人支气管上皮细胞感染鹦鹉热衣原体后 microRNA 表达谱分析。
Analysis of microRNA expression profiles in human bronchial epithelial cells infected by Chlamydia psittaci.
机构信息
Department of public health laboratory sciences, College of Public Health, University of South China, Hengyang, China; Key Laboratory of Hengyang for Health Hazard Factors Inspection and Quarantine, Hengyang, China.
Department of public health laboratory sciences, College of Public Health, University of South China, Hengyang, China; Key Laboratory of Hengyang for Health Hazard Factors Inspection and Quarantine, Hengyang, China; Department of Infection Control, The First People's Hospital of Yunnan Province, Kunming, China.
出版信息
Microb Pathog. 2021 May;154:104837. doi: 10.1016/j.micpath.2021.104837. Epub 2021 Mar 6.
BACKGROUND
Chlamydia psittaci is a pathogen of birds that can cause zoonotic disease in mammals including pneumonia in humans. MicroRNAs (miRNAs) are a class of small non-coding RNA fragments with a length of about 22 nt, which play an important role in regulating gene expression after transcription. Chlamydia infection can cause changes in host cell miRNA expression, but the potential biological function of miRNAs in C. psittaci infection and pathogenesis is not well understood.
METHODS
Small RNA sequencing (sRNA-Seq) technology was used to characterise miRNA expression in human bronchial epithelial (HBE) cells after C. psittaci infection, and differentially expressed miRNAs were identified. Candidate target genes for these miRNAs were then functionally annotated by Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The sRNA-Seq results were partially validated by quantitative real time polymerase chain reaction (qRT-PCR) and miRNA-target networks were constructed using visualization software.
RESULTS
We identified 151 differentially expressed miRNAs (46 known miRNAs and 105 novel miRNAs) in C. psittaci-infected HBE cells, of which 140 were upregulated and 11 were downregulated. Of these, 17 known miRNAs were significantly upregulated and two were downregulated using P < 0.05 and |logFoldChange|>1.5 as threshold criteria. GO enrichment results showed that the predicted targets of these differentially expressed miRNAs were mainly involved in transcriptional regulation and ATP binding. KEGG pathway analysis suggested that the candidate target genes were involved in several important signaling pathways such as MAPK, ErbB, cGMP-PKG, cAMP, mTOR, GNRH, oxytocin, PI3K-Akt and AMPK, which are primarily related to biological processes such as transcription and signal transduction. The qRT-PCR results for miR-2116-3p, miR-3195, miR-663a, miR-10401-5p, miR-124-3p, miR-184, miR-744-5p and hsa-miR-514b-5p were consistent with the sRNA-Seq data.
CONCLUSIONS
A large amount of miRNA expression profile data relating to C. psittaci infection was obtained, which provides a useful experimental and theoretical basis for further understanding the pathogenic mechanisms of C. psittaci infection.
背景
鹦鹉热衣原体是一种禽类病原体，可引起哺乳动物的人畜共患病，包括人类肺炎。微小 RNA（miRNA）是一类长度约为 22nt 的小非编码 RNA 片段，在转录后调控基因表达中发挥重要作用。衣原体感染可导致宿主细胞 miRNA 表达发生变化，但 miRNA 在鹦鹉热衣原体感染和发病机制中的潜在生物学功能尚不清楚。
方法
采用小 RNA 测序（sRNA-Seq）技术对鹦鹉热衣原体感染人支气管上皮（HBE）细胞后的 miRNA 表达进行了特征分析，鉴定出差异表达的 miRNA。然后，通过基因本体论（GO）分析和京都基因与基因组百科全书（KEGG）通路分析对这些 miRNA 的候选靶基因进行功能注释。sRNA-Seq 结果部分通过实时定量聚合酶链反应（qRT-PCR）验证，并使用可视化软件构建 miRNA-靶网络。
结果
我们在鹦鹉热衣原体感染的 HBE 细胞中鉴定出 151 个差异表达的 miRNA（46 个已知 miRNA 和 105 个新 miRNA），其中 140 个上调，11 个下调。其中，使用 P<0.05 和 |logFoldChange|>1.5 作为阈值标准，有 17 个已知 miRNA 显著上调，有 2 个下调。GO 富集结果表明，这些差异表达 miRNA 的预测靶标主要参与转录调控和 ATP 结合。KEGG 通路分析表明，候选靶基因参与了 MAPK、ErbB、cGMP-PKG、cAMP、mTOR、GNRH、催产素、PI3K-Akt 和 AMPK 等几个重要信号通路，这些信号通路主要与转录和信号转导等生物学过程有关。miR-2116-3p、miR-3195、miR-663a、miR-10401-5p、miR-124-3p、miR-184、miR-744-5p 和 hsa-miR-514b-5p 的 qRT-PCR 结果与 sRNA-Seq 数据一致。
结论
获得了大量与鹦鹉热衣原体感染相关的 miRNA 表达谱数据，为进一步了解鹦鹉热衣原体感染的发病机制提供了有用的实验和理论依据。
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