College of Veterinary Medicine, Hebei Agricultural University, Baoding, China.
Front Cell Infect Microbiol. 2022 Aug 25;12:996841. doi: 10.3389/fcimb.2022.996841. eCollection 2022.
Enterotoxigenic (ETEC) is a major cause of bacterial diarrhea in piglets, leading to economic losses in the pig industry. In past decades, long non-coding RNAs (lncRNAs) have shown to be widely involved in the regulation of host immunity in porcine infection diseases. In this study, we explored the lncRNAs associated with ETEC F41 infection in IPEC-J2 cells by high-throughput sequencing and bioinformatic analysis. A total of 10150 novel porcine lncRNAs were identified. There were 161 differentially expressed (DE) lncRNAs associated with ETEC F41 infection, of which 65 DE lncRNAs were up-regulated and 96 DE lncRNAs were down-regulated. Functional and KEGG enrichment analysis of predicted target genes of DE lncRNAs indicated they are enriched in cell growth and inflammation-related pathways, such as endocytosis, focal adhesion, TGF-β signaling pathway, and adherens junctions. We revealed a novel candidate lncRNA MPHOSPH9-OT1 that was up-regulated after ETEC infection. The qRT-PCR validation and ELISA assessment showed the knockdown and overexpression of MPHOSPH9-OT1 resulted in significantly down- and up-regulation of cellular mRNA levels and secreted cytokine levels of CXCL8/IL-8, respectively. Meanwhile, MPHOSPH9-OT1 equilibrium is important to maintain the transepithelial electric resistance value and tight junction protein expression of IPEC-J2 cells. This study provides insights into the functionality of novel porcine lncRNAs in host immune responses to ETEC infection.
肠产毒性大肠杆菌（ETEC）是导致仔猪细菌性腹泻的主要原因，给养猪业造成了经济损失。在过去的几十年中，长链非编码 RNA（lncRNA）已被证明广泛参与宿主感染性疾病的免疫调节。在这项研究中，我们通过高通量测序和生物信息学分析，研究了与 IPEC-J2 细胞 ETEC F41 感染相关的 lncRNA。共鉴定出 10150 个新的猪 lncRNA。有 161 个与 ETEC F41 感染相关的差异表达（DE）lncRNA，其中 65 个 DE lncRNA 上调，96 个 DE lncRNA 下调。预测 DE lncRNA 靶基因的功能和 KEGG 富集分析表明，它们富集在与细胞生长和炎症相关的途径中，如内吞作用、焦点黏附、TGF-β 信号通路和黏着连接。我们揭示了一个新的候选 lncRNA MPHOSPH9-OT1，它在 ETEC 感染后上调。qRT-PCR 验证和 ELISA 评估表明，MPHOSPH9-OT1 的敲低和过表达分别导致细胞 mRNA 水平和分泌细胞因子 CXCL8/IL-8 的水平显著下调和上调。同时，MPHOSPH9-OT1 的平衡对于维持 IPEC-J2 细胞的跨上皮电阻值和紧密连接蛋白表达很重要。这项研究为宿主对 ETEC 感染的免疫反应中新型猪 lncRNA 的功能提供了新的见解。
