45例麻风病患者外周血CD4^(+) T细胞差异表达基因筛查  

Screening of differentially expressed genes in CD4^(+)T cells in peripheral blood of 45 leprosy patients

作　　者：陈欢[1] 尹江柳 Chen Huan;Yin Jiangliu(Department of Leprosy Prevention and Control,Hunan Provincial Center for Disease Control and Prevention,Changsha 410005,China;Department of Neurosurgery,Changsha Central Hospital,University of South China,Changsha 410006,China)

机构地区：[1]湖南省疾病预防控制中心麻风病防治科,长沙410005 [2]南华大学附属长沙中心医院神经外科,410006

出　　处：《中华皮肤科杂志》2021年第8期683-687,共5页Chinese Journal of Dermatology

基　　金：湖南省科技创新计划项目(2018JJ6010)。

摘　　要：目的检测麻风病患者外周血CD4^(+)T细胞中mRNA表达谱,筛选并验证可能与麻风病发病过程密切相关的基因。方法2018年7月至2020年5月在湖南省收集45例麻风病患者及45例健康人,以磁珠法分离外周血CD4^(+)T细胞,提取RNA。在上述受试者中,随机选取6例患者及6例健康人,采用Solexa测序法筛查两组间差异表达基因,将差异倍数2倍以上者(P<0.05)定义为差异表达基因,并进行KEGG Pathway富集分析,再以实时荧光定量PCR验证基因的表达水平。结果基因筛查发现4831个转录本,属于新基因且具有蛋白编码潜能,在两组间筛选出8个差异表达基因,其中,CXCL8、PPBP、RPS18及IL-1β基因mRNA表达水平上调,RNH1、RPL39、RPL15及AMBRA1基因mRNA表达水平下调。实时荧光定量PCR验证结果与上述基因筛查结果一致。KEGG分析显示,麻风病患者与健康对照组差异表达基因主要富集在线粒体自噬及细胞自噬相关通路和人乳头状瘤病毒感染通路。结论麻风病患者AMBRA1、RNH1基因mRNA表达水平下调,CXCL8、PPBP、IL-1β基因mRNA表达水平上调,可能分别通过线粒体自噬通路和趋化因子介导的信号通路参与麻风病发病。Objective To detect the mRNA expression profile of CD4^(+)T cells in the peripheral blood of leprosy patients,and to screen and identify genes that may be closely related to the pathogenesis of leprosy.Methods From July 2018 to May 2020,45 leprosy patients were collected from Hunan Province,and 45 healthy volunteers from Health Examination Center of Changsha Central Hospital.CD4^(+)T cells were isolated from peripheral blood samples by using magnetic beads,and then RNA was extracted.Solexa sequencing was performed to screen differentially expressed genes between 6 patients and 6 healthy controls,who were randomly selected from the above subjects.Differentially expressed genes were defined as those with a fold change greater than 2 and a P value below 0.05,and then Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis was performed.Real-time fluorescence-based quantitative PCR was conducted to verify the gene expression.Results Genetic screening revealed 4831 newly-discovered transcripts with protein-coding potential.Eight differentially expressed genes were screened out between the two groups.Among them,the mRNA expression of CXCL8,PPBP,RPS18 and IL-1βgenes was up-regulated,while the mRNA expression of RNH1,RPL39,RPL15 and AMBRA1 genes was down-regulated.Verification results of real-time fluorescence-based quantitative PCR were consistent with the above-mentioned genetic screening results.KEGG analysis showed that the differentially expressed genes between leprosy patients and healthy controls were mainly enriched in mitochondrial autophagy,autophagy-related pathways and human papillomavirus infection pathways.Conclusion Down-regulated mRNA expression of AMBRA1 and RNH1 genes and up-regulated mRNA expression of CXCL8,PPBP and IL-1β genes were identified in patients with leprosy,which may be involved in the pathogenesis of leprosy through the mitochondrial autophagy pathway and chemokine-mediated signaling pathway,respectively.

关 键 词：麻风 CD4阳性T淋巴细胞 基因表达谱 自噬 α乳头状瘤病毒属 

分 类 号：R755[医药卫生—皮肤病学与性病学]