产耶尔森杆菌素大肠杆菌诱导小鼠十二指肠炎症的研究

doi:10.16431/j.cnki.1671-7236.2023.11.036

Abstract

Abstract: 【Objective】 The aim of this study was to investigate the effect of yersiniabactin (Ybt) on duodenum inflammation induced by Escherichia coli (E.coli) in mice, and provide new theoretical basis for the mechanism of intestinal inflammation induced by E.coli from pigs.【Method】 The R package was used to mine the gene expression omnibus (GEO) datasets of E.coli infection cells for differential gene volcano plot, heatmap enrichment, and KEGG pathway analysis.Molecular docking software was used to predict the binding of Ybt with Toll-like receptor 4 (TLR4).Mice injected with lipopolysaccharide (LPS) were used as positive control, and mice were infected with E.coli ZB-1 and previously constructed Ybt-deficient strain ZB-1ΔHPI.Duodenum damage after infection was observed by HE staining.Real-time quantitative PCR was used to detect mRNA levels of TLR4, nuclear factor kappa-B (NF-κB), interleukin 18 (IL-18), and IL-1β.ELISA was performed to measure the secretion levels of pro-IL-18, pro-IL-1β, IL-18 and IL-1β in the intestines.Immunohistochemistry (IHC) staining was used to observe the expression and localization of inflammatory factors IL-18 and IL-1β in the intestines.【Result】 Volcano plot and heatmap analysis of public databases showed significant upregulation of TLR4, NF-κB, IL-1β and chemokines (CXCL1, CXCL3, CXCL5, and so on) in response to E.coli infection (P<0.05).KEGG enrichment analysis revealed the involvement of TLR4/NF-κB pathway in E.coli infection.Additionally, molecular docking showed that Ybt could bind to TLR4.HE staining demonstrated that E.coli infection with Ybt induction resulted in more severe duodenum inflammation and damage in mice.Real-time quantitative PCR results showed that mRNA levels of TLR4, NF-κB, IL-18, and IL-1β were significantly higher in ZB-1 group compared to ZB-1ΔHPI group (P<0.05).ELISA results showed that ZB-1 group significantly promoted the release of pro-IL-Iβ, IL-Iβ, pro-IL-18 and IL-18 compared to ZB-1ΔHPI group (P<0.05).Furthermore, IHC results showed abundant expression of IL-18 and IL-1β in duodenum villous epithelial cells, and Ybt producing E.coli infection significantly enhanced this result (P<0.05).【Conclusion】 Ybt exhibited a strong pro-inflammatory effect and promotes E.coli-induced duodenum inflammation in mice through the TLR4/NF-κB pathway.These finding providde new theoretical evidence for the mechanism of duodenum inflammation induced by porcine-derived E.coli.

Key words: Escherichia coli; yersiniabactin; TLR4/NF-κB pathway; duodenum inflammation

CLC Number:

S852.61

WANG Hao, JING Linxi, XIAO Jinlong, SHEN Jue, ZHAO Jingang, WANG Shuai, LIU Gen, ZHAO Ru, XIAO Peng, GAO Hong. Study on the Induction of Duodenum Inflammation in Mice by Yersiniabactin Producing Escherichia coli[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4693-4702.

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Study on the Induction of Duodenum Inflammation in Mice by Yersiniabactin Producing Escherichia coli

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