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评估肺炎消散情况的实验模型

Experimental Model to Evaluate Resolution of Pneumonia.

作者信息

Villabona-Rueda Andres, Wang Daniel, D'Alessio Franco R

机构信息

Division of Pulmonary Critical Care Medicine, Johns Hopkins University School of Medicine.

Engineering Management, Whiting School of Engineering, Johns Hopkins University.

出版信息

J Vis Exp. 2023 Feb 17(192). doi: 10.3791/63925.

Abstract

Acute respiratory distress syndrome (ARDS) causes acute lung injury, characterized by rapid alveolar damage and severe hypoxemia. This, in turn, leads to high morbidity and mortality. Currently, there are no pre-clinical models that recapitulate the complexity of human ARDS. However, infectious models of pneumonia (PNA) can replicate the main pathophysiological features of ARDS. Here, we describe a model of PNA induced by the intratracheal instillation of live Streptococcus pneumoniae and Klebsiella pneumoniae in C57BL6 mice. In order to evaluate and characterize the model, after inducing injury, we carried out serial measurements of body weight and bronchoalveolar lavage (BAL) for measuring markers of lung injury. Additionally, we harvested lungs for cell count and differentials, BAL protein quantification, cytospin, bacterial colony-forming unit counts, and histology. Lastly, high dimensional flow cytometry was performed. We propose this model as a tool to understand the immune landscape during the early and late resolution phases of lung injury.

摘要

急性呼吸窘迫综合征(ARDS)会导致急性肺损伤,其特征为肺泡迅速受损和严重低氧血症。这进而会导致高发病率和高死亡率。目前,尚无能够重现人类ARDS复杂性的临床前模型。然而,肺炎(PNA)感染模型可以复制ARDS的主要病理生理特征。在此,我们描述一种通过向C57BL6小鼠气管内注入活的肺炎链球菌和肺炎克雷伯菌诱导PNA的模型。为了评估和表征该模型,在诱导损伤后,我们对体重进行了连续测量,并进行支气管肺泡灌洗(BAL)以测量肺损伤标志物。此外,我们采集肺组织进行细胞计数和分类、BAL蛋白定量、细胞涂片、细菌集落形成单位计数以及组织学检查。最后,进行了高维流式细胞术检测。我们提出将此模型作为一种工具,用于了解肺损伤早期和晚期消退阶段的免疫格局。

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