磷酸钙诱导的小鼠腹主动脉瘤模型
Summary
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。
Abstract
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。
Introduction
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。
Protocol
Representative Results
Discussion
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?…
Acknowledgements
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。
Materials
| CaCl2 | MECKLIN | C805225 | |
| NaCl | Biomed | SH5001-01 | |
| PBS | HARVEYBIO | MB5051 | |
| Small animal ventilator | RWD | H1550501-012 |
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