal pathways included COVID-19,human papillomavirus infection,influenza A,etc.GSEA analysis showed that the gene set was concentrated in pathways such as defense response to the virus,response to IFN-γ,RIG-I-like receptor signaling pathway,regulation of osteoclast differentiation,etc.The top 10 genes with connectivity values were STAT1,IRF7,CXCL10,MX1,ISG15,IFIT3,IFIH1,OAS1,OAS2,and IFIT2.The immune cell types with high cell proportion in the labial gland tissues of pSS patients were M2 macrophages,M0 macrophages,M1 macrophages,immature B cells,activated CD4+T cells,and activated NK cells,in which,the activated NK cells were positively correlated with the cell proportion of CD8+T cells(r=0.44,P<0.05),and the activated memory CD4+T cells were negatively correlated with the cell proportion of Treg(r=-0.54,P<0.05).Compared with the healthy people,the proportion of activated dendritic cells(DC)in the labial gland tissues of pSS patients significantly increased(P<0.001),and the proportion of CD8+T cells significantly decreased(P=0.038).Conclusions Compared with healthy people,there are 282 differentially expressed genes in the peripheral blood and labial gland tissues of pSS patients.The differentially expressed genes are related to immune response pathways such as virus infection,type I IFN signal pathway transmission,and defense response to virus.Ten genes such as STAT1,IRF7 and CXCL10 may be the key regulatory genes for the development and progression of pSS.The proportion of activated NK cells and CD8+T cells in the labial gland of pSS patients are positively correlated,and the proportion of activated memory CD4+T cells and Treg cells are negatively correlated.Compared with healthy people,the proportion of activated DC cells in labial gland tissues of pSS patients increases,and the proportion of CD8+T cells decreases.
作者
饶钰君
田龙龙
张勇斌
闵星星
陈桂玉
李国青
RAO Yujun;TIAN Longlong;ZHANG Yongbin;MIN Xingxing;CHEN Guiyu;LI Guoqing(Department of Rheumatology and Immunology,The Affiliated Hospital of Yangzhou University,Yangzhou 225000,China;不详)
出处
《山东医药》
CAS
2022年第21期48-53,共6页
Shandong Medical Journal
基金
国家自然科学基金面上项目(82071821)
江苏省自然科学基金面上项目(BK20201222)。
关键词
原发性干燥综合征
差异表达基因
生物学功能
免疫细胞浸润
primary Sjogren's syndrome
differentially expressed genes
biological function
immune cell infiltration
作者简介
通信作者:李国青(E-mail:Guoqing1007@163.com)。
