大鼠颈动脉球囊损伤模型试验抗血管重建治疗
Summary
The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.
Abstract
The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.
Introduction
血管成形术是用于加宽狭窄或阻塞的动脉从病理状况例如动脉粥样硬化所得的血管内手术。血管成形术的一种常见并发症是后期运营内膜增生，或再狭窄，发生由于手术伤害和随后的炎症引起的血管重塑。这些条件导致血管平滑肌细胞和多种病理生理后果1-3的增殖。新内膜增生再变稠容器中，并发生在高达60％的血管成形术后的患者的第一年之内。因此，再狭窄是广泛使用的血管成形术4的重大挫折。虽然药物洗脱支架植入可能有助于防止再狭窄，唯一入选的考生可以接受这种昂贵的步骤5。
动物和临床研究中已经确定，通过vascu产生慢性炎症LAR伤害和/或手术伤口用作血管成形术后内膜生长2,4主要刺激。大鼠颈动脉损伤模型模拟临床情况，因此作为一个有价值的模型系统识别，在血管重构和血管细胞增殖6-9涉及的细胞因子。该模型系统也是评价一个非常有用的工具和/或屏幕为抑制在临床前翻译研究10-14新内膜生长的药物和治疗的试剂。
相较于小鼠颈动脉导线损伤模型15和小鼠股动脉导线损伤模型16，大鼠颈球囊损伤模型中是规模足够大，便于手术操作，便于对造成的伤害的可重复性的优势。它可以提供原代细胞为ADDIT更大数目（ 例如 ，血管平滑肌细胞，内皮细胞）有理体外研究来描绘理事血管重塑的分子机制。重要的是，相对于小鼠，大鼠也已知可用于生理和毒理学研究17更好的模型。虽然大鼠模型的缺点或限制是缺乏遗传改良和基因敲除模型，这个缺点可以通过大鼠基因组序列的可用性以及最近强大的基因组编辑工具，如CRISPR-CAS技术，使发展来克服在不同的模型系统18,19基因组序列的广大范围可能操纵。
虽然大鼠球囊损伤模型已被用于多个实验室和各种综合性协议已经公布的20,21，这个协议的目的是在手术前的准备工作，提供更多的细节，可以引导研究人员新的这个步骤设置这种手术的做法。我们还强调，手术后护理的t他的动物，允许未对动脉重塑的治疗效果只有验尸病理组织形态学和分析，也超声声像图研究活体动物13,22。
Protocol
Representative Results
Discussion
已经有用于充气气球以产生，消除在颈动脉管腔壁画内皮损伤的两种方法。之一是用液体20填充附注射器，而另一种是使用空气压力21。我们更喜欢使用液体填充的注射器，因为确切的液体体积（0.02毫升）将用于每个过程。这使得该气球的精确和可重复的通货膨胀，导致损伤的每只动物接受的过程的类似水平。在注射器中使用的液体确实需要在液体去除气泡。这可以通过从导管的?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
The work was supported by the intramural research program of the NIH, National Institute on Aging, and by a Priority Research Centers Program grant from the National Research Foundation (NRF-2009-0093812) funded by the Ministry of Science, Information and Communication Technology, & Future Planning, the Republic of Korea (H.T.). We thank Dr. Han-sol Park for putting “material” part together for the manuscript.
Materials
| 2 F Fogarty balloon embolectomy catheter | Edwards Lifesciences |
| Standard scalpel | Fine Science Tools |
| Small curved forceps (Large radius Dumont#7shanks curved) | Fine Science Tools |
| Large, medium and small micro-scissors | Roboz |
| Needles (20 G) | TycoHealthcare |
| Micro-surgery forceps with micro-blunted atraumatic tips | Fine Science Tools |
| Atraumatic straight small arterial clamps | Fine Science Tools |
| Retractor with maximum spread 5.5 cm long blunt teeth | Fine Science Tools |
| Silk suture (4.0 and 6.0 ) | Fine Science Tools |
| Syringe (1.0 ml) | BD |
| Curity gauze sponges | AllegroMedical |
| Cotton tip applicators sterile and non-sterile | Puritan Medical Products |
| Compact hot bead sterilizer | Fine Science Tools |
| Self-regulating heating pad | Fine Science Tools |
| ADS200 anesthesia system/ventilator | Paragon Medical |
| Isoflurane (forane), liquid form | Baxter |
| Sodium chloride 0.9% (Saline) | Hospira |
| Buprenex (buprenorphine) | Reckitt Benckiser Healthcare (UK) Ltd. |
| 70% alcohol | Fisher |
| 1: 10 Betadine | Fisher |
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