隔离和小鼠主动脉切除;心血管疾病研究的一个多功能的技术
Summary
Pathology of the aorta can lead to severe morbidity and mortality, therefore research of disease progression and potential therapies is warranted. Here, we present a protocol to isolate and excise the murine aorta to aid researchers in their investigation of cardiovascular disease.
Abstract
Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated blood is the aorta. The aorta may become affected in diseases such as atherosclerosis and aneurysm. Researchers investigating these diseases would benefit from direct observation of the aorta to characterize disease progression as well as to evaluate efficacy of potential therapeutics. The goal of this protocol is to describe proper isolation and excision of the aorta to aid investigators researching cardiovascular disease. Isolation and excision of the aorta allows investigators to look at gross morphometric changes as wells as allowing them to preserve and stain the tissue to look at histologic changes if desired. The aorta may be used for molecular studies to evaluate protein and gene expression to discover targets of interest and mechanisms of action. This technique is superior to imaging modalities as they have inherent limitations in technology and cost. Additionally, primary isolated cells from a freshly isolated and excised aorta can allowing researchers to perform further in situ and in vitro assays. The isolation and excision of the aorta has the limitation of having to sacrifice the animal however, in this case the benefits outweigh the harm as it is the most versatile technique in the study of aortic disease.
Introduction
主动脉在心血管功能的关键作用供给体与含氧血液和身体的类似其他部分，是容易感染疾病。常见的主动脉疾病包括动脉粥样硬化和动脉瘤，这是遗传和环境因素，包括饮食，吸烟和久坐的生活方式1的结果。动脉粥样硬化是一种碳酸钙和基于脂质的斑块到主动脉壁的通常存在于患有慢性血症2的沉积。动脉瘤的特征是主动脉和血管壁的变薄的结构部件，接着是直径增加最终会出现破裂3的降解。
动物模型是用于研究疾病和潜在的治疗功效的机制的重要工具。在心血管研究中常用的动物模型，专门研究调查血管及代谢紊乱包括转基因小鼠，以改变脂质水平，如载脂蛋白E基因敲除和低密度脂蛋白受体基因敲除小鼠4。多数方法来评估疾病和治疗功效的机制将包括主动脉的分离和切除。
一旦主动脉被定位和分离，形态测定分析可确定，如存在动脉瘤，通常定义为在直径5增加大于50％。经过必要的原位分析完成后，主动脉可切下用于进一步分析。一个切除的主动脉，可快速冷冻以进行分子学研究，如蛋白质和/或基因表达的测定法或用4％多聚甲醛固定和更高包埋，切片并染色用于组织学分析。主动脉的组织学分析，可以显示主动脉疾病的共同特征，例如结构退化，斑形成，和白细胞浸润<SUP> 6,7。此外，一个离体主动脉可用于分离原代细胞系，包括内皮细胞和平滑肌细胞，随后可以用于各种体外研究7。
目前，还没有其他的方法来提供这种深入表征主动脉疾病，以及为研究人员提供工具，进一步研究心血管疾病。成像模态诸如磁共振成像，计算机断层扫描和超声是形态学评估的主动脉的接近的方法，然而，这是难以在小动物和获取设备与适当的技术是昂贵的8。永生化细胞系可购到调查疾病和疗法的效力的潜在机制，但是这些细胞的人造性质被限制在研究对细胞周期和凋亡9的影响。
的总目标该原稿是展示在心血管疾病的调查鼠主动脉的无菌隔离和切除。
Protocol
Representative Results
Discussion
Aortic disease can lead to significant systemic pathology and possibly death in severe cases. Treatment options are currently limited therefore continued research in the field is necessary10. Murine models are ideal to further study disease progression and potentially therapeutic options4. Therefore, the successful completion of the method described in this manuscript will provide investigators with the tools to measure disease progression and drug efficacy.
This techniq…
Disclosures
The authors have nothing to disclose.
Acknowledgements
作者没有确认。
Materials
| Name of Reagent/ Equipment | Company | Catalog Number | Comments/Description |
| Isoflurane | Med-Vet International | #RXISO-250 | |
| 70% Ethanol | Fisher | 07-678-001 | |
| Phosphate buffered saline | Sigma Aldrich | P5368-10PAK | |
| Surgical Tape | 3M | 1527-1 | |
| Sterile Gauze | Dukal Corporations | 1312 | |
| 25 Gauge Needle | BD | 305122 | |
| 10ml Syringe | BD | 309604 |
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