Vilz Tim O, Overhaus Marcus, Stoffels Burkhard, Websky Martin von, Kalff Joerg C, Wehner Sven
Department of Surgery, University of Bonn.
J Vis Exp. 2012 Sep 11(67):4086. doi: 10.3791/4086.
Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the complex cellular and molecular of intestinal pathology. Although the tunica mucosa is often the organ of interest in many inflammatory diseases, recent works demonstrated that the muscularis externa (ME) is also a highly immunocompetent organ that harbours a dense network of resident immunocytes.(1,2) These works were performed within the standardized model of intestinal manipulation (IM) that leads to inflammation of the bowel wall, mainly limited to the ME. Clinically this inflammation leads to prolonged intestinal dysmotility, known as postoperative ileus (POI) which is a frequent and unavoidable complication after abdominal surgery.(3) The inflammation is characterized by liberation of proinflammatory mediators such as IL-6(4) or IL-1β or inhibitory neurotransmitters like nitric oxide (NO).(5) Subsequently, tremendous numbers of immunocytes extravasate into the ME, dominated by polymorphonuclear neutrophils (PMN) and monocytes and finally maintain POI.(2) Lasting for days, this intestinal paralysis leads to an increased risk of aspiration, bacterial translocation and infectious complications up to sepsis and multi organ failure and causes a high economic burden.(6) In this manuscript we demonstrate the standardized model of IM and in vivo assessment of gastrointestinal transit (GIT) and colonic transit. Furthermore we demonstrate a method for separation of the ME from the tunica mucosa followed by immunological analysis, which is crucial to distinguish between the inflammatory responses in these both highly immunoactive bowel wall compartments. All analyses are easily transferable to any other research models, affecting gastrointestinal function.
胃肠道炎症是引发多种人类疾病的常见原因。动物研究模型对于探究肠道病理复杂的细胞和分子机制至关重要。尽管在许多炎症性疾病中,黏膜通常是关注的器官,但最近的研究表明,外肌层(ME)也是一个具有高度免疫活性的器官,其中驻留着密集的免疫细胞网络。(1,2)这些研究是在肠道操作(IM)的标准化模型中进行的,该模型会导致肠壁炎症,主要局限于ME。临床上,这种炎症会导致肠道运动功能长期紊乱,即术后肠梗阻(POI),这是腹部手术后常见且不可避免的并发症。(3)炎症的特征是促炎介质如IL-6(4)或IL-1β的释放,或抑制性神经递质如一氧化氮(NO)的释放。(5)随后,大量免疫细胞渗出到ME中,以多形核中性粒细胞(PMN)和单核细胞为主,最终导致POI。(2)这种肠道麻痹持续数天,会增加误吸、细菌移位和感染并发症的风险,直至发生败血症和多器官功能衰竭,并造成高昂的经济负担。(6)在本论文中,我们展示了IM的标准化模型以及胃肠道转运(GIT)和结肠转运的体内评估。此外,我们展示了一种将ME与黏膜分离并进行免疫分析的方法,这对于区分这两个高度免疫活性的肠壁隔室中的炎症反应至关重要。所有分析都很容易应用于任何其他影响胃肠道功能的研究模型。