Garcia-Hernandez Vicky, Neumann Philipp-Alexander, Koch Stefan, Lyons Renae, Nusrat Asma, Parkos Charles A
Department of Pathology, University of Michigan.
Department of Surgery, Klinikum Rechts der Isar, Technische Universität München.
J Vis Exp. 2021 Feb 14(168). doi: 10.3791/62135.
Murine colitis models are tools that are extensively employed in studies focused on understanding the pathobiology of inflammatory intestinal disorders. However, robust standards for objective and reproducible quantification of disease severity remain to be defined. Most colitis analysis methods rely on limited histological scoring of small segments of intestine, leading to partial or biased analyses. Here, we combine high-resolution image acquisition and longitudinal analysis of the entire colon to quantify intestinal injury and ulceration in the dextran sodium sulfate (DSS) induced model of murine colitis. This protocol allows for the generation of objective and reproducible results without extensive user training. Here, we provide comprehensive details on sample preparation and image analysis using examples of data from DSS induced colitis. This method can be easily adapted to other models of murine colitis that have significant inflammation associated with mucosal injury. We demonstrate that the fraction of inflamed/injured and eroded/ulcerated mucosa relative to the complete length of the colon closely parallels clinical findings such as weight loss amid DSS-induced disease progression. This histological protocol provides a reliable time and cost-effective aid to standardize analyses of disease activity in an unbiased way in DSS colitis experiments.
小鼠结肠炎模型是广泛应用于专注于理解炎症性肠病病理生物学研究的工具。然而，对于疾病严重程度进行客观且可重复量化的稳健标准仍有待确定。大多数结肠炎分析方法依赖于对小肠小段进行有限的组织学评分，导致分析不全面或有偏差。在此，我们结合高分辨率图像采集和对整个结肠的纵向分析，以量化葡聚糖硫酸钠（DSS）诱导的小鼠结肠炎模型中的肠道损伤和溃疡。该方案无需大量用户培训即可产生客观且可重复的结果。在此，我们使用来自DSS诱导结肠炎的数据示例，提供关于样本制备和图像分析的全面详细信息。此方法可轻松适用于其他与黏膜损伤相关的显著炎症的小鼠结肠炎模型。我们证明，相对于结肠全长，发炎/损伤和糜烂/溃疡黏膜的比例与诸如DSS诱导疾病进展期间体重减轻等临床发现密切平行。这种组织学方案为在DSS结肠炎实验中以无偏差方式标准化疾病活动分析提供了可靠、省时且经济高效的辅助手段。
