中央静脉狭窄病的Murine模型,使用具有流出狭窄病的动脉静脉瘘管病
1Department of Surgery,Yale University, 2Vascular Biology and Therapeutics Program,Yale University, 3Department of Vascular Surgery,University of Tokyo, 4Department of Diagnostic Radiology,Tokai University School of Medicine, 5Department of Cardiovascular Surgery,Tokyo Medical University, 6Department of Vascular Surgery,VA Connecticut Healthcare Systems
Summary
主动脉瘘瘘是通过两面壁将马氏肾主动脉刺穿到劣质的vena卡瓦中,然后通过劣质vena卡瓦的部分结扎在流出中产生狭窄。这种可重现的模型可用于研究中央静脉狭窄。
Abstract
中静脉狭窄是导致动脉瘘 (AVF) 衰竭的重要实体。修改了鼠AVF模型,在瘘管流中创建劣质vena Cava(IVC)的部分结扎,模仿中央静脉狭窄。介绍了该模型的技术方面。主塔和IVC在腹部切口后暴露。对肾上腺主数和IVC进行解剖进行近端夹紧,并暴露远端主塔进行穿刺。左肾静脉和主动脉分叉之间的中点的IVC被仔细解剖,放置一个8-0缝合下IVC。夹紧主盘和IVC后,通过25G针将肾上腺主盘穿过两壁,将22G静脉内(IV)导管和IVC连在一起,从而形成AVF。然后,导管被移除,创建一个可重复的静脉狭窄,没有遮挡。主数和IVC在确认原发性赫利塞后未夹紧。这种新型的中央静脉狭窄模型易于执行,可重现,有利于对AVF故障的研究。
Introduction
与移植物或中央静脉导管等其他途径相比,动脉瘘 (AVF) 是血液透析最常见的访问途径,具有卓越的治疗率和减少感染。然而,高达60%的AVF不能成熟1,2,3;最近一项系统检讨报告,1年一级的评分率只有60%4。静脉流出的狭窄主要导致AVF成熟5、6的失效。有一些特征位置容易狭窄接近瘘管:并列的摆动段为放射性头瘘,头拱区域为胸脑直管瘘和中央静脉的瘘管与以前放置 ipsi侧子克隆或内静脉导管7,8。
中静脉狭窄往往是无症状的患者没有AVF,但可能导致静脉高血压的侧肢水肿,以及瘘管成熟失败时,挑战瘘管9。中央静脉狭窄的病理生理学最有可能与炎症和设备放置后激活凝血级联有关。此外,导管尖端的恒定运动以及瘘管流增加会改变剪切应力,导致血小板沉积和静脉壁变厚10。为了理解由中央静脉狭窄引起的AVF故障的基本机制,需要一种动物模型来模仿AVF的中央静脉狭窄。
我们建立了一个马腔瘘模型,易于执行,掌握和重述人类AVF的临床过程。11我们应用了几种先前建立的鼠模型的概念和技术,创建了具有静脉狭窄的新型鼠AVF模型。我们引入了一个在流出瘘中带有 IVC 狭窄的鼠主动脉瘘模型,可用于研究中央静脉狭窄。
Protocol
所有实验都是经耶鲁大学动物护理和使用委员会(IACUC)批准进行的。 1. 麻醉和术前程序 通过高压灭菌来消毒所有手术器械和材料。打开热支持装置,确保其温度(40~42 °C)。 将一只9-11周的旧C57BL/6小鼠放入丙烯酸感应室,用蒸发的2.5%等值胶和0.8升/分钟氧气将其麻醉。麻醉诱导大约需要3分钟。 将鼠标从造型室中取出。通过脚趾捏、耳捏和尾部捏合确认深层麻?…
Representative Results
雄性小鼠接受了上述手术,以产生AVF和IVC狭窄。对照小鼠只接受腹腔切除术和解剖IVC周围的组织,例如,假程序,或只创建IVC狭窄,而不同时创建AVF。 在手术后第7天,用多普勒超声波观察IVC(图2)。在纵向视图中很容易检测到 IVC 的瘘管和狭窄区域(图 2C,E)。瘘管和狭窄之间的IVC?…
Discussion
鼠AVF模型已用于研究导致AVF成熟13、14的基本机制和分子事件。在这项研究中,我们修改了一个已建立的鼠AVF模型,以创建一个新的鼠主动脉瘘模型,在瘘管流出区有IVC狭窄。我们的结扎模型类似于之前使用血管连接的几个小鼠模型。使用30G针垫15的局部IVC结扎,建立了深静脉血栓形成鼠模型;我们使用更大的22G IV导管间隔器,以创建一个较小?…
Declarações
The authors have nothing to disclose.
Acknowledgements
这项工作得到了美国国家卫生研究院(NIH)授予R01-HL128406的支持;美国退伍军人事务部生物医学实验室研究与发展计划优秀评审奖I01-BX002336;以及资源和设施的使用在VA康涅狄格州医疗保健系统,西黑文,CT。
Materials
| 20-60 Mhz scan head | VisualSonics Inc. | RMV-704 | |
| 8-0 Sterile Micro Suture, 6mm (140 µ), 3/8 Circle, TAP Point Needle | AROSuture | T06A08N14-13 | polyamide monofilament sutures |
| Induction Chamber, 2 Liter 3.75"W x 9.00"D x 3.75"H |
VetEquip | 941444 | |
| Isoflo, Isoflurane liquid | Zoetis | 26675-46-7 | |
| Mice, C57BL/6J | The Jackson Laboratory | 664 | |
| Pet Bed Microwave Heating Pad | Snuggle Safe | 6250 | |
| PrecisionGlide Needle 25G | BD | 305122 | |
| Surflo I.V. Catheter 22G | Terumo | SR-OX2225CA | 0.85mm outer diameter |
| Vascular clamp | Roboz Surgical Instrument | RS-5424 | |
| Vevo770 High Resolution Imaging System | VisualSonics Inc. | 770 |
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Citar este artigo
Isaji, T., Ono, S., Hashimoto, T., Yamamoto, K., Taniguchi, R., Hu, H., Wang, T., Koizumi, J., Nishibe, T., Hoshina, K., Dardik, A. Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis. J. Vis. Exp. (149), e59540, doi:10.3791/59540 (2019).