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绵羊布鲁氏菌在 RAW264.7 巨噬细胞免疫系统中存活的转录组全景。

Transcriptome Landscape of Intracellular Brucella ovis Surviving in RAW264.7 Macrophage Immune System.

机构信息

Immunology Research Center, Medical Research Institute, Southwest University, Chongqing, 402460, People's Republic of China.

College of Animal Science, Southwest University, Chongqing, 402460, People's Republic of China.

出版信息

Inflammation. 2020 Oct;43(5):1649-1666. doi: 10.1007/s10753-020-01239-4.

Abstract

Brucella ovis infection results in genital damage and epididymitis in rams, placental inflammation and rare abortion in ewes, and neonatal mortality in lambs. However, the mechanism underlying B. ovis infection remains unclear. In the present study, we used prokaryotic transcriptome sequencing to identify the differentially expressed genes (DEGs) between wild-type B. ovis and intracellular B. ovis in RAW264.7 macrophages. Gene ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed, and quantitative reverse transcriptase PCR (qRT-PCR) was used to validate the top 10 upregulated and downregulated DEGs. The results showed that 212 genes were differentially expressed, including 68 upregulated and 144 downregulated genes, which were mainly enriched in 30 GO terms linked to biological process, cellular component, and molecular function. KEGG analysis showed that the DEGs were enriched in the hypoxia-inducible factor 1 (HIF-1) signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, beta-alanine metabolism, and quorum sensing pathway. BME_RS01160, BME_RS04270, BME_RS08185, BME_RS12880, BME_RS25875, predicted_RNA865, and predicted_RNA953 were confirmed with the transcriptome sequencing data. Hence, our findings not only reveal the intracellular parasitism of B. ovis in the macrophage immune system, but also help to understand the mechanism of chronic B. ovis infection.

摘要

绵羊感染布鲁氏菌可导致公绵羊生殖器官损伤和附睾炎,母绵羊胎盘炎症和罕见流产,羔羊死亡。然而,绵羊布鲁氏菌感染的机制尚不清楚。本研究采用原核转录组测序技术,鉴定了 RAW264.7 巨噬细胞中野生型绵羊布鲁氏菌和胞内绵羊布鲁氏菌的差异表达基因(DEGs)。进行了基因本体论(GO)术语富集和京都基因与基因组百科全书(KEGG)通路分析,并采用定量逆转录聚合酶链反应(qRT-PCR)验证了前 10 个上调和下调的 DEGs。结果显示,有 212 个基因差异表达,包括 68 个上调和 144 个下调基因,这些基因主要富集在与生物学过程、细胞成分和分子功能相关的 30 个 GO 术语中。KEGG 分析表明,DEGs 富集在缺氧诱导因子 1(HIF-1)信号通路、丝裂原激活蛋白激酶(MAPK)信号通路、β-丙氨酸代谢和群体感应通路。BME_RS01160、BME_RS04270、BME_RS08185、BME_RS12880、BME_RS25875、predicted_RNA865 和 predicted_RNA953 与转录组测序数据一致。因此,本研究结果不仅揭示了绵羊布鲁氏菌在巨噬细胞免疫系统中的胞内寄生现象,还有助于理解慢性绵羊布鲁氏菌感染的机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdca/7235551/b1a3b7afbc2b/10753_2020_1239_Fig1_HTML.jpg

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