基于生物信息学分析原发性干燥综合征的关键基因和免疫浸润机制及靶向中药预测  

Analysis of Key Genes and Immune Infiltration Mechanism of Primary Sjögren's Syndrome and Prediction of Targeted Traditional Chinese Medicine Based on Bioinformatics

作　　者：袁心柱 李玲琴[2] 杜欢 林昌伟[1] 袁祖君 王彦江[1] 王宝福[1] 谢席胜[1] Yuan Xinzhu;Li Lingqin;Du Huan;Lin Changwei;Yuan Zujun;Wang Yanjiang;Wang Baofu;Xie Xisheng(The Affiliated Nanchong Central Hospital of North Sichuan Medical College,Nanchong 637000,China;The Affiliated of North Sichuan Medical College,Nanchong 637000,China;Traditional Chinese Medicine Hospital of Xichong County,Nanchong 637200,China)

机构地区：[1]川北医学院附属南充市中心医院,南充637000 [2]川北医学院附属医院,南充637000 [3]西充县中医医院,南充637200

出　　处：《世界科学技术-中医药现代化》2023年第11期3592-3604,共13页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：四川省中医药管理局科学技术研究专项课题(2020JC0079):黄芪三七合剂通过PINK1/Parkin通路调控线粒体自噬减轻慢性肾脏病小鼠炎症及肾脏纤维化,负责人:谢席胜

摘　　要：目的利用生物信息学方法探讨原发性干燥综合征(Primary Sj?gren’s syndrome,pSS)的关键基因,分析其可能的免疫浸润机制,并挖掘潜在的靶向中药,为pSS的临床治疗提供新方向。方法从GEO数据库下载pSS基因表达谱芯片数据集,使用R软件进行差异基因的筛选,并对这些差异基因进行基因本体论(GO)及基因通路富集(KEGG)富集分析。应用STRING数据库对差异基因进行蛋白质互作网络分析,利用Cytoscape筛选核心基因,应用ELISA方法对关键基因表达进行验证。通过CIBERSORT反卷积法计算22种免疫细胞在pSS中的相对表达情况。最后通过Coremine Medical数据库对靶向pSS关键基因的中药进行预测。结果筛选出pSS差异基因共计232个,其中上调207个,下调25个。GO主要富集于:白细胞介导的免疫、淋巴细胞介导的免疫、白细胞-细胞粘附等;KEGG通路主要富集于:造血细胞谱系、原发免疫缺陷、产生IgA的肠道免疫网络、吞噬体、利什曼病等。筛选出:PTPRC、CD19、LCP2、CCR5、CD69等10余个关键基因,实验验证核心基因表达与芯片数据的表达趋势一致。免疫浸润结果显示:记忆B细胞、活化的CD4+记忆T细胞、初始CD4 T细胞在pSS中表达显著上调。免疫细胞相关性分析显示:单核细胞和调节性T细胞呈中度正相关(r=0.63),M1型巨噬细胞和初始B细胞呈中度正相关(r=0.52),而浆细胞和活化的CD4+记忆性T细胞呈高度负相关(r=0.75)。通过COREMINE Medical预测发现,人参、三七、雷公藤、地榆、厚朴、马钱子等可能靶向治疗pSS。结论pSS的发生与发展是多基因多通路共同参与的结果,记忆B细胞、活化的CD4+记忆T细胞、初始CD4 T细胞可能促进了pSS的发展。人参、三七、雷公藤、地榆、厚朴、马钱子等可能为潜在治疗pSS的靶向中药。Objective Using bioinformatics methods to study the immune infiltration mechanism of Primary Sjögren's syndrome(pSS)and to explore potential target Chinese medicines,which can provide new directions for the clinical treatment of pSS.Methods Gene expression profile microarray dataset of pSS was downloaded from the GEO database,differential genes were screened using R software,and gene ontology(GO)and gene pathway enrichment(KEGG)enrichment analysis was performed on these differential genes.Protein interaction network analysis of differential genes was performed by applying the STRING database,key genes were screened by using Cytoscape,and ELISA for the verification of key genes expression.Immune infiltration and correlation of immune cells in pSS were calculated by CIBERSORT inverse convolution method in 22.Finally,the herbal prediction of key target genes was performed by using the Coremine Medical database.Results A total of 232 differential genes were obtained,of which 207 were upregulated and 25 were down-regulated.GO was mainly enriched in:leukocyte mediated immunity,lymphocyte mediated immunity,leukocyte cell−cell adhesion,etc;KEGG was mainly enriched in Hematopoietic cell lineage,Primary immunodeficiency,Intestinal immune network for IgA production,Phagosome,Leishmaniasis.Ten key genes were screened:PTPRC,CD19,LCP2,CCR5 and CD69 etc.The hub genes expression in the pSS is the same as that of GSE40611.Immune infiltration showed that memory B cells,T cells CD4 memory activated,and T cells CD4 naïve were highly expressed in the pSS.Immune cell correlation analysis showed a positive correlation between initial Monocytes and T cells regulatory(Tregs),a positive correlation between Macrophages M1 and B cell naïve,and a negative correlation between Plasma cells and T cells CD4 memory activated.COREMINE Medical predicted that Ginseng,Panax notoginseng,Tripterygium wilfordii,Burnet,Magnolia,and Strychni may treat pSS.Conclusion The development and progression of pSS are the results of the combined involvement of

关 键 词：干燥综合征 GEO数据库 生物信息学 免疫浸润 

分 类 号：R-058[医药卫生]