日本血吸虫感染小鼠慢性致病阶段肝脏中差异表达长非编码RNA筛选及功能分析  

Screening and functional analysis of differentially expressed long non-cod-ing RNA in the liver of mice infected with Schistosoma japonicum during the chronic pathogenic stage

作　　者：李银龙 李琴 林伟娜 冯婷 秦志强 曹淳力 李石柱 许静 LI Yinlong;LI Qin;LIN Weina;FENG Ting;QIN Zhiqiang;CAO Chunli;LI Shizhu;XU Jing(National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases,National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention(Chinese Center for Tropical Diseases Research),National Health Commission Key Laboratory of Parasite and Vector Biology,WHO Collaborating Centre for Tropical Diseases,National Center for International Research on Tropical Diseases,Ministry of Science and Technology,Shanghai 200025,China;School of Global Health,Chinese Center for Tropical Diseases Research and Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)

机构地区：[1]传染病溯源预警与智能决策全国重点实验室、中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心)、国家卫生健康委员会寄生虫病原与媒介生物学重点实验室、WHO热带病合作中心、科技部国家级热带病国际联合研究中心,上海200025 [2]上海交通大学医学院-国家热带病研究中心全球健康学院,上海200025

出　　处：《中国血吸虫病防治杂志》2024年第2期137-147,共11页Chinese Journal of Schistosomiasis Control

基　　金：国家重点研发计划(2021YFC2300800,2021YFC2300804)。

摘　　要：目的 筛选日本血吸虫感染小鼠慢性致病阶段肝脏中差异表达长非编码RNA(long non-coding RNA, lnc RNA)并进行功能分析,为探索lnc RNA在日本血吸虫感染所致肝脏病变中的作用提供参考。方法 20只6周龄C57BL/6小鼠随机分为2组,每组10只。实验组每只小鼠采用腹部贴片法感染(15±2)条日本血吸虫尾蚴建立日本血吸虫慢性感染小鼠模型,以蒸馏水作为对照组。两组小鼠均于感染70 d后剖杀,获取小鼠肝脏组织样本,进行RNA抽提及文库构建。通过Illumina Nova Seq 6000测序平台对文库进行测序,采用fastp软件进行数据清洗,使用HISAT2软件进行参考基因组比对及基因表达量(FPKM)计算。采用CNIC、CPC、Pfam、PLEK软件对潜在lnc RNA序列进行编码潜能预测,筛选出潜在lnc RNA。采用DESeq2软件进行基因差异表达分析,筛选出差异表达lnc RNA。通过基因本体论(Gene Ontology,GO)和京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,挖掘差异表达lnc RNA靶基因参与的生物学过程和代谢途径。结果 共筛选出333个潜在lnc RNA,67个鉴定为差异表达lnc RNA,其中49个表达上调、18个表达下调。差异表达lnc RNA预测靶基因共53个,GO富集分析显示这些靶基因主要富集在生物学过程和分子功能;其中Sema7a、Arrb1、Ccl21b等基因可能是细胞外调节蛋白激酶1(extracellular regulated protein kinase,ERK1)和ERK2级联正调控生物学过程的关键靶基因,可能参与调控胶原蛋白表达。KEGG富集分析显示,差异表达lnc RNA靶基因主要参与细胞因子-细胞因子受体相互作用、病毒蛋白与细胞因子和细胞因子受体的相互作用、趋化因子信号通路和核因子κB(nuclear factor kappa-B,NF-κB)通路等信号通路。结论 本研究鉴定了日本血吸虫感染小鼠慢性致病阶段肝脏中差异表达lnc RNA及其靶基因的功能富集,其中上调表达的lnc RNA可能通过调控Sema7a、Arrb1、CObjective To screen differentially expressed long non-coding RNAs(lncRNAs)in the liver of mice infected with Schistosoma japonicum during the chronic pathogenic stage and identify their functions,so as to provide insights into unraveling the role of lncRNAs in S.japonicum infection-induced liver disorders.Methods Twenty 6-week-old C57BL/6 mice were randomly divided into two groups,of 10 animals each group.Each mouse in the experimental group was infected with(15±2)S.japonicum cercariae via the abdomen for modeling chronic S.japonicum infection in mice,and distilled water served as controls.All mice were sacrificed 70 days post-infection,and mouse liver specimens were sampled for RNA extraction and library construction.All libraries were sequenced on the Illumina NovaSeq 6000 sequencing platform.Data cleaning was performed using the fastp software,and reference genome alignment and gene expression(FPKM)calculation were performed using the HISAT2 software.Potential lncRNA sequences were predicted using the software CNIC,CPC,Pfam,and PLEK,and potential lncRNAs were screened.Differentially expressed lncRNAs were screened with the DESeq2 software and subjected to Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses to identify biological processes and metabolic pathways involved in target genes of differentially expressed lncRNAs.Results A total of 333 potential lncRNAs were screened,and 67 were identified as differentially expressed lncRNAs,including 49 up-regulated and 18 down-regulated lncRNAs.A total of 53 target genes were predicted for differentially expressed lncRNAs.GO enrichment analysis showed that these target genes were mainly enriched in biological process and molecular function,among which Sema7a,Arrb1,and Ccl21b genes may be hub target genes for positive regulation of extracellular regulated protein kinase 1(ERK1)and ERK2 cascades and may participate in the regulation of collagen expression.KEGG enrichment analysis showed that the target genes of differentially expresse

关 键 词：日本血吸虫 长非编码RNA 肝纤维化 细胞外调节蛋白激酶 小鼠 

分 类 号：R383.24[医药卫生—医学寄生虫学]