基于猪小肠上皮细胞炎症模型分析产肠毒素大肠杆菌诱导的转录组表达变化

doi:10.16431/j.cnki.1671-7236.2022.12.004

Abstract

Abstract: 【Objective】 This study was aimed to analyze the key host-regulated genes and related molecular pathways in the inflammatory response induced by enterotoxigenic Escherichia coli (ETEC) in porcine small intestinal epithelial cells (IPEC-J2), in order to provide theoretical basis for the investigation of inflammatory mechanism induced by ETEC infection.【Method】 The IPEC-J2 cells were infected with ETEC bacterial solution with a multiplicity of infection (MOI) of 100, and the medium supernatant was collected at 18 h post infection.The secretion of proinflammatory factor interleukin 1β (IL-1β) was detected by ELISA method.High throughput transcriptome sequencing was performed by Illumina PE150 sequencing platform.Differentially expressed analysis was performed on the sequencing results by edgeR v 3.12.1 software.GO function and KEGG pathway enrichment analysis were performed on the differentially expressed mRNA obtained by GOseq and KOBAS 2.0 softwares.Five differentially expressed mRNA were randomly selected and verified by Real-time quantitative PCR.【Result】 A model of ETEC infection-induced inflammation in IPEC-J2 cells was successfully established.Compared with control group, a total of 529 differentially expressed mRNA were identified in inflammatory model group, of which 236 were up-regulated and 293 were down-regulated, including immune-related genes such as HSP90AA1, CGNL1, CADPS2 and EHBP1.GO function analysis showed that differentially expressed mRNA after ETEC infection were significantly enriched in cellular component and biological process, such as the intracellular part, cytoplasm, nuclear part, intracellular membrance-bound organelle, membrance-bound orgabelle and cellular process.KEGG pathway analysis showed that most of differentially expressed genes after ETEC infection were annotated as disease-related pathways as well as immune-related signaling pathways such as Toll-like receptor signaling pathway, mTOR signaling pathway, cell cycle and adhesion junctions.Real-time quantitative PCR was used to validate the expression of five randomly selected differentially expressed mRNA, the results were consistent with the trend of differentially expressed mRNA in the sequencing result, which indicated that the sequencing results were reliable.【Conclusion】 HSP90AA1, CGNL1, CADPS2 and EHBP1 genes might play a key role in ETEC adhesion to IPEC-J2 cells and induce inflammatory responses, and immunoregulated through Toll-like receptors, mTOR and other signaling pathways.The results provided a reference for further understanding the molecular mechanisms and regulatory networks involved in the inflammatory response induced by ETEC infection.

Key words: enterotoxigenic Escherichia coli (ETEC); porcine small intestinal epithelial cells (IPEC-J2); inflammation model; transcriptome

CLC Number:

S852.61⁺2

ZHU Yue, LIU Yingying, LI Yinghui, LI Pei, ZHU Kaiqing, JIA Tong, LI Yan. Analysis of Enterotoxigenic Escherichia coli Induced Transcriptomic Changes of Porcine Intestinal Epithelial Cell Based on the Inflammatory Model[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4545-4553.

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Analysis of Enterotoxigenic Escherichia coli Induced Transcriptomic Changes of Porcine Intestinal Epithelial Cell Based on the Inflammatory Model

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