Department of Biological and Environmental Sciences and Technologies, University of Salento, Via Monteroni, 73100, Lecce, Italy.
Department of Soil Plant and Food Sciences, University of Bari Aldo Moro, 70126, Bari, Italy.
Inflammation. 2022 Dec;45(6):2477-2497. doi: 10.1007/s10753-022-01706-0. Epub 2022 Jun 22.
Winnie, a mouse carrying a missense mutation in the MUC2 mucin gene, is a valuable model for inflammatory bowel disease (IBD) with signs and symptoms that have multiple similarities with those observed in patients with ulcerative colitis. MUC2 mucin is present in Winnie, but is not firmly compacted in a tight inner layer. Indeed, these mice develop chronic intestinal inflammation due to the primary epithelial defect with signs of mucosal damage, including thickening of muscle and mucosal layers, goblet cell loss, increased intestinal permeability, enhanced susceptibility to luminal inflammation-inducing toxins, and alteration of innervation in the distal colon. In this study, we show that the intestinal environment of the Winnie mouse, genetically determined by MUC2 mutation, selects an intestinal microbial community characterized by specific pro-inflammatory, genotoxic, and metabolic features that could imply a direct involvement in the pathogenesis of chronic intestinal inflammation. We report results obtained by using a variety of in vitro approaches for fecal microbiota functional characterization. These approaches include Caco-2 cell cultures and Caco-2/THP-1 cell co-culture models for evaluation of geno-cytotoxic and pro-inflammatory properties using a panel of 43 marker RNAs assayed by RT-qPCR, and cell-based phenotypic testing for metabolic profiling of the intestinal microbial communities by Biolog EcoPlates. While adding a further step towards understanding the etiopathogenetic mechanisms underlying IBD, the results of this study provide a reliable method for phenotyping gut microbial communities, which can complement their structural characterization by providing novel functional information.
Winnie 是一只携带 MUC2 粘蛋白基因突变的小鼠,是一种具有多种与溃疡性结肠炎患者相似症状和体征的炎症性肠病(IBD)的有价值模型。Winnie 存在 MUC2 粘蛋白,但它没有紧密地紧密地包裹在一个紧密的内层中。事实上,这些小鼠由于主要上皮缺陷而发展为慢性肠道炎症,表现为黏膜损伤的迹象,包括肌肉和黏膜层增厚、杯状细胞丢失、肠道通透性增加、对腔内炎症诱导毒素的易感性增强以及远端结肠的神经支配改变。在这项研究中,我们表明,Winnie 小鼠的肠道环境由 MUC2 突变决定,选择了一种具有特定促炎、遗传毒性和代谢特征的肠道微生物群落,这可能意味着它们直接参与慢性肠道炎症的发病机制。我们报告了使用多种体外方法对粪便微生物群落功能进行特征描述的结果。这些方法包括 Caco-2 细胞培养物和 Caco-2/THP-1 细胞共培养模型,用于使用 43 个标记 RNA 的 RT-qPCR 检测评估遗传毒性和促炎特性,以及细胞表型测试用于通过 Biolog EcoPlates 对肠道微生物群落进行代谢分析。虽然这进一步加深了我们对 IBD 发病机制的理解,但这项研究的结果提供了一种可靠的方法来对肠道微生物群落进行表型分析,这可以通过提供新的功能信息来补充其结构特征描述。