Name: a calcium phosphate-induced mouse abdominal aortic aneurysm model
Uploaded: 2022-11-18T13:50:00
Description: Watch this Scientific Journal Video about A Calcium Phosphate-Induced Mouse Abdominal Aortic Aneurysm Model at JoVE.com

这项研究得到了中国国家自然科学基金（NSFC，81730010，91839302，81921001，31930056和91529203）和国家重点研发计划（2019YFA 0801600）的资助。

动物研究按照北京大学医学部机构动物护理与使用委员会的指导方针进行，并得到北京大学生物医学伦理委员会（LA2015142）的批准。所有用于手术的小鼠均用异氟醚（1.5%-2%）麻醉，并仔细监测麻醉以避免小鼠疼痛或不适。
1. 准备
<ol><li>切下0.3厘米宽的无粉橡胶手套和纱布。</li>
	<li>购买8-10周龄的C57BL / 6J雄性小鼠。将动物安置在空调环境中，进行12小时的明暗循...

<ol>
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T., 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=Influence+of+apolipoprotein+E,+age+and+aortic+site+on+calcium+phosphate+induced+abdominal+aortic+aneurysm+in+mice.">Influence of apolipoprotein E, age and aortic site on calcium phosphate induced abdominal aortic aneurysm in mice.</a> Atherosclerosis. 235 (1), 204-212 (2014).</li><li>Zhao, G., 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=Unspliced+xbp1+confers+VSMC+homeostasis+and+prevents+aortic+aneurysm+formation+via+foxo4+interaction.">Unspliced xbp1 confers VSMC homeostasis and prevents aortic aneurysm formation via foxo4 interaction.</a> Circulation Research. 121 (12), 1331-1345 (2017).</li><li>Jia, Y., 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=Targeting+macrophage+TFEB-14-3-3+epsilon+interface+by+naringenin+inhibits+abdominal+aortic+aneurysm.">Targeting macrophage TFEB-14-3-3 epsilon interface by naringenin inhibits abdominal aortic aneurysm.</a> Cell Discovery. 8 (1), 21 (2022).</li><li>Gage, G. J., Kipke, D. R., Shain, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+animal+perfusion+fixation+for+rodents.">Whole animal perfusion fixation for rodents.</a> Journal of Visualized Experiments. (65), e3564 (2012).</li><li>Yu, B., 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=CYLD+deubiquitinates+nicotinamide+adenine+dinucleotide+phosphate+oxidase+4+contributing+to+adventitial+remodeling.">CYLD deubiquitinates nicotinamide adenine dinucleotide phosphate oxidase 4 contributing to adventitial remodeling.</a> Arteriosclerosis, Thrombosis, and Vascular Biology. 37 (8), 1698-1709 (2017).</li><li>Altobelli, E., Rapacchietta, L., Profeta, V. F., Fagnano, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Risk+factors+for+abdominal+aortic+aneurysm+in+population-based+studies:+A+systematic+review+and+meta-analysis.">Risk factors for abdominal aortic aneurysm in population-based studies: A systematic review and meta-analysis.</a> International Journal of Environmental Research and Public Health. 15 (12), 2805 (2018).</li><li>Theivacumar, N. S., Stephenson, M. A., Mistry, H., Valenti, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diabetes+mellitus+and+aortic+aneurysm+rupture:+A+favorable+association.">Diabetes mellitus and aortic aneurysm rupture: A favorable association.</a> Vascular and Endovascular Surgery. 48 (1), 45-50 (2014).</li><li>Tanaka, T., Takei, Y., Yamanouchi, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperglycemia+suppresses+calcium+phosphate-induced+aneurysm+formation+through+inhibition+of+macrophage+activation.">Hyperglycemia suppresses calcium phosphate-induced aneurysm formation through inhibition of macrophage activation.</a> Journal of the American Heart Association. 5 (3), 003062 (2016).</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> Journal of Visualized Experiments. (103), e53191 (2015).</li><li>Urry, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neutral+sites+for+calcium+ion+binding+to+elastin+and+collagen:+A+charge+neutralization+theory+for+calcification+and+its+relationship+to+atherosclerosis.">Neutral sites for calcium ion binding to elastin and collagen: A charge neutralization theory for calcification and its relationship to atherosclerosis.</a> Proceedings of the National Academy of Sciences of the United States of America. 68 (4), 810-814 (1971).</li><li>Li, Z. 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CaPO4的手术周围应用是在小鼠中诱导AAA的可靠方法。一些研究使用了CaPO4模型，并一致报告说这是一种快速且可重复的方法来研究小鼠的AAA7，9。该模型被认为概括了人主动脉瘤的部分特征，并为AAA发病机制提供了机制见解，包括炎症和细胞外基质降解。
人类AAA的危险因素主要包括衰老、男性、吸烟、高脂血症、高?...

腹主动脉瘤 （AAA） 是一种致命的心血管疾病，其特征是腹主动脉永久性扩张，一旦破裂，死亡率很高。AAA 与衰老、吸烟、男性、高血压和高脂血症有关1。几种病理过程已被证明有助于AAA的形成，包括细胞外基质纤维蛋白水解，免疫细胞浸润和血管平滑肌细胞的丧失。目前，AAA的病理机制仍然难以捉摸，并且没有经过验证的药物治疗AAA1。由于人类主动脉样本很少，对人类AAA的研究受到限制;因此，已经建立并广泛采用几种化学修饰诱导的动物AAA模型，包括皮下血管紧张素II（AngII）输注，血管周围或腔内弹性蛋白酶孵育以及血管周围磷酸钙应用2。常用的小鼠模型是将磷酸钙（CaPO4）应用于肾下腹主动脉的外膜，具有成本效益且不需要基因修饰。
Gertz等人最初报道了将CaCl2直接应用于兔颈动脉以诱导动脉瘤变化，后来应用于小鼠的腹主动脉。该模型由Yamanouchi等人开发，通过在小鼠中使用CaPO 4晶体来加速主动脉扩张4。CaPO4浸润到小鼠主动脉中概括了在人类AAA中观察到的许多病理特征，包括深度巨噬细胞浸润，细胞外基质降解和钙沉积。人类AAA的危险因素，如高脂血症，也增加了CaPO4诱导的小鼠AAA5。与ApoE-/-或LDLR-/-小鼠中AngII灌注诱导的AAA相反，CaPO4诱导的AAA发生在肾下主动脉区域，其模拟人类AAA。目前，该方法已广泛应用于评估转基因小鼠对AAA发展的敏感性和评估药物6，7的抗AAA作用。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>CaCl2</td>
			<td>MECKLIN</td>
			<td>C805225</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Biomed</td>
			<td>SH5001-01</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>HARVEYBIO</td>
			<td>MB5051</td>
			<td></td>
		</tr>
		<tr>
			<td>Small animal ventilator</td>
			<td>RWD</td>
			<td>H1550501-012</td>
			<td></td>
		</tr>
	</tbody>
</table>

a calcium phosphate-induced mouse abdominal aortic aneurysm model

腹主动脉瘤 （AAA） 是一种危及生命的心血管疾病，在世界范围内发生，其特征是腹主动脉不可逆扩张。目前，使用了几种化学诱导的鼠AAA模型，每个模型都模拟AAA发病机制的不同方面。与血管紧张素II和弹性蛋白诱导的AAA模型相比，磷酸钙诱导的AAA模型是一种快速且具有成本效益的模型。将CaPO4 晶体应用于小鼠主动脉会导致弹性纤维降解，平滑肌细胞丢失，炎症和与主动脉扩张相关的钙沉积。本文介绍了CaPO4诱导的AAA模型的标准协议。该方案包括材料制备，将CaPO4手术应用于肾下腹主动脉的外膜，收获主动脉以可视化主动脉瘤以及小鼠的组织学分析。

施用CaPO4后14天，对C57BL/6J雄性小鼠实施安乐死，收获并清洁其主动脉。对主动脉的形态进行成像以可视化AAA形成。如图1A-B所示，CaPO4的应用导致肾下腹主动脉扩张。在组织学上，CaPO4导致弹性纤维的显着降解，如弹性蛋白断裂所示（图1C）。
<img alt="Figure 1" c...

Watch this Scientific Journal Video about A Calcium Phosphate-Induced Mouse Abdominal Aortic Aneurysm Model at JoVE.com

A Calcium Phosphate-Induced Mouse Abdominal Aortic Aneurysm Model

该协议描述了磷酸钙诱导的腹主动脉瘤（AAA）小鼠模型，以研究AAA的病理特征和分子机制。

磷酸钙诱导的小鼠腹主动脉瘤模型

An abdominal aortic aneurysm (AAA) is a life-threatening cardiovascular disease that occurs worldwide and is characterized by irreversible dilation of the ...

该方案描述了磷酸钙诱导的腹主动脉瘤三A小鼠模型，以研究腹主动脉瘤分子机制中的病理特征。与血管紧张素管和弹性蛋白酶诱导的腹主动脉瘤模型相比，磷酸钙诱导的腹主动脉瘤模型是一种快速且具有成本效益的模型。演示该程序的将是蔡泽宇，这里的博士后，我们实验室的研究员。开始切割0.3厘米宽的无粉橡胶手套和纱布。然后购买八到十周大的C57BL6J Mayo小鼠，并将动物饲养在空调环境中，12小时的光照黑暗循环，并免费获得食物和水。接下来，在手术前高压灭菌纱布、棉签、剪刀和镊子，并获得 Betadine、70% 乙醇和消毒洗手液。使用电动剪或脱毛膏剃除麻醉小鼠的腹毛。然后用Betadine擦拭并擦拭剃光的区域，然后以圆周运动数次70%乙醇。然后用剪刀沿着腹部中线在下腹部做一个大约 1.5 厘米的切口。使用蘸有生理盐水的无菌棉签，小心地取出肠道，直到可见肾下主动脉。接下来，从肾下主动脉解剖结缔组织和脂肪约 0.5 厘米的部分，同时避免小血管向背侧撕裂。然后将一块盐水浸泡的橡胶手套条放在腹主动脉和腹部主静脉下方，同时用棉签擦去多余的液体。之后，在肾下腹部脉管系统的外罩上包装一块浸有0.5摩尔氯化钙的纱布10分钟。取下纱布，包装另一块用PBS溶液浸泡五分钟的纱布，以在主动脉外膜的C2中生成钙正磷酸盐晶体。然后小心地取下橡胶手套条和纱布。重置小鼠肠道后，用五零缝合缝合腹部切口和皮肤。接下来，将鼠标放在加热垫上，直到鼠标恢复意识。将小鼠再安置14天，并在手术后密切监测小鼠，并随后至少每天观察。手术14天后，在腹侧切开安乐死的小鼠胸腔和腹腔，切开右心房。通过心脏左心室用PBS缓冲液大量使用小鼠以去除主动脉中的血液，然后如手稿中所述用4%多聚甲醛灌注。在体视显微镜下小心去除外源组织和脂肪。然后收获主动脉并将其放入装有五毫升4%多聚甲醛的管中48小时。然后用昆虫针将主动脉固定在黑色蜡板上，并继续获取主动脉图像。应用正磷酸钙14天后，对主动脉的形态进行成像，以可视化腹主动脉瘤的形成。观察到正磷酸钙导致肾下腹主动脉扩张，并导致弹性纤维的急剧降解，如弹性蛋白断裂所示。使用蘸有生理盐水的无菌棉签去除肠道，直到可见肾下主动脉，然后从肾下主动脉中提取结缔组织和脂肪。总体而言，亚磷酸钙诱导小鼠腹主动脉瘤模型是探索腹主动脉瘤分子机制和治疗策略的一种快速且具有成本效益的方法，可以与其他模型并行应用，以充分模仿人类腹主动脉瘤的特征。

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

Aortic Ring Assay

主动脉环含量

Angiogenesis, the sprouting of blood vessels from preexisting vasculature is associated with both natural and pathological processes. Various angiogenesis ...

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 ...

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 ...

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 ...

A Mouse Model of Intestinal Partial Obstruction

小肠部分梗阻的小鼠模型

Intestinal obstructions, that impede or block peristaltic movement, can be caused by abdominal adhesions and most gastrointestinal (GI) diseases including ...

Simultaneous Measurements of Intracellular Calcium and Membrane Potential in Freshly Isolated and Intact Mouse Cerebral Endothelium

新鲜分离和内联小鼠脑内皮细胞内钙和膜电位的同时测量

Cerebral arteries and their respective microcirculation deliver oxygen and nutrients to the brain via blood flow regulation. Endothelial cells line the ...

Imaging Calcium Dynamics in Subpopulations of Mouse Pancreatic Islet Cells

小鼠胰腺胰岛细胞亚群中的钙动力学成像

Pancreatic islet hormones regulate blood glucose homeostasis. Changes in blood glucose induce oscillations of cytosolic calcium in pancreatic islet cells ...

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A

质谱分析，用于识别 EYFP 标记 CENP-A （EYFP-CENP-A） 的泛化

Studying the structure and the dynamics of kinetochores and centromeres is important in understanding chromosomal instability (CIN) and cancer ...

A Mouse Model of Lumbar Spine Instability

腰椎不稳定的鼠标模型

Intervertebral disc degeneration (IDD) is a common pathological change leading to low back pain. Appropriate animal models are desired for understanding ...