LncRNA-NEAT1介导的RNA核滞留效应在梅山猪巨噬细胞感染肺炎支原体炎症反应调控中的作用

Swine mycoplasma pneumonia, commonly known as swine asthma, is a high incidence disease for the excellent domestic breeding of meishan pig, which resulted in seriously affecting the exploitation and utilization of the meishan pig. However, the molecular mechanisms of the interaction between this disease and meishan pig is unclear. Inflammatory response mediated by innate immune system plays a key role in the host to resist infection by mycoplasma pneumoniae. Understanding the precise release of inflammatory cytokines will be in favor of understanding the molecular mechanisms of the host against mycoplasma pneumonia. NEAT1is a newly discovered lncRNA, which plays an important role in the regulation of inflammation. In this study, meishan porcine alveolar macrophage was used as a mycoplasma pneumoniae infection model. Using this model, RNA interference and proteomics and EGFP marker were used to identify the target gene for NEAT1 through RNA nuclear retention and explore the interaction regulation between target gene and inflammatory factors. This study contributed to a better understanding of the precise regulation mechanism of inflammatory response in meishan pig during mycoplasma pneumonia infection and provide a theoretical basis for the effective prevention and control measures and meanwhile develop the basic knowledge of natural immunity signal transduction and the fine adjustment, which has important academic and production value.

猪支原体肺炎俗称猪气喘病，是国内地方优秀品种梅山猪高发的一种疾病，已严重影响到该品种的开发与利用，然而梅山猪与该疾病互作的分子机制尚不清晰。天然免疫系统介导的炎症反应在宿主抵御肺炎支原体感染中起着关键作用，了解炎症因子的精确释放有利于认识宿主抵御支原体肺炎的分子机制。 NEAT1是新近发现的一种长链非编码RNA，在炎症反应中的调控中发挥着重要作用。本研究以梅山猪肺泡巨噬细胞为肺炎支原体感染模型，通过RNA干扰、蛋白组组学和EGFP标记实验来鉴定NEAT1通过RNA核滞留效应调控的靶基因以及探索这些靶基因与炎症因子的调控互作关系。本研究的开展有助于深入理解梅山猪感染肺炎支原体后炎症反应的精确调控机制，为提出有效防控措施提供了理论基础，同时也丰富和发展了天然免疫信号转导和精细调节的基础知识，具有重要的学术价值和生产应用价值。

天然免疫系统介导的炎症反应在宿主抵御肺炎支原体感染中起着关键作用， 了解炎症因子的精确释放有利于认识宿主抵御支原体肺炎的分子机制。NEAT1是新近发现的一种长链非编码RNA，在炎症反应中的调控中发挥着重要作用。本研究探讨了NEAT1的表达特征及其表达量变化与炎症因子的关系，在此基础上分析了猪单核巨噬细胞受支原体脂蛋白刺激后的转录组学以及lncRNAs的鉴定与特征。利用生物信息学鉴定了猪全基因组范围内蛋白编码基因3'UTR中IRPre1元件，并对NEAT1推测的靶基因的IRPre1元件进行了功能分析。结果表明NEAT1表达在支原体感染猪巨噬细胞后呈现时空依赖性，其正调控炎症因子CXCL8和IL6。猪单核巨噬细胞受支原体脂蛋白刺激后的差异蛋白编码基因主要参与TNF信号通路、细胞因子-受体互作、Toll样受体通路以及Jak-STAT信号通路等；而差异的lncRNAs也趋向位于与炎症、免疫相关的蛋白编码基因附近。通过生物信息学鉴定了1094个含有IRPre1元件的蛋白编码基因，这些候选基因主要参与了缬草碱、亮氨酸和异亮氨酸降解途径、TNF信号通路、T细胞受体信号通路、RIG-I样受体信号通路，同时NEAT1推测的靶基因BAG4的IRPre1元件对绿色荧光蛋白的翻译起到阻滞作用。本研究初步揭示了NEAT1对炎症因子的作用，解析了其靶基因IRPre1元件的功能。研究结果为进一步阐明NEAT1在猪与支原体诱导的炎症反应中的调控机理提供了基础。