Oxazolone-Induced Enterocolitis in Zebrafish Depends on the Composition of the Intestinal Microbiota
Oxazolone-Induced Enterocolitis in Zebrafish Depends on the Composition of the Intestinal Microbiota
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DOI：
10.1053/j.gastro.2009.07.069
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发表时间：
2009-11-01
期刊：
影响因子：
29.4
通讯作者：
Nieuwenhuis, Edward E. S.
中科院分区：
文献类型：
--
作者：
Brugman, Sylvia;Liu, Kit-Yeng;Nieuwenhuis, Edward E. S.
背景与目的：炎症性肠病的发病机制涉及在遗传易感宿主中对肠道细菌的功能失调的粘膜免疫应答。宿主细胞和细菌之间的相互作用是复杂的，这使得评估它们对肠道病理学的相对贡献成为一个挑战。我们开发了一种斑马鱼小肠结肠炎模型来研究这些相互作用。方法：小肠结肠炎诱导的半抗原恶唑酮在成年野生型和髓过氧化物酶报告基因转基因斑马鱼在存在或不存在抗生素的直肠内给药。通过组织学和流式细胞术分析以及定量实时聚合酶链反应的细胞因子分析来评估肠道炎症。通过16 SrRNA测序评估抗生素给药后肠道微生物群组成的变化，并通过在非选择性培养基上培养（菌落形成单位）定量细菌负荷。结果：在斑马鱼中，恶唑酮诱导的小肠结肠炎的浸润和严重程度受到微生物群组成的影响。炎症的特征在于粒细胞内流;上皮损伤;杯状细胞耗竭;以及白细胞介素-1 β、肿瘤坏死因子-α和白细胞介素-10的表达增加。给予万古霉素的斑马鱼的细菌种群以梭杆菌属为主，并降低了小肠结肠炎评分、肠道损伤以及浸润中性粒细胞和嗜酸性粒细胞的百分比。相比之下，斑马鱼给予硫酸粘杆菌素具有优势的protecibacteria和减少嗜酸性粒细胞和淋巴细胞浸润，但小肠结肠炎评分没有减少。结论：在患有恶唑酮诱导的小肠结肠炎的斑马鱼中，肠道微生物群的组分影响肠道浸润的严重程度和组成。
BACKGROUND & AIMS: The pathogenesis of inflammatory bowel disease involves dysfunctional mucosal immune responses to commensal bacteria in genetically predisposed hosts. Interactions between host cells and bacteria are complicated, making it a challenge to assess their relative contribution to intestinal pathology. We developed a zebrafish model of enterocolitis to Study these interactions. METHODS: Enterocolitis was induced by intrarectal administration of the hapten oxazolone in adult wild-type and myeloperoxidase-reporter transgenic zebrafish in the presence or absence of antibiotics. Intestinal inflammation was evaluated by histological and flow cytometry analyses and cytokine profiling with quantitative real-time polymerase chain reaction. Changes in the composition of the intestinal microbiota following antibiotic administration were assessed by 16SrRNA sequencing and bacterial load was quantified by culture on nonselective media (colony-forming units). RESULTS: In zebrafish, the infiltrate and severity of oxazolone-induced enterocolitis are influenced by the composition of the microbiota. Inflammation is characterized by granulocyte influx; epithelial damage; goblet cell depletion; and increased expression of interleukin-1 beta, tumor necrosis factor-alpha, and interleukin-10. Zebrafish given vancomycin had bacterial populations dominated by Fusobacteria and reduced enterocolitis scores, intestinal damage, and percentages of infiltrating neutrophils and eosinophils. In contrast, zebrafish given colistin sulphate had a predominance of protecibacteria and reduced eosinophil and lymphocyte infiltration, but enterocolitis scores were not reduced. CONCLUSIONS: In zebrafish with oxazolone-induced enterocolitis, components of the intestinal microbiota affect the severity and composition of the intestinal infiltrate.