Name: mouse abdominal aortic aneurysm model induced by perivascular application of elastase
Uploaded: 2022-02-11T15:00:00
Description: Watch this Scientific Journal Video about Mouse Abdominal Aortic Aneurysm Model Induced by Perivascular Application of Elastase at JoVE.com

我们感谢密歇根大学实验动物医学部在动物饲养和繁殖方面的帮助。本研究由NIH RO1 HL138139，NIH RO1 HL153710至J. Zhang，NIH RO1 HL109946，RO1 HL134569至Y.E. Chen，以及美国心脏协会资助20POST35110064至G. Zhao。

动物协议已获得密歇根大学机构动物护理和使用委员会（PRO00010092）的批准。雄性和雌性C57BL / 6J野生型（WT）小鼠，〜7周龄，用于实验。
1. 动物准备
<ol><li>在手术前后用标准的食物喂养小鼠（参见 材料表）。 注意：可以使用不同的菌株和年龄的小鼠。然而，建议年龄在 5.5-12 周龄范围内，以达到最大发病率。</li>
	<li>对于每只小鼠，在麻醉诱导?...

<ol>
	<li>Kent, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+practice:+Abdominal+aortic+aneurysms.">Clinical practice: Abdominal aortic aneurysms.</a> The New England Journal of Medicine. 371 (22), 2101-2108 (2014).</li><li>Karthikesalingam, 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=Mortality+from+ruptured+abdominal+aortic+aneurysms:+Clinical+lessons+from+a+comparison+of+outcomes+in+England+and+the+USA.">Mortality from ruptured abdominal aortic aneurysms: Clinical lessons from a comparison of outcomes in England and the USA.</a> Lancet. 383 (9921), 963-969 (2014).</li><li>Noel, A. 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=Ruptured+abdominal+aortic+aneurysms:+The+excessive+mortality+rate+of+conventional+repair.">Ruptured abdominal aortic aneurysms: The excessive mortality rate of conventional repair.</a> Journal of Vascular Surgery. 34 (1), 41-46 (2001).</li><li>Lederle, F. 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=Rupture+rate+of+large+abdominal+aortic+aneurysms+in+patients+refusing+or+unfit+for+elective+repair.">Rupture rate of large abdominal aortic aneurysms in patients refusing or unfit for elective repair.</a> JAMA. 287 (22), 2968-2972 (2002).</li><li>Kochanek, K. D., Xu, J., Murphy, S. L., Minino, A. M., Kung, H. 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S., Golledge, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+abdominal+aortic+aneurysm+and+their+role+in+furthering+management+of+human+disease.">Animal models of abdominal aortic aneurysm and their role in furthering management of human disease.</a> Cardiovascular Pathology. 20 (2), 114-123 (2011).</li><li>Patelis, N., 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=Animal+models+in+the+research+of+abdominal+aortic+aneurysms+development.">Animal models in the research of abdominal aortic aneurysms development.</a> Physiological Research. 66 (6), 899-915 (2017).</li><li>Senemaud, J., 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=Translational+relevance+and+recent+advances+of+animal+models+of+abdominal+aortic+aneurysm.">Translational relevance and recent advances of animal models of abdominal aortic aneurysm.</a> Arteriosclerosis, Thrombosis, and Vascular Biology. 37 (3), 401-410 (2017).</li><li>Lysgaard Poulsen, J., Stubbe, J., Lindholt, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+used+to+explore+abdominal+aortic+aneurysms:+A+systematic+review.">Animal models used to explore abdominal aortic aneurysms: A systematic review.</a> European Journal of Vascular and Endovascular Surgery. 52 (4), 487-499 (2016).</li><li>Tsui, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+models+of+abdominal+aortic+aneurysms.">Experimental models of abdominal aortic aneurysms.</a> Open Cardiovascular Medicine Journal. 4, 221-230 (2010).</li><li>Manning, M. W., Cassi, L. A., Huang, J., Szilvassy, S. J., Daugherty, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abdominal+aortic+aneurysms:+Fresh+insights+from+a+novel+animal+model+of+the+disease.">Abdominal aortic aneurysms: Fresh insights from a novel animal model of the disease.</a> Vascular Medicine. 7 (1), 45-54 (2002).</li><li>Chiou, A. C., Chiu, B., Pearce, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+aortic+aneurysm+produced+by+periarterial+application+of+calcium+chloride.">Murine aortic aneurysm produced by periarterial application of calcium chloride.</a> Journal of Surgical Research. 99 (2), 371-376 (2001).</li><li>Allaire, E., Guettier, C., Bruneval, P., Plissonnier, D., Michel, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell-free+arterial+grafts:+Morphologic+characteristics+of+aortic+isografts,+allografts,+and+xenografts+in+rats.">Cell-free arterial grafts: Morphologic characteristics of aortic isografts, allografts, and xenografts in rats.</a> Journal of Vascular Surgery. 19 (3), 446-456 (1994).</li><li>Anidjar, S., 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=Elastase-induced+experimental+aneurysms+in+rats.">Elastase-induced experimental aneurysms in rats.</a> Circulation. 82 (3), 973-981 (1990).</li><li>Sun, J., 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=Mast+cells+modulate+the+pathogenesis+of+elastase-induced+abdominal+aortic+aneurysms+in+mice.">Mast cells modulate the pathogenesis of elastase-induced abdominal aortic aneurysms in mice.</a> Journal of Clinical Investigation. 117 (11), 3359-3368 (2007).</li><li>Bhamidipati, C. 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=Development+of+a+novel+murine+model+of+aortic+aneurysms+using+peri-adventitial+elastase.">Development of a novel murine model of aortic aneurysms using peri-adventitial elastase.</a> Surgery. 152 (2), 238-246 (2012).</li><li>Pyo, R., 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=Targeted+gene+disruption+of+matrix+metalloproteinase-9+(gelatinase+B)+suppresses+development+of+experimental+abdominal+aortic+aneurysms.">Targeted gene disruption of matrix metalloproteinase-9 (gelatinase B) suppresses development of experimental abdominal aortic aneurysms.</a> Journal of Clinical Investigation. 105 (11), 1641-1649 (2000).</li><li>Tanaka, A., Hasegawa, T., Chen, Z., Okita, Y., Okada, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+rat+model+of+abdominal+aortic+aneurysm+using+a+combination+of+intraluminal+elastase+infusion+and+extraluminal+calcium+chloride+exposure.">A novel rat model of abdominal aortic aneurysm using a combination of intraluminal elastase infusion and extraluminal calcium chloride exposure.</a> Journal of Vascular Surgery. 50 (6), 1423-1432 (2009).</li><li>Busch, 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=Four+surgical+modifications+to+the+classic+elastase+perfusion+aneurysm+model+enable+haemodynamic+alterations+and+extended+elastase+perfusion.">Four surgical modifications to the classic elastase perfusion aneurysm model enable haemodynamic alterations and extended elastase perfusion.</a> European Journal of Vascular and Endovascular Surgery. 56 (1), 102-109 (2018).</li><li>Liang, W., 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=KLF11+protects+against+venous+thrombosis+via+suppressing+tissue+factor+expression.">KLF11 protects against venous thrombosis via suppressing tissue factor expression.</a> Thrombosis and Haemostasis. , (2021).</li><li>Marinov, G. 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弹性蛋白酶诱导的AAA模型首先由Anidjar等人于1990年使用大鼠报道17。在过去的三十年中，已经引入了各种改进版本，同时在手术技术方面有显着改进19，20，21，22。数百个研究所使用弹性蛋白酶诱导的AAA模型作为AAA研究的第二常用啮齿动物实验模型12。很自然，不同?...

腹主动脉瘤 （AAA） 定义为腹主动脉节段性扩张，血管直径至少增加 50%1。AAA可能是致命的，因为破裂可导致极高的死亡率，即使有干预2，3，4。据报道，AAA每年在美国造成约13，000人死亡，这使其成为第 10大死因1，5。
AAA的发病机制尚未完全了解6，7，8。为了研究AAA的分子机制并测试潜在的治疗靶点，已经建立了几个实验AAA模型9，10。AAA的啮齿动物模型包括弹性蛋白酶、氯化钙、血管紧张素II和异种移植物模型，其中弹性蛋白酶诱导的AAA模型是第二常用的模型10、11、12、13、14、15、16、17。该模型涉及直接输注或应用猪胰腺弹性蛋白酶（PPE）到主动脉的红外线段。弹性蛋白酶渗透到主动脉的内侧层将导致弹性薄片的降解和炎性细胞的浸润，导致主动脉壁完整性的丧失和随后腹主动脉7，18的扩张。弹性蛋白酶诱导的AAA模型于1990年由Anidjar等人使用大鼠首次报道，其中分离的主动脉部分灌注弹性蛋白酶17。2012年晚些时候，Bhamidipati等人报告了使用个人防护装备的腹膜应用的改进模型。如今，弹性蛋白酶诱导的AAA模型的大多数手术都受到Bhamidipati组的启发，并且通过外部应用而不是PPE的腔内灌注进行。虽然外用对精细手术技能的要求较低，但AAA的发生率相对较低，且尺寸略小于腔内灌注11，19。
虽然在AAA研究中广泛使用，但弹性蛋白酶诱导的AAA模型具有一定的局限性。该模型的一个警告是手术方式的不可避免的变化，这可能导致可重复性的问题。例如，在如何分离主动脉的红外线段以及选择该段的哪个部分用于不同实验室的PPE应用方面，外科手术中可能存在差异。PPE的酶活性和PPE孵育的持续时间也可能不同。然而，这些都是影响最终AAA形成率和动脉瘤直径的基本决定因素。这些关键决定因素的变化使得使用该模型的不同组的AAA研究的数据比较非常困难。因此，需要标准化的外科手术程序作为从各种机构获得可比结果的工具。
本文描述了弹性蛋白酶诱导的AAA模型的标准化手术方案，方法是将PPE直接应用于小鼠红外腹主动脉的外膜。还将讨论有关使用此模型在小鼠中成功和稳健地生成AAA所必需的手术材料和程序的详细信息。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>6-0 non-absorbable monofilament suture</td>
			<td>Pro Advantage</td>
			<td>P420697</td>
			<td></td>
		</tr>
		<tr>
			<td>Carprofen</td>
			<td>Zoetis Inc.</td>
			<td>NDC: 54771-8507</td>
			<td></td>
		</tr>
		<tr>
			<td>Chow Diet</td>
			<td>LabDiet</td>
			<td>3005659-220</td>
			<td>PicoLab 5L0D</td>
		</tr>
		<tr>
			<td>Cotton Applicator</td>
			<td>Dynarex</td>
			<td>4303</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton Pad</td>
			<td>Rael</td>
			<td>UPC: 810027130969</td>
			<td></td>
		</tr>
		<tr>
			<td>GraphPad Prism 8</td>
			<td>GraphPad Software Inc.</td>
			<td></td>
			<td>Version 8.4.3</td>
		</tr>
		<tr>
			<td>Grarfe Forceps</td>
			<td>Fine Science Tools</td>
			<td>11051-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Halsted Mosquito Hemostats</td>
			<td>Fine Science Tools</td>
			<td>13009-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Par Pharmaceutical</td>
			<td>NDC: 42023-0115-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrile gloves</td>
			<td>Fisherbrand</td>
			<td>19-130-1597</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin</td>
			<td>Thermo Fisher</td>
			<td>15140122</td>
			<td></td>
		</tr>
		<tr>
			<td>Porcine pancreatic elastase</td>
			<td>Sigma-Aldrich</td>
			<td>E1250-100MG</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Fine Science Tools</td>
			<td>14068-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile 0.9% saline solution</td>
			<td>Baxter</td>
			<td>2B1324X</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Akorn</td>
			<td>NDC: 59399-110-20</td>
			<td></td>
		</tr>
	</tbody>
</table>

mouse abdominal aortic aneurysm model induced by perivascular application of elastase

腹主动脉瘤 （AAA） 虽然主要无症状，但可能危及生命，因为 AAA 破裂通常具有毁灭性后果。目前，AAA有几种不同的实验模型，每种模型都强调AAA发病机制的不同方面。弹性蛋白酶诱导的AAA模型是第二常用的啮齿动物AAA模型。该模型涉及直接输注或应用猪胰腺弹性蛋白酶（PPE）到主动脉的红外线段。由于技术挑战，现在大多数弹性蛋白酶诱导的AAA模型都是通过外部应用而不是腔内输注PPE进行的。弹性蛋白酶的浸润会导致内侧层弹性薄片的降解，导致主动脉壁完整性的丧失和随后的腹主动脉扩张。然而，弹性蛋白酶诱导的AAA模型的一个缺点是手术方式的不可避免的变化。具体而言，分离主动脉红外段的手术技术、用于主动脉包裹和PPE孵育的材料、PPE的酶活性以及PPE应用的持续时间都是影响最终AAA形成速率和动脉瘤直径的重要决定因素。值得注意的是，这些因素与AAA不同研究的差异可能导致可重复性问题。本文描述了弹性蛋白酶诱导的AAA模型的详细手术过程，方法是将PPE直接应用于小鼠红外腹主动脉的外膜。按照这一程序，雄性和雌性小鼠的稳定AAA形成率约为80%是可以实现的。通过建立标准外科手术，使用弹性蛋白酶诱导的AAA模型的AAA研究的一致性和可重复性可以显着提高。

总共有23只7周龄的野生型（WT）小鼠，包括12只雌性和11只雄性，按照所提出的方案进行手术。存活率为100%（不包括手术死亡率）。最大腹主动脉直径由卡尺测量。
AAA被定义为腹主动脉扩张，血管直径增加50%。因此，选择腹主动脉最大直径增加 50% 作为成功 AAA 诱导的临界点。根据这一标准，雌性动物术后第14天AAA的发生率为91.7%，接受手术的12只动物中有11只发展为AAA;而雄性动...

Watch this Scientific Journal Video about Mouse Abdominal Aortic Aneurysm Model Induced by Perivascular Application of Elastase at JoVE.com

Mouse Abdominal Aortic Aneurysm Model Induced by Perivascular Application of Elastase

本方案描述了一种标准化的手术方法，通过弹性蛋白酶直接应用于小鼠红外腹主动脉的外来，用于弹性蛋白酶诱导的AAA模型。

血管周围应用弹性蛋白酶诱导的小鼠腹主动脉瘤模型

Abdominal aortic aneurysm (AAA), although primarily asymptomatic, is potentially life-threatening as the rupture of AAA usually has a devastating outcome. ...

这是弹性蛋白酶诱导腹主动脉瘤的标准化方案，这将大大提高使用该模型的研究的可重复性和一致性。该技术具有一致的腹主动脉瘤诱导率。它还具有费用低，运行时间短的优点，这将产生更强的统计能力和可重复性。您需要确保去除覆盖主动脉的结缔组织，以便弹性蛋白酶有更好的渗透机会，但要注意不要损坏血管。首先，在麻醉诱导前30分钟皮下注射每公斤卡洛芬5毫克。在皮肤切口前测试麻醉的小鼠缺乏脚趾夹紧反应。沿着中下腹部的中线在皮肤上做一个2.5厘米的纵向切口。轻轻拉起下面的肌肉，沿着白线做一个2.5厘米的纵向切口。使用湿润的棉头涂抹器将肠道和胃移动到小鼠的右侧，并进入腹腔。使用镊子轻轻去除覆盖腹主动脉和下腔静脉的结缔组织。使用镊子轻轻地从下面的肌肉中解剖腹主动脉的背面和下腔静脉。将一块四厘米乘四毫米的手套条纹穿过腹主动脉和下腔静脉的背面，并拉直条纹。将条纹放置在距离右肾动脉约0.5厘米的地方。在条纹上方，通过腹主动脉和下腔静脉的背面放置一块三厘米乘两毫米的棉垫。然后，拉直化妆棉。要孵育弹性蛋白酶，使用移液管将30微升猪胰腺弹性蛋白酶滴到化妆棉上方的主动脉段上。将化妆棉和条纹缠绕在主动脉和下腔静脉周围。用无菌的0.9%生理盐水冲洗一块10厘米乘10厘米的纱布，并将其放在腹部。30分钟后，用镊子取下条纹和化妆棉。用500微升无菌0.9%盐水冲洗主动脉和腹腔。使用纱布吸收剩余的盐水。用运行6-0不可吸收的单丝缝合线重新近似肌肉层。然后，用3-4根中断的6-0条不可吸收的单丝缝合线闭合皮肤。雌性动物术后第14天腹主动脉瘤的发生率为91.7%，而雄性动物的发生率为72.7%，在发生腹主动脉瘤的19只动物中，雌性和雄性分别观察到最大腹主动脉直径增加1.7倍和1.6倍， 与非手术对照相比。显示术后第14天非手术组和手术组的主动脉代表性图像。该技术是帮助剖析三重A的病理生理学并帮助发现新的潜在治疗靶点的潜在工具。

Research

JoVE Journal

Biology

Creation of Murine Experimental Abdominal Aortic Aneurysms with Elastase 

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 ...

科研

生物学

In vivo Imaging and Therapeutic Treatments in an Orthotopic Mouse Model of Ovarian Cancer

In vivo Imaging and Therapeutic Treatments in an Orthotopic Mouse Model of Ovarian Cancer

活体成像和治疗卵巢癌的原位小鼠模型

Human cancer and response to therapy is better represented in orthotopic animal models.  This paper describes the development of an orthotopic mouse model ...

Generation of Mice Derived from Induced Pluripotent Stem Cells

诱导多能干细胞来源于小鼠的生成

The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

在造血干细胞和祖细胞的成年小鼠颅骨骨髓体内四维跟踪

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Isolation and Primary Culture of Mouse Aortic Endothelial Cells

隔离和鼠标主动脉内皮细胞原代培养

The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. The mouse is an ...

Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids

糖皮质激素粪分析：在马属动物无创监测肾上腺

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, ...

Trans-inner Cell Mass Injection of Embryonic Stem Cells Leads to Higher Chimerism Rates

胚胎干细胞的跨内细胞质量注射导致嵌合体率增高

In an effort to increase efficiency in the creation of genetically modified mice via ES Cell methodologies, we present an adaptation to the current ...

Differentiation Capacity of Human Aortic Perivascular Adipose Progenitor Cells

人主动脉周脂肪祖细胞的分化能力

Adipose tissue is a rich source of multi-potent mesenchymal stem cells (MSC) capable of differentiating into osteogenic, adipogenic, and chondrogenic ...