Name: creation of a rodent model of abdominal aortic aneurysm by blocking adventitial vasa vasorum perfusion
Uploaded: 2017-11-08T12:30:00.000+00:00
Duration: 8 min 37 s
Description: Watch this Scientific Journal Video about Creation of a Rodent Model of Abdominal Aortic Aneurysm by Blocking Adventitial Vasa Vasorum Perfusion at JoVE.com

这项工作得到了补助金研究 (B) (20291958) 至 N.U. 的支持;青年科学家补助金 (一) (25713024) 新西兰

动物护理和实验是按照滨大学医学院动物保育委员会在动物保育中心的指导方针进行的. 
 1. 创建模型的外科程序 注意: 将手术器械放在十年代术后的念珠灭菌器中。使用无菌手套术. 
<ol> <li> 使用雄性大鼠重 300-400 克. 允许老鼠获得食物和水 ad 随意 . </li>
	<li> 麻醉吸入异氟醚的大鼠 (2.0-3.0 毫升/升)。用脚趾捏确认适当的麻醉. </li>
	<li> 用电动剃须刀刮毛, 用酒精和聚维酮碘溶液擦洗腹部. </li>
	<li> 将老鼠放在手术台上的仰卧位。用兽医软膏对大鼠和 #39 的眼睛, 以防止在麻醉时干燥. </li>
	<li> 使用剪刀在腹中线腹部切口进行剖腹探查。为了确保一个明确的外科领域, 用无菌纱布将腹腔内的内容用伤口牵引器包起来. </li>
	<li> 将主动脉从血管组织中分离出来, 用镊子轻轻拿起并撕裂后, 露出主动脉壁, 在腹膜后间隙中从左肾静脉的水平上剥落下主动脉, 从血管组织 ( 图 1A ). </li>
	<li> 结扎的血管从腹主动脉分支, 用5-0 丝线阻断血液供应, 远离主动脉, 不缩小主动脉腔. </li>
	<li> 阻断主动脉血流, 将血管夹放在肾动脉下方, 刚好位于主动脉的分岔处. 
	注: 夹的大小应大于主动脉直径, 以完全阻断血流。临时5-0 丝线结扎也足以阻断血流而不是夹子. </li>
	<li> 类似于在主动脉壁面上产生点出血, 使用微剪刀和 #160 切断前主动脉壁; 从主动脉夹远端5毫米, 毗邻肾动脉分支, 插入聚氨酯导管. </li>
	<li> 插入聚氨酯导管 (外径0.55 毫米, 内径 0.37 mm), 使用24口径留置针, 通过切口, 在主动脉上用清水冲洗掉血液。将1毫升的注射器灌入聚氨酯导管, 用水冲洗主动脉中剩余的血液。冲洗后, 从主动脉中取出导管. </li>
	<li> 在插入聚氨酯导管之前 (外径 1.20 mm, 内径 0.94 mm) 使用18口径留置针, 将聚氨酯导管切割为 10 mm ( 图 1B )。
	<ol> <li> 将聚氨酯导管的切片插入1.9 步中所做的切口的主动脉腔内。将10毫米导管完全置于主动脉腔内, 并将导管的中点重新定位, 以匹配切口位置 ( 即 , 用导管覆盖切口). </li>
	</ol> </li> <li> 用中断的 #160 修复切口; 使用8-0 单丝字符串 ( 图 1C ) 进行缝合. </li>
	<li> 结扎腹主动脉采用5-0 丝柱和聚氨酯导管的组合 ( 图 1D ). 
	注: 主动脉腔可由留置导管维持。结扎主动脉, 以防止美国导管位置。外科医生和 #39 的结是推荐的, 因为它增加了一个额外的扭曲, 当绑第一次抛出的结, 形成一个双上手结, 一个额外的回合提供更多的摩擦, 可以减少松动. </li>
	<li> 结扎后, 取出主动脉分岔处的血管夹, 然后取出肾动脉下方的血管夹, 恢复顺血流。确认总主动脉搏动. </li>
	<li> 关闭腹部切口两层, 腹膜等层, 用4-0 聚丙烯缝合。严密缝合切口以防止凸出的器官. </li>
	<li> 在腹部切口上应用局部利多卡因。把老鼠放在暖气垫上直到意识恢复。不要把老鼠单独留在那里, 直到它恢复了足够的意识来维持胸骨卧床。在完全恢复之前, 不要把已经经过手术的老鼠退回给其他老鼠. </li>
	<li> 操作后密切监视鼠。如果老鼠表现出痛苦或体重下降的迹象, 就在皮下注射不育生理盐水 (1.0-2.0 毫升). </li>
	<li> 术后, 观察时效扩张主动脉在超声图的发育。如 5 所述, 测量腹主动脉的内侧边缘到外边缘的最大直径. </li>
</ol> 2。收割、固定和橡皮-van Gieson (EVG) 染色 <ol> <li> 术后二十八天, 管理戊巴比妥钠和 #8805; 100 毫克/千克腹腔安乐鼠. </li>
	<li> 切开大鼠和 #39 的腹腔腹。用手术刀做最初的切口。用剪刀收割动脉瘤, 将收获的脉放在10% 中性缓冲的福尔马林中, 用于 24-48 小时. 
	注意: 在暴露动脉瘤时, 外科医生必须小心预防主动脉和其他器官的损伤。应观察高腹器官, 表明 AAA 形成。 图 2 显示了一个 活体 图像和 ex 体内 图像;这个代表性的科学促进会样本没有破裂. </li>
	<li> EVG 着色 9 <ol> <li> 解决方案准备 <ol> <li> 准备 Verhoeff 和 #39; s 苏木精通过在给定的顺序中混合如下: 酒精苏木素20毫升, 10% 氯化铁8.0 毫升,卢戈和 #39 的碘8毫升。在每个加法之间混合解决方案。此解决方案可以根据需要重新准备. </li>
			<li> 通过混合1% 酸品红1毫升和饱和苦味酸酸45毫升制备 Van Gieson 和 #39;允许站在一夜之间, 混合好。此解决方案稳定2周. </li>
			<li> 在40毫升蒸馏水中混合 10% FeCl 3 10 毫升, 以制备鉴别溶液 (2% 氯化铁)。此解决方案可能会根据需要重新准备. </li>
			<li> 在100毫升蒸馏水中溶解5.0 克硫代硫酸钠, 以制备5% 的亚硫酸钠。这个解决方案是稳定的1年. </li>
		</ol> </li> <li> 着色过程 注意: 有关详细信息, 请参阅参考 9 。
		<ol> <li> 简要介绍了在蒸馏水中脉的固定石蜡切片10分钟, 将脉放在 Verhoeff 和 #39 中, 用清水洗涤25分钟, 在鉴别溶液中浸泡10-30 次 (2% FeCl 3 )。在水中冲洗。在5% 的地方, 1 分钟, 在水中洗涤。在 Van Gieson 和 #39 的解决方案中放置5分钟 </li>
			<li> 在酒精中脱水, 按以下顺序: 75%、90%、95%、100% 和100% 酒精分别为三十年代。然后, 在二甲苯中清除5分钟, 两次。在片上放置树脂安装介质. 
			注: 弹力纤维呈 blue-black 至黑色;细胞核: 蓝色到血乳酸ck;胶原蛋白: 红色;和其他组织元素: 黄色. </li>
		</ol>
		</li>
	</ol>
	</li>
</ol>

<ol>
	<li>Wolinsky, H., Glagov, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+abdominal+and+thoracic+aortic+medial+structure+in+mammals.+Deviation+of+man+from+the+usual+pattern.">Comparison of abdominal and thoracic aortic medial structure in mammals. Deviation of man from the usual pattern.</a> Circ Res. 25 (6), 677-686 (1969).</li><li>Sano, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lymphangiogenesis+and+angiogenesis+in+abdominal+aortic+aneurysm.">Lymphangiogenesis and angiogenesis in abdominal aortic aneurysm.</a> PLoS One. 9 (3), e89830 (2014).</li><li>Vorp, D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Association+of+intraluminal+thrombus+in+abdominal+aortic+aneurysm+with+local+hypoxia+and+wall+weakening.">Association of intraluminal thrombus in abdominal aortic aneurysm with local hypoxia and wall weakening.</a> J Vasc Surg. 34 (2), 291-299 (2001).</li><li>Tanaka, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adventitial+vasa+vasorum+arteriosclerosis+in+abdominal+aortic+aneurysm.">Adventitial vasa vasorum arteriosclerosis in abdominal aortic aneurysm.</a> PLoS One. 8 (2), e57398 (2013).</li><li>Tanaka, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypoperfusion+of+the+Adventitial+Vasa+Vasorum+Develops+an+Abdominal+Aortic+Aneurysm.">Hypoperfusion of the Adventitial Vasa Vasorum Develops an Abdominal Aortic Aneurysm.</a> PLoS One. 10 (8), e0134386 (2015).</li><li>Tanaka, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+Mass+Spectrometry+Reveals+a+Unique+Distribution+of+Triglycerides+in+the+Abdominal+Aortic+Aneurysmal+Wall.">Imaging Mass Spectrometry Reveals a Unique Distribution of Triglycerides in the Abdominal Aortic Aneurysmal Wall.</a> J Vasc Res. 52 (2), 127-135 (2015).</li><li>Kugo, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+preventive+effect+of+fish+oil+on+abdominal+aortic+aneurysm+development.">The preventive effect of fish oil on abdominal aortic aneurysm development.</a> Biosci Biotechnol Biochem. 80 (6), 1186-1191 (2016).</li><li>Kugo, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adipocyte+in+vascular+wall+can+induce+the+rupture+of+abdominal+aortic+aneurysm.">Adipocyte in vascular wall can induce the rupture of abdominal aortic aneurysm.</a> Sci Rep. 6, 31268 (2016).</li><li>Lichtenberg, A., Cebotari, S., Tudorache, I., Hilfiker, A., Haverich, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+scaffolds+for+heart+valve+tissue+engineering.">Biological scaffolds for heart valve tissue engineering.</a> Methods Mol Med. 140, 309-317 (2007).</li><li>Heistad, D. 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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Creation+of+murine+experimental+abdominal+aortic+aneurysms+with+elastase.">Creation of murine experimental abdominal aortic aneurysms with elastase.</a> J Vis Exp. (29), (2009).</li><li>Lu, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subcutaneous+Angiotensin+II+Infusion+using+Osmotic+Pumps+Induces+Aortic+Aneurysms+in+Mice.">Subcutaneous Angiotensin II Infusion using Osmotic Pumps Induces Aortic Aneurysms in Mice.</a> J Vis Exp. (103), (2015).</li><li>United Kingdom, E. T. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endovascular+versus+open+repair+of+abdominal+aortic+aneurysm.">Endovascular versus open repair of abdominal aortic aneurysm.</a> N Engl J Med. 362 (20), 1863-1871 (2010).</li></ol>

作者没有什么可透露的。

在生理条件下, 主动脉壁的内层通过从腔血流中扩散滋养, 而外层和中层则由膜滋养, 由其渗透到内侧的1。腹主动脉壁的血流可来源于以下三方向/区域: (1) 通过主动脉壁的近端方向, (2) 通过主动脉壁的远端方向, (3) 血管组织10。以前, 我们对人体组织的组织学分析发现, 在 AAA 壁上有明显的狭窄或闭塞, 这表明腹主动脉壁的血流量可以减少4。这是一个非常重要的一点, 在这个协议, 一个下 AAA 是由聚氨酯导管插入和缝合结扎的下腹主动脉的组合造成的。为了仔细去除组织层, 外科医生必须顺利地插入一个聚氨酯导管进入主动脉和坚定地结扎主动脉, 造成慢性缺氧, 由于灌注的外和动脉瘤的形成。利用这些技术, 主动脉壁的血流减少, 诱发局部缺氧环境。血流减少和缺氧诱发的动脉瘤形成表明腹主动脉壁的血流在 AAA 形成的发病机制中起着重要作用。
具体而言, 主动脉瘤模型必须满足以下条件: 与基线相比, 血管直径增加1.5 倍, 并与基质的变性和主动脉壁的炎症有关。最流行的动物模型已经建立了诱导炎症反应使用的物质, 如 CaCl211, 弹性蛋白酶12, 和血管紧张素 II13。这些模型具有很高的重现性, 明显引起病理变化, 在研究中得到了广泛的应用。在我们的模型, 我们评估的主动脉直径使用超声检查每7天之前的程序是执行前28天的过程 (图 3)。结果表明, 在28天内, 主动脉直径适度增加, 这表明直径的变化与以前的啮齿动物模型相似。血管形态的大体观察表明, 纺锤形形状平滑 (图 2)。在28天, 我们牺牲了老鼠, 并进行了病理学分析的主动脉组织恢复。观察了粘膜和膜的弹性和胶原纤维的撕裂和消失 (图 4)。此外, 炎症细胞, 如巨噬细胞膜, 从膜到膜介质。
目前, 为科学促进会提供的治疗方案仅限于外科修复和血管内支架移植, 死亡率为 30-50% 的患者的 AAA 破裂14。然而, 没有药物被批准用于临床治疗科学促进会。有人争论说, 在 AAA 研究中, 人类与已建立的动物模型之间的病理结果存在差异。人类 aaa 和动物 aaa 模型发病机制的相似性是药理治疗发展的关键。关于啮齿类动物模型的有效性, 我们的大鼠模型在形态上与人相似, 从腔内血栓5和成8。此外, 这项研究中约有20% 的大鼠在手术后28天内就会破裂并死亡。虽然主动脉瘤破裂是这一疾病的最关键事件, 但破裂是罕见的建立实验 AAA 模型, 并尚未阐明的机制。因此, 该模型有助于了解动脉内径扩张和动脉瘤破裂的机理。
这个模型的创建是需要一些外科手术。因此, 研究人员必须练习创建此模型, 这是该模型的一个局限性。在未来, 我们希望建立一个啮齿动物模型, 我们可以通过逐渐加厚血管壁来减少血流量, 导致自发性主动脉瘤。

腹主动脉由以下三层组成: 内血管壁 (内中膜)、内侧层 (介质) 和外血管壁 (膜), 膜有独特的供血系统, 称为瓦萨血管。主动脉组织提供氧气通过外血流灌注和简单的氧扩散从主动脉血液流量1。然而, 在地理上, 腹主动脉的分布 VVs 与其他部位的主动脉相比是最少的。2
先前的研究报告的组织缺氧的人腹主动脉瘤 (AAA) 壁厚腔内血栓 (ILT)3。此外, 它已经表明, 在动脉瘤壁的外的一个人闭塞与动脉硬化的变化率明显较高, 这是与组织缺氧的 AAA 墙壁4。基于这些发现, 通过诱导外灌注5, 建立了一种新的啮齿动物模型。在这个模型中, 灌注导致组织缺氧和下 aaa 的发展, 它的形态学和病理特征与人类 aaa 级6相似。主要例子是存在的协会和增生性脂肪细胞的积累6, 并可能导致破裂7,8。这些发现在以前的啮齿动物模型中很少见到。因此, 此模型可能会大大有助于加深对负责 AAA 发展和破裂的机制的理解。我们介绍和描述一个标准的协议, 用于诱导科学促进会通过外灌注, 我们解释如何诱导缺氧的主动脉壁使用外科技术。

<table><tbody><tr><td>rat</td><td>Japan SLC.Inc</td><td>Slc:SD rat</td><td>Sprague&amp;ndash;Dawley ratTM</td></tr><tr><td>povidone-iodine solution</td><td>Libatape Pharmaceutical Co., Ltd.</td><td>4987335 111457</td><td></td></tr><tr><td>5-0 silk string</td><td>Akiyama Medical MFG. CO.,LTD</td><td>JIS No.1</td><td></td></tr><tr><td>vascular clips</td><td>Natsume Seisakusho Co., Ltd.</td><td>C-42-S-2</td><td></td></tr><tr><td>polyurethane catheter (24-gauge indwelling needle)</td><td>MEDIKIT</td><td>24G</td><td>Supercath Z4VTM, 24-gauge indwelling needle</td></tr><tr><td>polyurethane catheter (18-gauge indwelling needle)</td><td>MEDIKIT</td><td>18G</td><td>Supercath Z3VTM, 18-gauge indwelling needle</td></tr><tr><td>8-0 monofilament string</td><td>Ethicon Suture</td><td>c-42-S-2</td><td>PROLENE Polypropylene Suture, Repair the incision with the suture</td></tr></tbody></table>

creation of a rodent model of abdominal aortic aneurysm by blocking adventitial vasa vasorum perfusion

外瓦萨血管为主动脉壁提供氧气和营养。主动脉壁缺氧可引起腹主动脉瘤的扩大。本文介绍和描述了一个标准的协议, 诱导通过外灌注创建的联合聚氨酯导管插入到主动脉腔和缝合结扎下腹主动脉。
该议定书涉及使用的雄性大鼠体重300-400 克, 这是提供食物和水的ad 随意。腹腹中线切开术后, 主动脉剥离, 阻断血管组织的血流。Aortotomy 涉及一个小切口毗邻肾动脉分支执行, 和聚氨酯导管插入使用18口径留置针。在修复切口后, 在导管上结扎主动脉, 通过主动脉壁将血流从近端方向穿过, 而不会干扰主动脉血流。这种技术可以诱发主动脉扩张的 AAA 级。
这个模型最大的好处是, 灌注导致组织缺氧和发展的下 aaa, 它的形态学和病理特征类似于人类 aaa。

所描述的手术技术建立了一个新的动物模型的慢性主动脉缺氧诱发动脉瘤的结合聚氨酯导管插入和缝合结扎下腹主动脉在大鼠。在协议部分中描述的老鼠在手术后28天被安乐死。脉被采集并成像以可视化动脉瘤形成。图 2显示了梭形 AAA 的发展。在前体中, 主动脉的上、下端有正常直径而不扩张。用经腹超声 (图 3) 测量主动脉直径。直径通常达到它的最大大小在大约14天在做法以后;此后, 它保持不变或略有增加。图 4显示了动脉瘤在 EVG 染色后最大直径的组织病理图像。28天 (动脉瘤) 组织的图像显示, 与0天相比, 弹力纤维明显退化。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55763/55763fig1.jpg"> 
图 1: 诱发腹主动脉瘤 (AAA) 的手术方法。 (A) 下主动脉从周围组织脱落。(B) 一根10毫米长的聚氨酯导管通过小切口插入主动脉。(C) 切口用8-0 单丝缝线修复, 血流恢复。(D) 主动脉在插入导管上结扎5-0 丝缝线。缩放条 = 5 mm<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55763/55763fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55763/55763fig2.jpg"> 
图 2: 术后代表性结果。 术后28天的宏观观察显示梭形腹主动脉瘤的发展。后的升高的边缘对应于动脉瘤的外缘 (残破的线; 左)。在ex 体内中, 主动脉的上、下端是正常的 (右)。缩放条 = 3 mm<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55763/55763fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/55763/55763fig3.jpg"> 
图 3: 经腹超声测量的最大主动脉直径。 在大鼠模型中, 主动脉直径逐渐增加。主动脉直径为平均±标准差 (n = 12)。<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55763/55763fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/55763/55763fig4.jpg"> 
图 4: 橡皮 van Gieson 染色的动脉瘤组织的代表性图像。 EVG 染色显示退行性弹性椎板在培养基中的组织学评价及腔内血栓形成28天后的程序 (右)。28天, 主动脉膜中的弹力纤维碎裂和稀疏胶原纤维被观察到。0天是在过程之前 (左)。缩放栏 = 500 µm.<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55763/55763fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Creation of a Rodent Model of Abdominal Aortic Aneurysm by Blocking Adventitial Vasa Vasorum Perfusion at JoVE.com

Creation of a Rodent Model of Abdominal Aortic Aneurysm by Blocking Adventitial Vasa Vasorum Perfusion

聚氨酯导管插入主动脉腔, 结扎主动脉诱发慢性缺氧, 灌注外瓦萨血管。本文介绍了一种新的动物模型腹主动脉瘤 (aaa) 的特点类似的 aaa 在人。

阻断外瓦萨血管灌注对腹主动脉瘤鼠模型的建立

The adventitial vasa vasorum (VV) provides oxygen and nourishment to the aortic wall. Hypoxia in the aortic wall can cause enlarged abdominal aortic ...

creation-rodent-model-abdominal-aortic-aneurysm-blocking-adventitial

Universidad San Sebastian

Research

JoVE Journal

Medicine

9.0K 次观看.  Hamamatsu University School of Medicine. 聚氨酯导管插入主动脉腔, 结扎主动脉诱发慢性缺氧, 灌注外瓦萨血管。本文介绍了一种新的动物模型腹主动脉瘤 (aaa) 的特点类似的 aaa 在人。

视频: Creation of a Rodent Model of Abdominal Aortic Aneurysm by Blocking Adventitial Vasa Vasorum Perfusion

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

Creation of Murine Experimental Abdominal Aortic Aneurysms with Elastase 

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.

Creation of Murine Experimental Abdominal Aortic Aneurysms with Elastase

This video shows how to induce abdominal aortic aneurysms (AAA) in mice via transient intraluminal infusion of porcine pancreatic elastase into the infrarenal segment of the abdominal aorta. 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.

创建小鼠实验与弹性蛋白酶的腹主动脉瘤

Transient intraluminal infusion of porcine pancreatic elastase into the infrarenal segment of the abdominal aorta is the most widely used animal model of ...

生物学

A New Murine Model of Endovascular Aortic Aneurysm Repair

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of Abdominal aortic aneurysm (AAA) pathophysiology. There is no abdominal aortic aneurysm endograft exclusion model cheap and reproducible, which would allow deep investigations of AAA before and after treatment. We hereby describe how to induce, and then to exclude with a covered coronary stentgraft an abdominal aortic aneurysm in a rat. The well known elastase induced AAA model was first reported in 19901 in a rat, then described in mice2. Elastin degradation leads to dilation of the aorta with inflammatory infiltration of the abdominal wall and intra luminal thrombus, matching with human AAA. Endovascular exclusion with small covered stentgraft is then performed, excluding any interactions between circulating blood and the aneurysm thrombus. Appropriate exclusion and stentgraft patency is confirmed before euthanasia by an angiography thought the left carotid artery. Partial control of elastase diffusion makes aneurysm shape different for each animal. It is difficult to create an aneurysm, which will allow an appropriate length of aorta below the aneurysm for an easy stentgraft introduction, and with adequate proximal and distal neck to prevent endoleaks. Lots of failure can result to stentgraft introduction which sometimes lead to aorta tear with pain and troubles to stitch it, and endothelial damage with post op aorta thrombosis. Giving aspirin to rats before stentgraft implantation decreases failure rate without major hemorrhage. Clamping time activates neutrophils, endothelium and platelets, and may interfere with biological analysis.

Histological and biochemical modifications after aneurysm endograft exclusion are still unclear. We describe a new model of endograft implantation on a murine aneurysm. Through thrombus analysis with and without persistant circulating blood stream interface, and new endovascular biomaterials evaluation, this model will help to better understand endovascular aneurysm exclusion pathobiology.

一种新的小鼠模型主动脉瘤腔内修复

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of ...

The Helsinki Rat Microsurgical Sidewall Aneurysm Model

Experimental saccular aneurysm models are necessary for testing novel surgical and endovascular treatment options and devices before they are introduced into clinical practice. Furthermore, experimental models are needed to elucidate the complex aneurysm biology leading to rupture of saccular aneurysms.
Several different kinds of experimental models for saccular aneurysms have been established in different species. Many of them, however, require special skills, expensive equipment, or special environments, which limits their widespread use. A simple, robust, and inexpensive experimental model is needed as a standardized tool that can be used in a standardized manner in various institutions.
The microsurgical rat abdominal aortic sidewall aneurysm model combines the possibility to study both novel endovascular treatment strategies and the molecular basis of aneurysm biology in a standardized and inexpensive manner. Standardized grafts by means of shape, size, and geometry are harvested from a donor rat&#39;s descending thoracic aorta and then transplanted to a syngenic recipient rat. The aneurysms are sutured end-to-side with continuous or interrupted 9-0 nylon sutures to the infrarenal abdominal aorta.
We present step-by-step procedural instructions, information on necessary equipment, and discuss important anatomical and surgical details for successful microsurgical creation of an abdominal aortic sidewall aneurysm in the rat.

Microsurgical sidewall aneurysms in rats are created by end-to-side anastomosis of an aortic graft to the abdominal aorta. We present step-by-step instructions and discuss anatomical and surgical details for successful experimental saccular aneurysm creation.

赫尔辛基鼠显微旁瘤模型

Experimental saccular aneurysm models are necessary for testing novel surgical and endovascular treatment options and devices before they are introduced ...

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of aortic disease exist, in vivo imaging of the vasculature has been limited. In recent years, micro-computerized tomography (micro-CT) has emerged as a preferred modality for imaging both large and small vessels both in vivo and ex vivo. In conjunction with a method of vascular casting, we have successfully used micro-CT to characterize the frequency and distribution of aortic pathology in &#946;-aminopropionitrile-treated C57/Bl6 mice. Technical limitations of this method include variations in the quality of the perfusion introduced by poor animal preparation, the application of proper methodologies for vessel size quantification, and the non-survivability of this procedure. This article details a methodology for the intravascular perfusion of a lead-based radiopaque silicone rubber for the quantitative characterization of aortopathy in a mouse model of aneurysm and dissection. In addition to visualizing aortic pathology, this method may be used for examining other vascular beds in vivo or vascular beds removed post-mortem.

This article describes a detailed methodology of using a radiopaque lead-based silicone rubber to perfuse the murine vasculature for aortic diameter quantification in a mouse model of aortic aneurysm and dissection.

β-aminopropionitrile 诱发主动脉瘤和解剖小鼠模型 Aortopathy 的定量显微 CT 分析

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. However, only minor improvement in the long-term results of transplanted solid organs could be observed over the past decades. In this context, chronic allograft vasculopathy (CAV) still represents the leading cause of late organ failure in cardiac, renal and pulmonary transplantation.
Thus far, the underlying pathogenesis of CAV development remains unclear, explaining why effective treatment strategies are presently missing and emphasizing a need for relevant experimental models in order to study the underlying pathophysiology leading to CAV formation. The following protocol describes a murine heterotopic cervical aortic transplantation model using a modified non-suture cuff technique. In this technique, a segment of the thoracic aorta is interpositioned in the right common carotid artery. With the use of the non-suture cuff technique, an easy to learn and reproducible model can be established, minimizing the possible heterogeneity of sutured vascular micro anastomoses.

Here, we present a protocol of heterotopic aortic transplantation in mice using the non-suture cuff technique in a cervical murine model. This model can be used to study the underlying pathology of chronic allograft vasculopathy (CAV) and can help evaluate new therapeutic agents in order to prevent its formation.

使用改性非缝合铐技术的宫颈主动脉移植模型

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...

Porcine Model of Infrarenal Abdominal Aortic Aneurysm

Large animal models to study abdominal aortic aneurysms are sparse. The purpose of this model is to create reproducible, clinically significant infrarenal abdominal aortic aneurysms (AAA) in swine. To achieve this, we use a combination of balloon angioplasty, elastase and collagenase, and a lysyl oxidase inhibitor, called &#946;-aminopropionitrile (BAPN), to create clinically significant infrarenal aortic aneurysms, analogous to human disease.
Noncastrated male swine are fed BAPN for 7 days prior to surgery to achieve a steady state in the blood. A midline laparotomy is performed and the infrarenal aorta is circumferentially dissected. An initial measurement is recorded prior to aneurysm induction with a combination of balloon angioplasty, elastase (500 units)/collagenase (8000 units) perfusion, and topical elastase application. Swine are fed BAPN daily until terminal procedure on either postoperative day 7, 14, or 28, at which time the aneurysm is measured, and tissue procured. BAPN + surgery pigs are compared to pigs that underwent surgery alone.
Swine treated with BAPN and surgery had a mean aortic dilation of 89.9% &#177; 47.4% at day 7, 105.4% &#177; 58.1% at day 14, and 113.5% &#177; 30.2% at day 28. Pigs treated with surgery alone had significantly smaller aneurysms compared to BAPN + surgery animals at day 28 (p &#60; 0.0003). The BAPN + surgery group had macroscopic and immunohistochemical evidence of end stage aneurysmal disease.
Clinically significant infrarenal AAA can be induced using balloon angioplasty, elastase/collagenase perfusion and topical application, supplemented with oral BAPN. This model creates large, clinically significant AAA with hallmarks of human disease. This has important implications for the elucidation of AAA pathogenesis and testing of novel therapies and devices for the treatment of AAA. Limitations of the model include variation in BAPN ingested by swine, quality of elastase perfusion, and cost of BAPN.&#160;

This novel model creates robust infrarenal abdominal aortic aneurysms in swine using a combination of balloon angioplasty, elastase/collagenase perfusion, topical elastase application, and oral compound &#946;-aminopropionitrile administration, which interferes with collagen cross-linking.

肾上腺腹动脉瘤的猪模型

Large animal models to study abdominal aortic aneurysms are sparse. The purpose of this model is to create reproducible, clinically significant infrarenal ...

<meta charset="utf8"></head><body>研究小鼠腹主动脉瘤的分子机制

A Mouse Abdominal Aortic Aneurysm Model by Periadventitial Calcium Chloride and Elastase Infiltration

Abdominal aortic aneurysm (AAA) is a life-threatening disease associated with high mortality rates. It is characterized by the permanent dilation of the abdominal aorta with at least a 50% increase in arterial diameter. Various animal models of AAA have been introduced to mimic the pathophysiological changes and study the underlying mechanisms of AAA. Among these models, the calcium chloride (CaCl2)- and elastase-induced AAA models are commonly used in mice. However, these methods have certain limitations. Traditional intraluminal porcine pancreatic elastase (PPE) perfusion is associated with high technical difficulty and a high rupture rate, while periadventitial administration of PPE yields inconsistent results. In addition, the CaCl2-induced AAA model lacks human AAA features, such as atherothrombosis and aneurysm rupture. Therefore, the combined application of CaCl2 and PPE has been proposed as an approach to enhance success rates and induce greater diameter increases in AAA animal models. This manuscript presents a comprehensive protocol for establishing a mouse AAA model through periaortic infiltration of PPE and CaCl2 in the infrarenal segment of the abdominal aorta. By following this protocol, we can achieve an AAA formation rate of approximately 90% with technical simplicity and reproducibility. Further ultrasound and histological experiments confirm that this model effectively replicates the morphological and pathological changes observed in human AAA.

<meta charset="utf8"></head><body>作者聚焦：腹主动脉瘤小鼠模型的开发和表征

This manuscript presents a protocol for establishing a mouse abdominal aortic aneurysm model using calcium chloride and elastase, combining the advantages of previous modeling methods. This model can be utilized to investigate the pathophysiological mechanisms underlying abdominal aortic aneurysms.

通过外膜周围氯化钙和弹性蛋白酶浸润的小鼠腹主动脉瘤模型

Abdominal aortic aneurysm (AAA) is a life-threatening disease associated with high mortality rates. It is characterized by the permanent dilation of the ...

阻断外瓦萨血管灌注对腹主动脉瘤鼠模型的建立

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此视频中的章节

主动脉原位移植大鼠模型，研究慢性血管病变的发展

创建小鼠实验与弹性蛋白酶的腹主动脉瘤

一种新的小鼠模型主动脉瘤腔内修复

赫尔辛基鼠显微旁瘤模型

β-aminopropionitrile 诱发主动脉瘤和解剖小鼠模型 Aortopathy 的定量显微 CT 分析

使用改性非缝合铐技术的宫颈主动脉移植模型

肾上腺腹动脉瘤的猪模型

通过外膜周围氯化钙和弹性蛋白酶浸润的小鼠腹主动脉瘤模型

通过外膜周围氯化钙和弹性蛋白酶浸润的小鼠腹主动脉瘤模型

通过外膜周围氯化钙和弹性蛋白酶浸润的小鼠腹主动脉瘤模型