Th2倾向型典型哮喘与咳嗽变异性哮喘外周血单个核细胞基因表达谱的差异性研究  

Differential gene expressions in peripheral blood mononuclear cells between Th2-driven classical asthma and cough variant asthma

作　　者：杜敏 魏维 周光红 黄静[1] 王玉霞[2] 龙怀聪[2] DU Min;WEI Wei;ZHOU Guanghong;HUANG Jing;WANG Yuxia;LONG Huaicong(North Sichuan Medical College,Nanchong,Sichuan 637000,P.R.China;Geriatric Intensive Care Unit,Sichuan Academy of Medical Sciences&Sichuan Provincial People's Hospital,School of Medicine,Chengdu,Sichuan 610072,P.R.China;Department of Respiratory and Critical Care Medicine,Anyue County People's Hospital,Anyue,Sichuan 642350,P.R.China;Department of Respiratory and Critical Care Medicine,Sichuan Academy of Medical Sciences&Sichuan Provincial People's Hospital,School of Medicine,Chengdu,Sichuan 610072,P.R.China)

机构地区：[1]川北医学院,四川南充637000 [2]四川省医学科学院·四川省人民医院老年重症监护室,四川成都610072 [3]安岳县人民医院呼吸与重症医学科,四川安岳642350 [4]四川省医学科学院·四川省人民医院呼吸与危重症学科,四川成都610072

出　　处：《中国呼吸与危重监护杂志》2023年第6期395-402,共8页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：四川省学术和技术带头人培养支持资助项目[川人社办发(2017)919号]。

摘　　要：目的揭示Th2倾向型典型哮喘(classical asthma,CA)与咳嗽变异性哮喘(cough variant asthma,CVA)在基因表达水平上的差异,以便进一步研究哮喘的发病机制。方法收集2018年6月1日—2019年12月31日在四川省人民医院呼吸与危重症学科门诊就诊的哮喘患者,健康对照(healthy control,HC)为体检中心的健康成人。收集临床数据,用全长转录组测序方法检测CA组、CVA组和HC组中外周血单个核细胞基因表达情况。通过GO、KEGG分析和蛋白-蛋白相互作用(protein-protein interaction,PPI)网络分析筛选差异基因。通过Cytoscape软件对互作网络分析结果进行可视化处理。最后通过实时荧光定量聚合酶链反应(real-time polymerase chain reaction,RT-PCR)对候选基因进行验证。结果研究共纳入31例哮喘患者,其中CA组20例,CVA组11例。根据血清总IgE>60 IU/mL和呼出气一氧化氮分数>40 ppb为筛选条件,筛选出Th2倾向型CA 9例、Th2倾向型CVA 5例进行分析。基因表达分析显示,在Th2倾向型CA组和Th2倾向型CVA组之间共有差异基因300个,其中155个基因相对上调,145个基因相对下调。GO富集分析表明,差异基因主要富集在药物反应、氮化合物生物合成、细胞质基质、蛋白质结合、ATP结合等方面。KEGG通路分析显示,差异基因主要富集在2-氧羧酸代谢和自然杀伤细胞介导的细胞毒信号通路中。PPI分析显示不同基因之间存在广泛的蛋白互作。筛选了10个候选基因并进行RT-PCR验证,最终发现CLU、GZMB、PPBP、PRF1、PTGS1、TMSB4X在Th2倾向型CA组和Th2倾向型CVA组间存在显著性差异表达。结论哮喘的发生是众多基因和通路相互作用的结果,CLU、GZMB、PPBP、PRF1、PTGS1、TMSB4X基因可能在Th2倾向型CVA向Th2倾向型CA的发展过程中起着重要作用。Objective To reveal the differences in gene expression levels between Th2-driven classical asthma(CA)and Th2-driven cough variant asthma(CVA)in order to investigate the pathogenesis of asthma further.Methods Clinical data were collected from asthmatic patients in the Department of Respiratory and Critical Care of Sichuan Provincial People's Hospital from June 1,2018,to December 31,2019.The healthy control(HC)group was healthy adults from the physical examination center.Gene expression of peripheral blood mononuclear cells(PBMCs)in the CA group,CVA group,and HC group was determined by full-length transcriptome sequencing.Differential genes were screened by GO,KEGG analysis,and protein-protein interaction(PPI)network analysis.The results of interaction network analysis were visualized by Cytoscape.Finally,the candidate genes were verified by real-time quantitative polymerase chain reaction(RT-PCR).Results A total of 31 patients with asthma were included in the study,including 20 patients in the CA group and 11 patients in the CVA group.According to serum total IgE>60 IU/mL and fractional exhaled nitric oxide(FeNO)>40 ppb as the screening condition,9 cases of Th2-driven CA and 5 cases of Th2-driven CVA were screened for analysis.Gene expression analysis showed 300 differentially expressed genes between the Th2-driven CA group and the Th2-driven CVA group,among which 155 genes were up-regulated,and 145 were downregulated.GO enrichment analysis showed that differential genes were mainly enriched in drug response,nitrogen compound biosynthesis,cytoplasmic matrix,protein binding,ATP binding,etc.KEGG pathway analysis showed that differential genes were mainly concentrated in 2-oxy-carboxylic acid metabolism and cytotoxic signaling pathways mediated by natural killer cells.PPI analysis revealed extensive protein interactions between different genes.Ten candidate genes were screened for RT-PCR verification and finally found that CLU,GZMB,PPBP,PRF1,PTGS1,and TMSB4X were significantly differentially expressed between the Th2

关 键 词：典型哮喘 咳嗽变异性哮喘 差异表达基因 

分 类 号：R562.25[医药卫生—呼吸系统]