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一种简单的肠道模型,用于研究大肠杆菌与盲肠黏液中肠道共生菌群的相互作用。

A Simple Gut Model for Studying the Interaction between Escherichia coli and the Intestinal Commensal Microbiota in Cecal Mucus.

机构信息

Department of Cell and Molecular Biology, University of Rhode Island, Kingston, Rhode Island, USA.

Department of Microbiological Surveillance and Research, Statens Serum Institut, Copenhagen, Denmark.

出版信息

Appl Environ Microbiol. 2018 Nov 30;84(24). doi: 10.1128/AEM.02166-18. Print 2018 Dec 15.

Abstract

A novel gut model was developed to better understand the interactions between and the mouse cecal mucus commensal microbiota. The gut model is simple and inexpensive while providing an environment that largely replicates the nonadherent mucus layer of the mouse cecum. 16S rRNA gene profiling of the cecal microbial communities of streptomycin-treated mice colonized with MG1655 or Nissle 1917 and the gut model confirmed that the gut model properly reflected the community structure of the mouse intestine. Furthermore, the results from the gut model mimic the results of published competitive colonization experiments. The gut model is initiated by the colonization of streptomycin-treated mice, and then the community is serially transferred in microcentrifuge tubes in an anaerobic environment generated in anaerobe jars. The nutritional makeup of the cecum is simulated in the gut model by using a medium consisting of porcine mucin, mouse cecal mucus, HEPES-Hanks buffer (pH 7.2), Cleland's reagent, and agarose. Agarose was found to be essential for maintaining the stability of the microbial community in the gut model. The outcome of competitions between strains in the gut model is readily explained by the "restaurant hypothesis" of intestinal colonization. This simple model system potentially can be used to more fully understand how different members of the microbiota interact physically and metabolically during the colonization of the intestinal mucus layer. Both commensal and pathogenic strains of appear to colonize the mammalian intestine by interacting physically and metabolically with other members of the microbiota in the mucus layer that overlays the cecal and colonic epithelium. However, the use of animal models and the complexity of the mammalian gut make it difficult to isolate experimental variables that might dictate the interactions between and other members of the microbiota, such as those that are critical for successful colonization. Here, we describe a simple and relatively inexpensive gut model that largely mimics conditions and therefore can facilitate the manipulation of experimental variables for studying the interactions of with the intestinal microbiota.

摘要

开发了一种新型肠道模型,以更好地了解 与小鼠盲肠粘液共生菌群之间的相互作用。该肠道模型简单且经济实惠,同时提供了一个很大程度上复制小鼠盲肠非粘附粘液层的环境。对链霉素处理的小鼠盲肠微生物群落进行 16S rRNA 基因谱分析,发现定植 MG1655 或 Nissle 1917 的 小鼠的肠道模型证实了该肠道模型正确反映了小鼠肠道的群落结构。此外,肠道模型的结果模拟了已发表的 竞争定植实验的结果。肠道模型通过定植链霉素处理的小鼠开始,然后在厌氧罐中产生的厌氧环境中,在微量离心管中连续转移群落。通过使用由猪粘蛋白、小鼠盲肠粘液、HEPES-Hanks 缓冲液(pH7.2)、Cleland 试剂和琼脂糖组成的培养基,在肠道模型中模拟盲肠的营养组成。发现琼脂糖对于维持肠道模型中微生物群落的稳定性至关重要。在肠道模型中, 菌株之间的竞争结果可以很容易地用肠道定植的“餐馆假说”来解释。这个简单的模型系统有可能被用来更充分地了解微生物群落的不同成员在定植肠道粘液层时如何在物理和代谢上相互作用。共生和致病的 菌株似乎通过与覆盖盲肠和结肠上皮的粘液层中的其他微生物成员在物理和代谢上相互作用来定植哺乳动物肠道。然而,使用动物模型和哺乳动物肠道的复杂性使得很难分离可能决定 与其他微生物群落之间相互作用的实验变量,例如那些对成功定植至关重要的变量。在这里,我们描述了一种简单且相对便宜的 肠道模型,该模型在很大程度上模拟了 条件,因此可以方便地操纵实验变量,以研究 与肠道微生物群的相互作用。

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