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用弹性蛋白酶创建小鼠实验性腹主动脉瘤

Creation of murine experimental abdominal aortic aneurysms with elastase.

作者信息

Azuma Junya, Asagami Tomoko, Dalman Ronald, Tsao Philip S

机构信息

Department of Cardiovascular Medicine, Stanford University School of Medicine.

出版信息

J Vis Exp. 2009 Jul 23(29):1280. doi: 10.3791/1280.

Abstract

Transient intraluminal infusion of porcine pancreatic elastase into the infrarenal segment of the abdominal aorta is the most widely used animal model of abdominal aortic aneurysm (AAA) ever since it was first described in rats by Anidjar and colleagues.(1) The rationale for its development was based on the disrupted nature of elastin observed in AAAs. This rat model has been modified to produce AAAs in the infrarenal aortic region of mice.(2) The model has the ability to add broad insight into the pathobiology of AAA due to the emergence of numerous transgenic and gene knockout mice. Moreover, it is a viable platform to test potential therapeutic agents for AAA. In this video, we demonstrate the elastase infusion AAA procedure used in our laboratory. Mice are anesthetized using 2.5% isoflurane, and a laparotomy is performed under sterile conditions. The abdominal aortais isolated with the assistance of an operating stereomicroscope (Leica). After placing temporary ligatures around the proximal and distal aorta, an aortotomy is created at the bifurcation with the tip of a 30-gauge needle. A heat-tapered segment of PE-10 polyethylene tubing is introduced through the aortotomy and secured. The aortic lumen is subsequently perfused for 5-15 minutes at 100 mm Hg with saline containing type I porcine pancreatic elastase (4.5 U/mL; Sigma Chemical Co.). After removing the perfusion catheter, the aortotomy is repaired without constriction of the lumen.

摘要

自Anidjar及其同事首次在大鼠中描述以来,向腹主动脉肾下段腔内短暂注入猪胰弹性蛋白酶是腹主动脉瘤(AAA)最广泛使用的动物模型。(1)其开发的基本原理基于在AAA中观察到的弹性蛋白的破坏性质。该大鼠模型已被修改以在小鼠的肾下主动脉区域产生AAA。(2)由于出现了许多转基因和基因敲除小鼠,该模型能够为AAA的病理生物学提供广泛的见解。此外,它是测试AAA潜在治疗药物的可行平台。在本视频中,我们展示了我们实验室中使用的弹性蛋白酶注入AAA程序。使用2.5%异氟醚对小鼠进行麻醉,并在无菌条件下进行剖腹手术。在手术体视显微镜(徕卡)的辅助下分离腹主动脉。在主动脉近端和远端周围放置临时结扎线后,用30号针头的尖端在分叉处进行主动脉切开术。将一段热缩的PE-10聚乙烯管通过主动脉切开术插入并固定。随后用含有I型猪胰弹性蛋白酶(4.5 U/mL;Sigma化学公司)的盐水在100 mmHg下对主动脉腔灌注5-15分钟。取出灌注导管后,修复主动脉切开术,不使管腔狭窄。

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