In Vivo Analysis of Gut Function and Disease Changes in a Zebrafish Larvae Model of Inflammatory Bowel Disease: A Feasibility Study
In Vivo Analysis of Gut Function and Disease Changes in a Zebrafish Larvae Model of Inflammatory Bowel Disease: A Feasibility Study
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DOI：
10.1002/ibd.21200
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发表时间：
2010-07-01
影响因子：
4.9
通讯作者：
Goldsmith, Paul
中科院分区：
文献类型：
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作者：
Fleming, Angeleen;Jankowski, Janusz;Goldsmith, Paul
背景：本研究的目的是建立一种斑马鱼幼虫炎症性肠病（IBD）模型，以及一种快速评估体内肠道形态和功能的方法，从而实现中通量化合物筛选。方法：在96孔板中对受精后38天的斑马鱼幼虫进行检测。摄入荧光染料后，在体内用荧光成像观察肠道形态和蠕动。通过在井内培养基中添加2,4,6-三硝基苯磺酸（TNBS）诱导IBD。在体内用荧光成像评估病理，死后用组织学、免疫组织化学和电子显微镜评估病理。治疗性化合物与TNBS联合给药进行评价。结果：设计了一种利用斑马鱼活体幼虫荧光成像研究肠道结构和蠕动的新方法。在整个肠道中观察到与结肠炎一致的肠道结构的原型变化。杯状细胞数量和肿瘤坏死因子α (tnf - α)抗体染色的显著变化用于量化疾病严重程度和抢救。强的松龙联合5-氨基水杨酸治疗可改善病情变化。评估候选治疗化合物（NOS抑制剂、沙利度胺和孤雌醇内酯），并观察到使用单一生化测量（tnf - α染色）评估疗效与评估整个疾病状态之间的分离。结论：与人类疾病状态相关的肠道生理病理可以在斑马鱼幼体中快速建模。该模型适用于中等通量化学筛选，并可进行遗传操作，因此为胃肠道疾病的研究提供了一种强大的新型前哺乳动物辅助工具。
Background: The aim of this study was to develop a model of inflammatory bowel disease (IBD) in zebrafish larvae, together with a method for the rapid assessment of gut morphology and function in vivo thereby enabling medium-throughput compound screening.Methods: Assays were performed using larval zebrafish from 3 8 days postfertilization (d.p.f.) in 96-well plates. Gut morphology and peristalsis were observed in vivo using fluorescent imaging following ingestion of fluorescent dyes. IBD was induced by addition of 2,4,6-trinitrobenzenesulfonic acid (TNBS) to the medium within the well. Pathology was assessed in vivo using fluorescent imaging and postmortem by histology, immunohistochemistry, and electron microscopy. Therapeutic compounds were evaluated by coadministration with TNBS.Results: A novel method of investigating gut architecture and peristalsis was devised using fluorescent imaging of live zebrafish larvae. Archetypal changes in gut architecture consistent with colitis were observed throughout the gut. Significant changes in goblet cell number and tumor necrosis factor alpha (TNF-alpha) antibody staining were used to quantify disease severity and rescue. Prednisolone and 5-amino salicylic acid treatment ameliorated the disease changes. Candidate therapeutic compounds (NOS inhibitors, thalidomide, and parthenolide) were assessed and a dissociation was observed between efficacy assessed using a single biochemical measure (TNF-alpha staining) versus an assessment of the entire disease state.Conclusions: Gut physiology and pathology relevant to human disease state can be rapidly modeled in zebrafish larvae. The model is suitable for medium-throughput chemical screens and is amenable to genetic manipulation, hence offers a powerful novel premammalian adjunct to the study of gastrointestinal disease.