创伤性脑损伤小鼠模型肠通透性动力学

Q1 Agricultural and Biological Sciences Current protocols in mouse biology Pub Date : 2020-12-02 DOI:10.1002/cpmo.86

Jessica L. Weaver

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引用次数: 1

摘要

创伤性脑损伤(TBI)是创伤患者发病和死亡的主要原因。肠通透性增加在TBI的炎症过程中起重要作用，预防这种通透性改变的治疗可能改善TBI患者的预后。已经开发了不同的动物模型来测试渗透性变化，但对于脑外伤后何时测试渗透性尚无一致意见。在这里，我们描述了一种创建TBI小鼠模型和测量肠通透性的方法。我们还详细介绍了脑外伤后不同时间点的渗透率测量，以帮助指导未来的实验设计。脑外伤采用可控皮质冲击模型，皮质冲击器设置为速度6 m/s，深度3 mm，停留时间0.2 s，尖端尺寸3 mm，以产生严重的脑外伤。在脑损伤后2、4、6和24小时，通过移除末端回肠，捆绑末端，在管腔中填充fitc标记的葡聚糖，然后使用荧光板读取器测量随时间推移有多少葡聚糖进入周围的溶液池，测量通透性。我们的研究结果表明，渗透率峰值出现在TBI后4 ~ 6小时。我们建议未来的实验在TBI后4至6小时进行渗透率测量，以利用这一渗透率峰值。©2020 Wiley期刊公司。基本方案:小鼠CCI创伤性脑损伤模型和肠通透性测量

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The Kinetics of Intestinal Permeability in a Mouse Model of Traumatic Brain Injury

Traumatic brain injury (TBI) is a leading cause of morbidity and mortality among trauma patients. Increased intestinal permeability plays an important role in the inflammatory process that accompanies TBI, and therapies that prevent this permeability change may improve outcomes in TBI patients. Different animal models have been developed to test permeability changes, but there has been no agreement on when permeability should be tested after TBI. Here, we describe a method for creating the TBI mouse model and for measuring intestinal permeability. We also detail our permeability measurements at different time points after TBI to help guide future experimental design. The TBI is made using a controlled cortical impact model with the cortical impactor set to speed 6 m/s, depth 3 mm, dwell time 0.2 s, and tip size 3 mm to produce a severe TBI. Permeability is measured at 2, 4, 6, and 24 hr after TBI by removing a piece of terminal ileum, tying the ends, filling the lumen with FITC-labeled dextran, and then measuring how much of the dextran moves into the surrounding solution bath over time using a fluorescent plate reader. Our results show that peak permeability occurs between 4 and 6 hr after TBI. We recommend that future experiments incorporate permeability measurements 4 to 6 hr after TBI in order to take advantage of this peak permeability. © 2020 Wiley Periodicals LLC.

Basic Protocol: Mouse CCI traumatic brain injury model and intestinal permeability measurement

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来源期刊

Current protocols in mouse biology Agricultural and Biological Sciences-Animal Science and Zoology

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet, all too often, nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. The aim of Current Protocols in Mouse Biology is to provide the clearest, most detailed and reliable step-by-step instructions for protocols involving the use of mice in biomedical research. Written by experts in the field and extensively edited to our exacting standards, the protocols include all of the information necessary to complete an experiment in the laboratory—introduction, materials lists with supplier information, detailed step-by-step procedures with helpful annotations, recipes for reagents and solutions, illustrative figures and information-packed tables. Each article also provides invaluable discussions of background information, applications of the methods, important assumptions, key parameters, time considerations, and tips to help avoid common pitfalls and troubleshoot experiments. Furthermore, Current Protocols in Mouse Biology content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Mouse Biology brings together resources in mouse biology and genetics and provides a mouse protocol resource that covers all aspects of mouse biology. Current Protocols in Mouse Biology also permits optimization of mouse model usage, which is significantly impacted by both cost and ethical constraints. Optimal and standardized mouse protocols ultimately reduce experimental variability and reduce the number of animals used in mouse experiments.