再狭窄的鼠标股动脉线损伤模型在技术上具有挑战性。在这个协议中,我们展示的关键技术细节必不可少的成功执行丝损伤诱导一致内膜再狭窄的研究。
引起的动脉粥样硬化的心血管疾病是死亡的在发达国家的首要原因。变窄的血管腔,由于动脉粥样硬化斑块的发展或建立斑块的破裂,中断正常的血流导致各种并发症,如心肌梗塞和中风。在门诊血管内手术,如血管成形术通常进行重新管腔。然而,这些治疗方法不可避免地破坏血管壁以及血管内皮,触发过度愈合反应和新内膜斑块伸入引起血管再狭窄(再狭窄)的内腔的发展。再狭窄仍然是动脉粥样硬化的血管内治疗失败的主要原因。因此,再狭窄的临床前动物模型是用于研究的病理生理机制,以及平移接近血管介入至关重要。在几种鼠experimental模型,股动脉导线损伤被广泛接受作为用于血管成形术后再狭窄的研究最合适的,因为它非常类似于该损伤都内皮和血管壁的血管成形术。然而,许多研究人员利用该模型困难,由于其高的技术难度程度。这主要是因为金属线需要被插入到股动脉,这大约比丝更薄三次,以产生足够的损伤诱导突出的新内膜。在这里,我们描述了必要的手术细节,有效地克服了这一模式的主要技术难点。按照所提出的程序,在执行鼠标股动脉导线损伤变得更容易。一旦熟悉,整个过程可以在20分钟内完成。
In the era of expanded application of endovascular treatments for various cardiovascular diseases, restenosis after angioplasty is one of the major problems for patients undergoing such treatments. Damage to the vascular endothelium at the time of angioplasty, in concert with the atherosclerotic background, induces excessive smooth muscle cell proliferation in the medial layer, resulting in neointimal hyperplasia.1,2 A viable animal model that recapitulates post-angioplasty neointimal hyperplasia is, thus, important not only for the investigation of disease mechanisms but also for the development of effective therapeutics to treat this pathology.
Mice represent an excellent model animal to recapitulate neointimal hyperplasia for the following reasons: the genetic backgrounds of experimental mice are well established; a wide variety of genetically-modified strains are available;2 obtaining littermates of the same background is easy; and the cost of the animals is relatively low. Arterial ligation model and wire injury model are the two most common mouse models of mechanically-induced neointimal hyperplasia. The arterial ligation model is easy to create, but physiologically dissimilar to the actual angioplasty procedure. The wire injury model closely mimics actual angioplasty procedures but is technically difficult due to the small size of mouse arteries.2,3 Sata et al. first described a wire injury method for mouse femoral arteries based on the anatomical structure of the vasculature and the use of proper-sized flexible wire. Utilizing this technique, they succeeded in reproducibly inducing neointimal hyperplasia in various strains of mice.4
Although femoral wire injury is a well-established model, some of the technical aspects of the technique are highly challenging compared to other models such as ligation.5 The purpose of this paper is to describe our mouse wire injury model procedures in detail, which is a modified version of Sata’s original method. We have made two main modifications: 1) Looping only the arteries, and 2) No lidocaine use.
导线损伤程序适用于小鼠所有菌株,只要它们的解剖结构是相似的。4在本文中,我们使用雄性C57BL / 6小鼠的12-16周的年龄。正如我们从我们的初步研究中了解到,小鼠年龄小于10周的股动脉往往是如此之小,导线插入是相当具有挑战性的。另一方面,在超过16周小鼠丝插在技术上更容易,但倾向于导致更多变新内膜增生。雌性小鼠往往会产生更少的新生内膜形成较男性。虽然丝损伤过程可?…
The authors have nothing to disclose.
This work was supported by a Wisconsin Partnership Program New Investigator Award (ID 2832), a National Heart, Lung, Blood Institute R01 Grant (HL-068673) and a T32 training Grant (HL-110853). We thank Dr. Melina Kibbe’s group at Northwestern University for providing helpful information.
Name of Reagent/ Equipment | Company | Catalog Number | Comments/Description |
Fixed Core Wire Guide | Cook | G02426 | Diameter 0.015 inch, Straight |
Dilation Forceps | Roboz | RS-4927 | Curved, blunt tips |
Dumont Tweesers #5 | World Precision Instruments | 14095 | Straight, sharp tips |
Dumont Vessel Cannulation Forceps | World Precision Instruments | 503373 | |
McPherson-Vannas Scissors | World Precision Instruments | 501234 | |
Mosquito Forceps | World Precision Instruments | 501291 | |
Ethilon Nylon Suture 9-0 | Ethicon | 7717G | 9-0, Black Nylon Monofilament |
Micro AROSuture, Sterile 11-0, 70 Microns, MET Point | AROSurgical | VT4A00N07 | 11-0, Black Nylon Monofilament |
3M Precise Multi-Shot DS Disposable Skin Stapler | 3M | DS-25 |