非洲猪瘟病毒感染后猪外周血淋巴细胞Toll样受体信号通路的lncRNA-mRNA分析

【目的】丰富非洲猪瘟病毒（African swine fever virus，ASFV）感染后猪外周血淋巴细胞长链非编码RNA（long non-coding RNA，lncRNA）表达谱，并进一步挖掘影响Toll样受体信号通路的调控网络。【方法】试验动物感染ASFV，于第7天采集外周血并分离得到外周血淋巴细胞，运用Illumina高通量组学测序对外周血淋巴细胞中lncRNA进行测序，原始数据经处理后筛选获得差异表达的lncRNA，并进行靶基因预测，利用生物信息学方法对靶基因进行GO功能和KEGG信号通路富集分析，初步绘制与Toll样受体信号通路相关的lncRNA-mRNA调控网络，并对lncRNA-ENSSSCG00000041959在内的4个lncRNAs进行实时荧光定量RT-PCR验证。【结果】共筛选到73个差异表达的lncRNAs，其中上调表达lncRNAs 38个，下调表达lncRNAs 35个。GO功能分析结果显示，靶基因显著富集在调节免疫系统过程、防御反应、生物刺激、病毒反应和先天性免疫；KEGG信号通路富集分析显示，大部分靶基因与细胞循环、疾病及免疫应答有关，其中免疫相关信号通路有Toll样受体信号通路、TNF信号通路、产生IgA的肠道免疫网络等。进一步挖掘出lncRNA-ENSSSCG00000041959-RIPK1和lncRNA-ENSSSCG00000041959-IRAK1可能是影响Toll样受体信号通路的重要调控网络，实时荧光定量RT-PCR与测序结果一致。【结论】本研究初步鉴定出lncRNA-ENSSSCG00000041959-RIPK1和lncRNA-ENSSSCG00000041959-IRAK1可能是影响Toll样受体信号通路的lncRNA-mRNA调控网络，为进一步探索lncRNA调控ASFV感染机体免疫反应奠定了理论基础。

lncRNA-mRNA Analysis of Toll-like Receptor Signaling in Porcine Peripheral Blood Lymphocytes After African Swine Fever Virus Infection

【Objective】 This study was aimed to enrich the long non-coding RNA (lncRNA) expression profile of pig peripheral blood lymphocytes after African swine fever virus (ASFV) infection,and further explore the regulatory network affecting the Toll-like receptor signaling pathway.【Method】 Experimental animals were infected with ASFV, and peripheral blood was collected on the 7th day and isolated to obtain peripheral blood lymphocytes.The lncRNAs in peripheral blood lymphocytes were sequenced by Illumina high-throughput genomic sequencing.The raw data were processed and screened to obtain differentially expressed lncRNA,and target gene prediction was performed.GO function and KEGG signaling pathway enrichment analysis were performed using bioinformatics methods,and preliminary mapping of the lncRNA-mRNA regulatory networks related to the Toll-like receptor signaling pathway and 4 lncRNAs including lncRNA-ENSSSCG00000041959 were verified by Real-time quantitative RT-PCR.【Result】 A total of 73 differentially expressed lncRNAs were screened,including 38 upregulated and 35 downregulated lncRNAs.GO function analysis showed that target genes were significantly enriched in regulating immune system process,defense response,biological response,viral response and innate immunity.KEGG signaling enrichment analysis showed that most target genes were related to cell circulation,disease and immune response,including immune related signaling were Toll-like receptor signaling pathway,TNF signaling pathway,and intestinal immune network producing IgA.lncRNA-ENSSSCG00000041959-RIPK1 and lncRNA-ENSSSCG00000041959-IRAK1 might be important regulatory networks affecting Toll-like receptor signaling pathway.The result of Real-time quantitative RT-PCR was consistent with those of sequencing.【Conclusion】In this study,lncRNA-ENSSSCG00000041959-RIPK1 and lncRNA-ENSSSCG00000041959-IRAK1 were initially identified as the lncRNA-mRNA regulatory network affecting the Toll-like receptor signaling pathway,which laid a theoretical foundation for further exploration of lncRNA regulation of the immune response of ASFV infection.