Name: murine model of central venous stenosis using aortocaval fistula with an outflow stenosis
Uploaded: 2019-07-11T13:00:00
Description: Watch this Scientific Journal Video about Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis at JoVE.com

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。

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
<ol><li>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)。</li>
	<li>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。</li>
	<li>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?...

<ol>
	<li>Dember, L. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+clopidogrel+on+early+failure+of+arteriovenous+fistulas+for+hemodialysis:+a+randomized+controlled+trial.">Effect of clopidogrel on early failure of arteriovenous fistulas for hemodialysis: a randomized controlled trial.</a> JAMA. 299 (18), 2164-2171 (2008).</li><li>Dixon, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+don't+fistulas+mature?.">Why don't fistulas mature?.</a> Kidney International. 70 (8), 1413-1422 (2006).</li><li>Wilmink, T., Hollingworth, L., Powers, S., Allen, C., Dasgupta, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Natural+History+of+Common+Autologous+Arteriovenous+Fistulae:+Consequences+for+Planning+of+Dialysis.">Natural History of Common Autologous Arteriovenous Fistulae: Consequences for Planning of Dialysis.</a> European Journal of Vascular and Endovascular Surgery. 51 (1), 134-140 (2016).</li><li>Al-Jaishi, A. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patency+rates+of+the+arteriovenous+fistula+for+hemodialysis:+a+systematic+review+and+meta-analysis.">Patency rates of the arteriovenous fistula for hemodialysis: a systematic review and meta-analysis.</a> American Journal of Kidney Diseases. 63 (3), 464-478 (2014).</li><li>Rocco, M. V., Bleyer, A. J., Burkart, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Utilization+of+inpatient+and+outpatient+resources+for+the+management+of+hemodialysis+access+complications.">Utilization of inpatient and outpatient resources for the management of hemodialysis access complications.</a> American Journal of Kidney Diseases. 28 (2), 250-256 (1996).</li><li>Roy-Chaudhury, P., Sukhatme, V. P., Cheung, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemodialysis+vascular+access+dysfunction:+a+cellular+and+molecular+viewpoint.">Hemodialysis vascular access dysfunction: a cellular and molecular viewpoint.</a> Journal of the American Society of Nephrology. 17 (4), 1112-1127 (2006).</li><li>Quencer, K. B., Arici, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arteriovenous+Fistulas+and+Their+Characteristic+Sites+of+Stenosis.">Arteriovenous Fistulas and Their Characteristic Sites of Stenosis.</a> AJR: American Journal of Roentgenology. 205 (4), 726-734 (2015).</li><li>Kian, K., Asif, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cephalic+arch+stenosis.">Cephalic arch stenosis.</a> Semin Dial. 21 (1), 78-82 (2008).</li><li>Agarwal, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+vein+stenosis.">Central vein stenosis.</a> American Journal of Kidney Diseases. 61 (6), 1001-1015 (2013).</li><li>Glanz, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axillary+and+subclavian+vein+stenosis:+percutaneous+angioplasty.">Axillary and subclavian vein stenosis: percutaneous angioplasty.</a> Radiology. 168 (2), 371-373 (1988).</li><li>Yamamoto, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mouse+aortocaval+fistula+recapitulates+human+arteriovenous+fistula+maturation.">The mouse aortocaval fistula recapitulates human arteriovenous fistula maturation.</a> American Journal of Physiology: Heart and Circulatory Physiology. 305 (12), H1718-H1725 (2013).</li><li>North American Symptomatic Carotid Endarterectomy Trial. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods,+patient+characteristics,+and+progress.">Methods, patient characteristics, and progress.</a> Stroke. 22 (6), 711-720 (1991).</li><li>Kuwahara, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD44+Promotes+Inflammation+and+Extracellular+Matrix+Production+During+Arteriovenous+Fistula+Maturation.">CD44 Promotes Inflammation and Extracellular Matrix Production During Arteriovenous Fistula Maturation.</a> Arteriosclerosis, Thrombosis, and Vascular Biology. 37 (6), 1147-1156 (2017).</li><li>Protack, C. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eph-B4+regulates+adaptive+venous+remodeling+to+improve+arteriovenous+fistula+patency.">Eph-B4 regulates adaptive venous remodeling to improve arteriovenous fistula patency.</a> Scientific Reports. 7 (1), 15386 (2017).</li><li>Payne, H., Brill, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stenosis+of+the+Inferior+Vena+Cava:+A+Murine+Model+of+Deep+Vein+Thrombosis.">Stenosis of the Inferior Vena Cava: A Murine Model of Deep Vein Thrombosis.</a> J Vis Exp. (130), (2017).</li><li>Nam, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+carotid+ligation+is+a+model+of+acutely+induced+disturbed+flow,+leading+to+rapid+endothelial+dysfunction+and+atherosclerosis.">Partial carotid ligation is a model of acutely induced disturbed flow, leading to rapid endothelial dysfunction and atherosclerosis.</a> American Journal of Physiology: Heart and Circulatory Physiology. 297 (4), H1535-H1543 (2009).</li><li>Ene-Iordache, B., Remuzzi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disturbed+flow+in+radial-cephalic+arteriovenous+fistulae+for+haemodialysis:+low+and+oscillating+shear+stress+locates+the+sites+of+stenosis.">Disturbed flow in radial-cephalic arteriovenous fistulae for haemodialysis: low and oscillating shear stress locates the sites of stenosis.</a> Nephrology, Dialysis, Transplantation. 27 (1), 358-368 (2012).</li><li>Yamamoto, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disturbed+shear+stress+reduces+Klf2+expression+in+arterial-venous+fistulae+in+vivo.">Disturbed shear stress reduces Klf2 expression in arterial-venous fistulae in vivo.</a> Physiological reports. 3, (2015).</li><li>Remuzzi, A., Ene-Iordache, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+paradigms+for+dialysis+vascular+access:+upstream+hemodynamics+and+vascular+remodeling+in+dialysis+access+stenosis.">Novel paradigms for dialysis vascular access: upstream hemodynamics and vascular remodeling in dialysis access stenosis.</a> Clinical Journal of the American Society of Nephrology. 8 (12), 2186-2193 (2013).</li></ol>

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?...

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。

<table>
	<tbody>
		<tr>
			<td>20-60 Mhz scan head</td>
			<td>VisualSonics Inc.</td>
			<td>RMV-704</td>
			<td></td>
		</tr>
		<tr>
			<td>8-0 Sterile Micro Suture, 6mm (140 &#181;), 3/8 Circle, TAP Point Needle</td>
			<td>AROSuture</td>
			<td>T06A08N14-13</td>
			<td>polyamide monofilament sutures</td>
		</tr>
		<tr>
			<td>Induction Chamber, 2 Liter 
			3.75&#34;W x 9.00&#34;D x 3.75&#34;H</td>
			<td>VetEquip</td>
			<td>941444</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflo, Isoflurane liquid</td>
			<td>Zoetis</td>
			<td>26675-46-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Mice, C57BL/6J</td>
			<td>The Jackson Laboratory</td>
			<td>664</td>
			<td></td>
		</tr>
		<tr>
			<td>Pet Bed Microwave Heating Pad</td>
			<td>Snuggle Safe</td>
			<td>6250</td>
			<td></td>
		</tr>
		<tr>
			<td>PrecisionGlide Needle 25G</td>
			<td>BD</td>
			<td>305122</td>
			<td></td>
		</tr>
		<tr>
			<td>Surflo I.V. Catheter 22G</td>
			<td>Terumo</td>
			<td>SR-OX2225CA</td>
			<td>0.85mm outer diameter</td>
		</tr>
		<tr>
			<td>Vascular clamp</td>
			<td>Roboz Surgical Instrument</td>
			<td>RS-5424</td>
			<td></td>
		</tr>
		<tr>
			<td>Vevo770 High Resolution Imaging System</td>
			<td>VisualSonics Inc.</td>
			<td>770</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine model of central venous stenosis using aortocaval fistula with an outflow stenosis

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。

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?...

Watch this Scientific Journal Video about Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis at JoVE.com

Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis

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。

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

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ...

中央静脉狭窄可引起主动脉瘤，或AVF，成熟的主要失败，以及后来在工作管病的失败。此技术使用已建立的 murine AVF 模型的修改版本，该模型可重述人类 AVF 的临床过程，并通过几个关键步骤轻松掌握。在开始手术之前，确认在9至11周大的C57-Black/6小鼠中对手趾捏没有反应，并将鼠标放在手术区域的上位。将软膏涂抹在动物的眼睛上，将皮毛从颈部的腹侧取出至下腹部。然后用三个连续的10%的波维酮碘和70%的异丙醇磨砂对手术场所进行清洁和消毒，并在动物上放置手术窗帘。对于夹紧和穿刺部位暴露，将鼠标置于解剖显微镜下，并使用无菌技术和手术刀，从下肝边缘到上方进行皮肤深、中线、腹部切口。用剪刀切开肌肉打开腹腔，并将缩回器插入腹部。将肠子移动到右侧，用盐水浸泡的纱布包裹。将膀胱和显性囊泡移动到动物的骨侧，并使用微需要支架解剖直肠和直肠之间的肠胃，以确保大动脉和劣质的维纳卡瓦，或 IVC 的完整视图。然后使用微针架从横向和背部的逆白体组织解剖下肾大动脉和 IVC en 组，将组织交叉夹紧在一起，并解剖周围组织，在大约四分之三的距离处从左肾静脉到大动脉分叉暴露大动脉穿刺位点。对于 IVC 的隔离，在红外 IVC 和大动脉之间解剖，立即向左肾静脉进行解剖，将解剖分散到左肾静脉和大动脉分叉之间的一半，用于术后 IVC 观察。制作一个窗口，将 IVC 与大动脉分离，并解剖 IVC 与周围组织。然后小心地放置一个 8 - 0在通过窗口拉缝合线之前，在 IVC 和大母线下方的聚酰胺单丝缝合线仅将缝合线放在 IVC 下方的位置。要创建 AVF，请先将 25 量针弯曲到 45 到 60 度角，从针尖到针尖 4 毫米，然后用显微手术夹住下神经大母和 IVC。抓住分叉周围的结缔组织，以医疗方式和骨面旋转大母，将大母骨的穿刺位点稍微伸向腹侧，并使用 25 量针将大阳高刺入 IVC。当组织被刺穿时，释放大母，从大母骨左侧拉起，用周围组织覆盖穿刺位点。然后取出针头，然后用棉线棉签轻轻按压穿刺点，以进行血小球。对于 IVC 狭窄创建，将 22 量静脉导管的尖端纵向放置到 IVC 上，然后用 8-0 将导管粘附到 IVC缝合。放置缝合线后，取出导管并确认原位性异位，然后松开主干和 IVC。将穿刺位点再盖住一分钟，以确保血肿，然后将器官返回到腹腔，然后用6-0缝合关闭腹部。然后将鼠标放入热支持装置上的单个无床上用品笼中，并监控直至完全恢复，并使用多普勒超声波确认膀子的触度。在手术后的第七天，在纵向视图中很容易检测到 IVC 的管病和狭窄区域。在没有 AVF 的狭窄小鼠中，狭窄部分表现出静脉波形，其光谱比假操作的小鼠更宽，但没有太多的脉动性。在患有AVF和狭窄症的小鼠中，狭窄部分显示除光谱扩大外，脉动波形，而患有狭窄性AVF的小鼠在狭窄性手术中流动的时间平均最大速度明显高于单是患有狭窄症的小鼠。单单是狭窄小鼠的狭窄平均IVC直径与患有IVF和狭窄小鼠的均值IVC直径相似。此外，以上游或下游段为参考，除狭窄外，在患有 AVF 的小鼠中，狭窄百分比显著更大。当您将 IV 导管和 IVC 合二为一时，将它们紧密地绑起来以形成可重复的狭窄非常重要。本程序在静脉移植植入模型中的应用，可以回答如何克服静脉移植狭窄闭塞的问题。

Research

JoVE Journal

Medicine

Surgical Management of Meatal Stenosis with Meatoplasty

管理与Meatoplasty尿道外口狭窄的外科治疗

Meatal stenosis is a common urologic complication after circumcision.  Children present to their primary care physicians with complaints of deviated ...

科研

医学

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

采用实时定量PCR检测供体细胞植入一个具有竞争力的小鼠骨髓移植模型

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Technical Aspects of the Mouse Aortocaval Fistula

在的鼠标Aortocaval瘘的技术方面

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

动静脉内瘘失败的新型小鼠模型：在详细介绍手术过程

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Left Atrial Stenosis Induced Pulmonary Venous Arterialization and Group 2 Pulmonary Hypertension in Rat

左心房狭窄致大鼠肺静脉动脉化及第2组肺动脉高压

The mechanism of mitral stenosis-induced pulmonary venous arterialization and group 2 pulmonary hypertension (PH) is unclear. There is no rodent model of ...

A Protocol to Set Up Needle-Free Connector with Positive Displacement on Central Venous Catheter in Intensive Care Unit

在重症监护病房的中央静脉导管上设置无针接头,带正位移

Needle-free connectors were initially designed and promoted to avoid blood exposure for healthcare workers. Some recent data suggest that the latest ...

A Modified Two Kidney One Clip Mouse Model of Renin Regulation in Renal Artery Stenosis

肾动脉狭窄肾素调节的改良双肾单夹小鼠模型

Renal artery stenosis is a common condition in patients with coronary or peripheral vascular disease where the renin angiotensin aldosterone system (RAAS) ...

A Rabbit Aortic Valve Stenosis Model Induced by Direct Balloon Injury

球囊直接损伤诱导的兔主动脉瓣狭窄模型

Animal models are emerging as an important tool to understand the pathologic mechanisms underlying aortic valve stenosis (AVS) because of the lack of ...

Development of an Effective Mice Model for Studying Oronasal Fistulas

Establishment of an Oronasal Fistula Mice Model

Investigating Wound Healing in Mice Models of Oronasal Fistulas (Video) | JoVE

口鼻瘘小鼠模型的建立

This study presents a method utilizing heated ophthalmologic cautery to develop a viable model for investigating oronasal fistulas. C57BL/6 mice were used ...