基于高通量测序分析肺炎支原体感染姜曲海猪肺组织miRNA表达谱

为探明姜曲海猪感染肺炎支原体后肺组织miRNA(microRNA)表达谱和分子机制,选取50日龄姜曲海猪为实验猪,随机分成感染组和对照组,人工感染肺炎支原体28 d后,采集肺部组织,进行高通量miRNA测序,采用生物信息学软件进行miRNA鉴定和靶基因预测。结果显示:感染组和对照组的肺组织分别筛选到14 265 786条和14 000 588条小RNA纯净序列(clean reads)。与对照组相比,感染组中筛选到73个显著差异表达的miRNAs,其中39个表达上调,34个表达下调,从中随机选取4个miRNAs进行定量PCR(quantitative real time PCR,qRT-PCR)验证,感染组和对照组的表达水平与测序结果基本一致。73个差异表达miRNAs预测到1 685个靶基因和4 220个靶位点,靶基因预测筛选到8个与免疫调控相关的miRNAs。靶基因GO(gene ontology)分析显示,miRNA广泛参与生物过程、细胞组成和分子功能的调控。靶基因KEGG(Kyoto encyclopedia of genes and genomes)分析显示,miRNA参与调控细胞凋亡、ECM受体相互作用、粘附斑通路等信号通路。本研究通过高通量测序获得姜曲海猪感染肺炎支原体后肺组织miRNA表达谱,通过生物信息学分析筛选到与免疫调控相关的miRNAs和信号通路,为进一步阐明姜曲海猪的肺炎支原体感染机制奠定了基础。

MicroRNA sequencing and analysis of porcine lung of Jiangquhai pig response to Mycoplasma hyopneumoniae infection

The purpose of the study was to explore the expression profile and mechanism of miRNA of porcine lung response to Mycoplasma hyopneumoniae infection. In this experiment, Jiangquhai pigs were divided into the infected group and the control group, and the infected group was inoculated with Mycoplasma hyopneumoniae. At the 28th day after inoculation, all pigs were slaughtered and lung tissues were collected for high-throughput microRNA sequencing. The differentially expressed miRNAs were screened and the related target genes were predicted using several prediction softwares. The results showed that a total of 14 265 786 and 14 000 588 clean reads were obtained from the infected group and the control group respectively. Compared with the control group, 73 significant differentially expressed miRNAs were obtained from infected group, 39 and 34 miRNAs were up-regulated and down-regulated. Among them, 4 miRNAs were randomly selected and verified by qRT-PCR, the expression pattern were consistent with the sequencing results. 1 685 target genes and 4 220 target sites for all differentially expressed miRNAs were predicted. 8 miRNAs were related to immunoregulation. The predicted target genes were analyzed by GO, which showed that these miRNA were widely involved in the regulation of biological processes, cellular components and molecular functions cells. KEGG pathway analysis showed that the miRNAs were involved in the regulation of apoptosis, ECM-receptor interaction, focal adhesion and other signaling pathways. In this study, we obtained the miRNA expression profile of porcine lung of Jiangquhai pig response to Mycoplasma hyopneumoniae infection, and immunity-associated miRNAs and signal pathways were obtained by bioinformatics analysis, the results were helpful for understanding the mechanism of Mycoplasma hyopneumoniae infection in Jiangquhai pigs.