Expression profile of microRNAs in porcine alveolar macrophages after Toxoplasma gondii infection
Expression profile of microRNAs in porcine alveolar macrophages after Toxoplasma gondii infection
复制标题
弓形虫感染后猪肺泡巨噬细胞中 microRNA 的表达谱
DOI：
10.1186/s13071-019-3297-y
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发表时间：
2019-01-29
影响因子：
3.2
通讯作者：
Fang, Rui
中科院分区：
文献类型：
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作者：
Li, Senyang;Yang, Jing;Fang, Rui
刚地弓形虫是一种顶复门原虫寄生虫，可引起人类和动物严重的临床疾病。microRNA（miRNAs）是一类非蛋白质编码的RNA，可调控靶基因的表达。先前的研究发现，许多miRNAs在T.弓形虫感染和发挥显着的影响，并揭示了宿主的生存和不同菌株的毒力可以通过不同的miRNA调节。在T.弓形虫感染，但很少有研究探讨miRNA与猪肺泡巨噬细胞感染弓形虫的关系。刚地。用RH（Ⅰ型）和Me49（Ⅱ型）菌株感染猪肺泡巨噬细胞（3D4 - 21）。培养12 h和24 h后收获。使用用于Illumina®的NEBNext® Multiplex Small RNA Library Prep Set（NEB，USA）生成miRNA文库，并且基于每百万读段的转录本（TPM）估计miRNA表达水平。我们的研究产生了6个miRNA表达谱与RH和Me49感染的巨噬细胞与对照组相比。比较了T.感染和未感染弓形虫的样本鉴定出81个差异表达的miRNAs，包括36个新的miRNAs和45个成熟的miRNAs。利用miRanda软件对差异表达的miRNAs的靶基因进行预测，发现ssc-miR-127和ssc-miR-143 - 3p分别调控一氧化氮合酶1（NOS1）和一氧化氮合酶3（NOS3），这两种酶在氧化L-精氨酸合成一氧化氮（NO）过程中起重要作用。这些基因在RH和Me49感染组中差异表达。KEGG富集分析表明，所预测的靶基因参与多种信号通路，包括Fc γ R介导的吞噬作用、AMPK信号通路、mTOR信号通路和Fc γ RI信号通路，所有这些都是猪肺泡巨噬细胞正常功能不可或缺的。我们的研究结果提供了猪肺泡巨噬细胞感染T.刚地。据我们所知，这项研究首次证明了miRNA与猪源性巨噬细胞之间的关系。了解这些受调控的miRNAs的功能将有助于T.差异表达的miRNAs可能是弓形虫感染性疾病的候选药物靶点。猪的弓形虫感染
Toxoplasma gondii is an apicomplexan protozoan parasite that can cause serious clinical illnesses in both humans and animals. microRNAs (miRNAs) are non-protein-coding RNAs that can regulate the expression of target genes. A previous study found that many miRNAs were differentially expressed after T. gondii infection and exert significant effects and revealed that both host survival and the virulence of different strains can be regulated by different miRNAs. Macrophages play an important role in T. gondii infection, but few studies have investigated the relationship between miRNAs and porcine alveolar macrophages infected with T. gondii. Porcine alveolar macrophages (3D4-21) were infected with the RH (Type I) and Me49 (Type II) strains of T. gondii for 12 h and 24 h and then harvested. miRNA libraries were generated using the NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (NEB, USA), and the miRNA expression levels were estimated based on transcripts per million reads (TPM). Our study generated six miRNA expression profiles from macrophages infected with RH and Me49 compared with the control groups. The comparison of the T. gondii-infected and uninfected samples identified 81 differentially expressed miRNAs, including 36 novel miRNAs and 45 mature miRNAs. The target genes of these differentially expressed miRNAs were predicted using miRanda software, and ssc-miR-127 and ssc-miR-143-3p were predicted to regulate nitric oxide synthase 1 (NOS1) and nitric oxide synthase 3 (NOS3), respectively, which play essential roles in synthesizing nitric oxide (NO) by oxidizing L-arginine. These genes were differentially expressed in both the RH- and Me49-infected groups. A KEGG enrichment analysis indicated that the predicted target genes were involved in multiple signaling pathways, including FcγR-mediated phagocytosis, the AMPK signaling pathway, the mTOR signaling pathway, and the FcγRI signaling pathway, all of which are indispensable for the normal functioning of porcine alveolar macrophages. Our results provide data on the miRNA profile of porcine alveolar macrophages infected with T. gondii. To our knowledge, this study provides the first demonstration of the relationship between miRNA and macrophages of swine origin. Understanding the functions of these regulated miRNAs will aid the investigation of T. gondii infectious diseases, and the differentially expressed miRNAs might be candidate drug targets for T. gondii infection in pigs.