Murine model of Hirschsprung-associated enterocolitis. I: Phenotypic characterization with development of a histopathologic grading system
Murine model of Hirschsprung-associated enterocolitis. I: Phenotypic characterization with development of a histopathologic grading system
复制标题
DOI：
10.1016/j.jpedsurg.2009.06.009
复制
发表时间：
2010-03-01
影响因子：
2.4
通讯作者：
Frykman, Philip K.
中科院分区：
文献类型：
--
作者：
Cheng, Zhi;Dhall, Deepti;Frykman, Philip K.
目的：本研究的目的是描述小肠结肠炎的Ednrb-null（Ednrb-/-）小鼠与结肠无神经节细胞症和开发和验证半定量组织病理学分级系统，以评估enterocolitis.Methods：我们分离的Ednrb-/-和对照小鼠（Ednrb+/+）的结肠和回肠标本，并进行组织化学染色（H&E）的组织切片。在建立炎症分级标准后，2名设盲的病理学家在2个不同的场合独立评估近端结肠段的炎症严重程度和深度。计算组间相关系数（ICC）和变异系数（CV），以确定评价者间和评价者内一致性。然后，我们前瞻性地将小肠结肠炎分级系统应用于临床患病的Ednrb-/-小鼠。观察一组Ednrb-/-小鼠，直到它们发展出临床疾病，此时将它们安乐死，并对多个器官匀浆进行细菌培养，并对结肠和小肠进行小肠结肠炎的组织病理学分级。斯皮尔曼的等级相关性比较小肠结肠炎评分与菌血症的水平进行了performed.Results：内部和interrater ICC的组织学评分系统是令人满意的（0.61和0.94，分别），作为内部和interrater CV（18%和9%，分别）。在Ednrb-/-小鼠中，65%发生了菌血症。有菌血症者小肠结肠炎评分显著高于无菌血症者（P <0.01）。Ednrb-/-小鼠，发展菌血症表现出很强的正相关性之间的总小肠结肠炎评分和细菌菌落形成单位的数量在腹腔灌洗，肝脏，肾脏，有氧spleen.Conclusions：Ednrb-/-小鼠与无神经节细胞症发展小肠结肠炎，并具有类似于Hirschsprung相关的小肠结肠炎在人类的功能。我们的分级系统是评估小肠结肠炎的可靠方法。通过进行显微外科拉通，我们现在可以进行对照，假设驱动，机制研究，以评估影响Ednrb-/-小鼠小肠结肠炎的病因因素。(C)2010年爱思唯尔公司All rights reserved.
Purpose: The aim of the study was to characterize enterocolitis in the Ednrb-null (Ednrb-/-) mouse with aganglionosis of the colon and to develop and validate a semiquantitative histopathologic grading system to assess enterocolitis.Methods: We isolated colon and ileal specimens of Ednrb-/- and control mice (Ednrb+/+) and performed histochemical staining (H&E) on tissue sections. After establishing inflammation grading criteria, 2 blinded pathologists independently assessed the severity and depth of inflammation of proximal colon segments on 2 separate occasions. Interclass correlations (ICCs) and coefficient of variation (CV) were calculated to determine interrater and intrarater agreement. We then prospectively applied the enterocolitis grading system to Ednrb-/- mice that became clinically ill. A cohort of Ednrb-/- mice were observed until they developed clinical illness, at which time they were euthanized and had multiple organ homogenates cultured for bacteria, and colon and small bowel were histopathologically graded for enterocolitis. Spearman's rank correlations comparing enterocolitis scores with level of bacteremia were performed.Results: Intra-and interrater ICCs of the histologic scoring system were satisfactory (0.61 and 0.94, respectively), as were intra-and interrater CVs (18% and 9%, respectively). Of the Ednrb-/- mice, 65% developed bacteremia. Those with bacteremia had significantly higher enterocolitis scores than those without bacteremia (P < .01). Ednrb-/- mice that developed bacteremia showed a strong positive correlation between total enterocolitis scores and number of bacterial colony forming units in peritoneal lavage, liver, kidney, and aerobic spleen.Conclusions: The Ednrb-/- mouse with aganglionosis develops enterocolitis and has features similar to Hirschsprung-associated enterocolitis in humans. Our grading system is a reliable way to assess enterocolitis. By performing microsurgical pull-through, we can now perform controlled, hypothesis-driven, mechanistic studies to evaluate etiologic factors affecting enterocolitis in the Ednrb-/- mouse. (C) 2010 Elsevier Inc. All rights reserved.