Zhang Jing, Zhao Chunping, Yao Min, Qi Jing, Tan Ya, Shi Kaizhi, Wang Jing, Zhou Sixuan, Li Zhixin
Institute of Animal Husbandry and Veterinary Science, Guizhou Academy of Agricultural Sciences, Guiyang 550002, China.
Inspection and Testing Department, Guizhou Testing Center for Livestock and Poultry Germplasm, Guiyang 550002, China.
J Anim Sci Technol. 2024 Jul;66(4):663-681. doi: 10.5187/jast.2023.e46. Epub 2024 Jul 31.
Co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) and (HPS) has severely restricted the healthy development of pig breeding. Exploring disease resistance of non-coding RNAs in pigs co-infected with PRRSV and HPS is therefore critical to complement and elucidate the molecular mechanisms of disease resistance in Kele piglets and to innovate the use of local pig germplasm resources in China. RNA-seq of lungs from Kele piglets with single-infection of PRRSV or HPS and co-infection of both pathogens was performed. Two hundred and twenty-five differentially expressed long non-coding RNAs (DElncRNAs) and 30 DEmicroRNAs (DEmiRNAs) were identified and characterized in the PRRSV and HPS co-infection (PRRSV-HPS) group. Compared with the single-infection groups, 146 unique DElncRNAs, 17 unique DEmiRNAs, and 206 target differentially expressed genes (DEGs) were identified in the PRRSV-HPS group. The expression patterns of 20 DEmiRNAs and DElncRNAs confirmed by real-time quantitative polymerase chain reaction (RT-qPCR) were consistent with those determined by high-throughput sequencing. In the PRRSV-HPS group, the target DEGs were enriched in eight immune Gene Ontology terms relating to two unique DEmiRNAs and 16 DElncRNAs, and the unique target DEGs participated the host immune response to pathogens infection by affecting 15 immune-related Kyoto Encyclopedia of Genes and Genomes enrichment pathways. Notably, competitive endogenous RNA (ceRNA) networks of different groups were constructed, and the miRNA was validated as a potential regulatory factor to regulate and genes to achieve innate antiviral effects and inhibit pulmonary fibrosis by dual-luciferase reporter assays. These results provided insight into further study on the molecular mechanisms of resistance to PRRSV and HPS co-infection in Kele piglets.
猪繁殖与呼吸综合征病毒（PRRSV）和猪肺炎支原体（HPS）的共同感染严重制约了养猪业的健康发展。因此，探索PRRSV和HPS共同感染猪中非编码RNA的抗病性，对于补充和阐明柯乐仔猪抗病的分子机制以及创新中国地方猪种质资源的利用至关重要。对单感染PRRSV或HPS以及两种病原体共同感染的柯乐仔猪的肺组织进行了RNA测序。在PRRSV和HPS共同感染（PRRSV-HPS）组中鉴定并表征了225个差异表达的长链非编码RNA（DElncRNAs）和30个差异表达的微小RNA（DEmiRNAs）。与单感染组相比，PRRSV-HPS组中鉴定出146个独特的DElncRNAs、17个独特的DEmiRNAs和206个靶标差异表达基因（DEGs）。通过实时定量聚合酶链反应（RT-qPCR）验证的20个DEmiRNAs和DElncRNAs的表达模式与高通量测序确定的模式一致。在PRRSV-HPS组中，靶标DEGs富集在与两个独特的DEmiRNAs和16个DElncRNAs相关的八个免疫基因本体学术语中，并且独特的靶标DEGs通过影响15个免疫相关的京都基因与基因组百科全书富集途径参与宿主对病原体感染的免疫反应。值得注意的是，构建了不同组的竞争性内源RNA（ceRNA）网络，并通过双荧光素酶报告基因测定法验证了该miRNA作为潜在调节因子来调节和基因以实现先天性抗病毒作用并抑制肺纤维化。这些结果为进一步研究柯乐仔猪对PRRSV和HPS共同感染的抗性分子机制提供了见解。
