血管球囊损伤和腔内管理在大鼠颈动脉
Summary
该协议使用球囊导管引起的大鼠颈动脉腔内损伤，从此引起内膜增生。这是一个公认的模型用于研究在响应损伤的血管重塑的机制。它也被广泛使用，以确定潜在的治疗方法的有效性。
Abstract
大鼠颈动脉球囊损伤模型已经确立了二十多年。它仍然是学习与之相关的血管平滑肌分化，形成内膜和血管重塑的分子和细胞机制的重要方法。雄性Sprague-Dawley大鼠是最经常使用的动物为这种模式。雌性大鼠不优选的，因为雌激素是保护性防止血管疾病，因而引入一个变化成此过程。左颈通常受伤与右颈动脉作为阴性对照。左颈损伤是由膨胀的气球是滥伐内皮和扩张S中的血管壁造成的。损伤后，潜在的治疗策略，例如使用药物化合物和任一基因或shRNA转移的进行评估。典型地，对于基因或shRNA转移，血管腔的受伤部分被局部转30分钟，用viral粒子交付和表达了受伤的血管壁或者编码蛋白质或shRNA。新内膜增厚较增殖的血管平滑肌细胞通常损伤后峰值在2周。船只大多收获用于细胞和分子的细胞信号转导途径的分析以及基因和蛋白表达该时间点。容器也可收获在较早时间点，以确定一个特定的蛋白质或途径的表达和/或活化的发作，这取决于预期的实验目的。容器可以被表征和评价使用组织染色，免疫组织化学，蛋白质/表达测定法，和活性测定。来自同一动物的完整右颈动脉是一种理想的内部控制。在分子和细胞损伤的参数引起的变化可以通过动脉受伤比较内部右控制动脉进行评估。同样地，治疗方法可以通过比较损伤进行评估D和治疗动脉控制只受伤的动脉。
Introduction
气囊导管在血管成形术的过程中使用，以扩大或动脉粥样硬化血栓阻塞网站，在血管的目的的医疗器械。变窄的血管腔被强制由膨胀的气球，以开放和血液供应将被顺序地恢复到解除下游局部缺血症状，例如心绞痛，心肌梗塞，和腿痛。然而，血管成形术的巨大成功已经减少了术后并发症如力结果引起血管气压伤（球囊损伤），即血管壁重塑，在很多情况下，再狭窄的血管腔（再狭窄）1。
已经开发了许多动物模型模仿血管成形术，以帮助研究人员了解球囊损伤有关的血管壁重塑2背后的机制。在所有的用于建模的动物物种，鼠是最经常使用的一个。 Çompared到兔，狗和猪，鼠的优势是其成本低，其相对的易用性和大鼠生理学的现有知识。虽然小鼠具有一个额外的优势在广泛基因操作的菌株，小鼠容器太小，插入气囊导管。在过去的三十年里，实验组大鼠已使研究人员能够更好地了解支撑内膜增生及血管重塑3-6的分子和细胞机制。超越球囊损伤，血管重塑也参与大多数主要血管疾病，如动脉粥样硬化7,8，高血压9和动脉瘤10。因此，知识通过球囊损伤模型，获得了在一般的整体血管壁病变的研究是有益的。
大鼠球囊损伤模型的总体目标是不仅要进一步理解的血管疾病，而且，以测试新的药剂的效力为疾病控制11,12。目前临床药物治疗再狭窄是由血管成形术后右经血管腔放置药物洗脱支架应用。在动物模型中，一个高效而更经济的方式为新代理的测试是一个发达的地方腔内灌注的方法。已通过这种方法进行了测试候选药物包括小分子药物13,14，细胞因子或生长因子15,16，基因操纵剂（cDNA克隆，的siRNA 等 ）17-20，以及新颖的药物制剂21,22。
到目前为止，大鼠球囊损伤模型仍然是研究血管疾病/病症的最有用的模型之一。它是从台架的基本步骤到床头，通常作为第一步骤，从体外移动到体内 ，但它不应该是最后一个。大鼠实验的结果需要被审议和进一步的特征在于转换到人体之前临床使用时，由于在血管床和容器解剖学以及人和大鼠23-26之间的内在物种差异的差异。尽管如此，它仍然是在转化医学研究的重要工具。虽然这种研究用于因缺乏转基因鼠的限制，它已不再是一个问题，因为新的基因组的方法，如锌指核酸酶27，TALENS 28和CRISPR-CAS 29取得淘汰赛大鼠方便。
Protocol
Representative Results
Discussion
大鼠颈动脉球囊损伤一直深受Tulis在2007年34说明，它已全面通过Tulis博士讨论了此过程的所有细节。谁是有意执行此过程的读者强烈推荐阅读Tulis“协议。然而，有一件事我们不Tulis博士同意：除了充气用生理盐水或任何液体的气球，我们建议与空气膨胀了。根据我们的个人经验，液体膨胀难以避免气泡。此外，它更难以灵活调整损伤过程中的压力，并可能导致额外的压力和伤害到动脉。另一个技术技巧是…
Declarações
The authors have nothing to disclose.
Acknowledgements
We are grateful to Dr. Clowes for first developing and describing this method. We are also thankful to Dr. Tulis for his detailed protocol which has been fundamentally helpful to our previous, current and future work. This work was supported by grants R01HL097111 and R01HL123364 from the NIH to M.T., and by American Heart Association grant 14GRNT18880008 to M.T.
We would like to thank Rachel Newton for her expert technical support and for her valuable help during the filming process.
Materials
| Name of Material/ Equipment | Company | Catalog Number | Comments/Description |
| Fogarty balloon embolectomy catheters, 2 French | Edwards Lifesciences, Germany | 120602F | |
| Deltaphase Operating Board – Includes 2 Pads & 2 Insulators | Braintree Scientific, Inc. | 39OP | |
| LED light source | Fisher Scientific | 12-563-501 | |
| Hartmann Mosquito Forceps 4” curved | Apiary Medical, Inc. San Diego, CA | gS 22.1670 | |
| Crile Retractor 4” double ended | Apiary Medical, Inc. | gS 34.1934 | |
| Other surgical instruments | Roboz Surgical Instrument Company, Inc., Gaithersburg, MD | ||
| Peripheral Intravenous (I.V.) Cannula, 24G | BD | 381312 | |
| Ketamine HCl, 100mg/mL, 10mL | Ketaset- Patterson Vet | 07-803-6637 | |
| Xylazine (AnaSed),20mg/mL,20mL | Ketaset- Patterson Vet | 07-808-1947 | |
| Buprenex, 0.3mg/1ml (5 Ampules/Box) | Ketaset- Patterson Vet | 07-850-2280 | |
| Nair Baby Oil Hair Removal Lotion-9 oz | Amazon/Walmart/CVS | N/A | |
| Inflation Device | Demax Medical | DID30 | |
| D300 3-way Stopcock | B.Braun Medical Inc. | 4599543 | |
| Artificial Tears Ointment | Rugby Laboratories, Duluth, GA | N/A |
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