Ha Se Eun, Wei Lai, Jorgensen Brian G, Lee Moon Young, Park Paul J, Poudrier Sandra M, Ro Seungil
Department of Physiology and Cell Biology, University of Nevada, Reno School of Medicine.
Department of Physiology, Wonkwang Digestive Disease Research Institute and Institute of Wonkwang Medical Science, School of Medicine, Wonkwang University.
J Vis Exp. 2018 Mar 5(133):57381. doi: 10.3791/57381.
Intestinal obstructions, that impede or block peristaltic movement, can be caused by abdominal adhesions and most gastrointestinal (GI) diseases including tumorous growths. However, the cellular remodeling mechanisms involved in, and caused by, intestinal obstructions are poorly understood. Several animal models of intestinal obstructions have been developed, but the mouse model is the most cost/time effective. The mouse model uses the surgical implantation of an intestinal partial obstruction (PO) that has a high mortality rate if it is not performed correctly. In addition, mice receiving PO surgery fail to develop hypertrophy if an appropriate blockade is not used or not properly placed. Here, we describe a detailed protocol for PO surgery which produces reliable and reproducible intestinal obstructions with a very low mortality rate. This protocol utilizes a surgically placed silicone ring that surrounds the ileum which partially blocks digestive movement in the small intestine. The partial blockage makes the intestine become dilated due to the halt of digestive movement. The dilation of the intestine induces smooth muscle hypertrophy on the oral side of the ring that progressively develops over 2 weeks until it causes death. The surgical PO mouse model offers an in vivo model of hypertrophic intestinal tissue useful for studying pathological changes of intestinal cells including smooth muscle cells (SMC), interstitial cells of Cajal (ICC), PDGFRα, and neuronal cells during the development of intestinal obstruction.
肠梗阻会阻碍或阻断蠕动,其可由腹部粘连以及包括肿瘤生长在内的大多数胃肠道疾病引起。然而,人们对肠梗阻所涉及的以及由肠梗阻引起的细胞重塑机制了解甚少。已经开发了几种肠梗阻动物模型,但小鼠模型是最具成本效益/时间效益的。该小鼠模型采用手术植入肠道部分梗阻(PO),如果操作不当,死亡率很高。此外,如果不使用适当的阻断剂或放置不当,接受PO手术的小鼠不会发生肥大。在此,我们描述了一种PO手术的详细方案,该方案能产生可靠且可重复的肠梗阻,死亡率极低。该方案利用手术放置的硅胶环环绕回肠,部分阻断小肠的消化运动。部分阻塞会使肠道因消化运动停止而扩张。肠道扩张会在环的口侧诱导平滑肌肥大,这种肥大在2周内逐渐发展,直至导致死亡。手术性PO小鼠模型提供了一种肥厚性肠道组织的体内模型,可用于研究肠梗阻发展过程中肠道细胞(包括平滑肌细胞(SMC)、 Cajal间质细胞(ICC)、血小板衍生生长因子受体α(PDGFRα)和神经元细胞)的病理变化。