Name: ferric chloride-induced canine carotid artery thrombosis: a large animal model of vascular injury
Uploaded: 2018-09-07T14:45:00
Description: Watch this Scientific Journal Video about Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury at JoVE.com

我们要感谢俄亥俄州立大学认知和行为脑成像中心的财政和科学支持, 以发展和执行犬磁共振成像。

所述调查符合《国家卫生研究院护理和使用实验动物指南》, 并获得俄亥俄州立大学机构动物保育和使用委员会 (#2015A00000029) 的批准。所有手术操作都是在深部麻醉下进行的, 在手术过程中, 动物在任何阶段都没有疼痛。所描述的所有实验都是不恢复的。
1. 准备
<ol><li>新鲜地准备50毫升氯化铁 (FeCl3) 在 50% (瓦特/v) 稀释在去离子水。</li>
	<li>将脐带切成5.5 厘米片, 浸泡?...

<ol>
	<li>Adams, H. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stroke:+a+vascular+pathology+with+inadequate+management.">Stroke: a vascular pathology with inadequate management.</a> Journal of Hypertension Supplement. 21 (5), S3-S7 (2003).</li><li>Lansberg, M. G., Bluhmki, E., Thijs, V. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+and+safety+of+tissue+plasminogen+activator+3+to+4.5+hours+after+acute+ischemic+stroke:+a+metaanalysis.">Efficacy and safety of tissue plasminogen activator 3 to 4.5 hours after acute ischemic stroke: a metaanalysis.</a> Stroke. 40 (7), 2438-2441 (2009).</li><li>Nagel, 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=Therapy+of+acute+basilar+artery+occlusion:+intraarterial+thrombolysis+alone+vs+bridging+therapy.">Therapy of acute basilar artery occlusion: intraarterial thrombolysis alone vs bridging therapy.</a> Stroke. 40 (1), 140-146 (2009).</li><li>Ciccone, A., Motto, C., Abraha, I., Cozzolino, F., Santilli, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycoprotein+IIb-IIIa+inhibitors+for+acute+ischaemic+stroke.">Glycoprotein IIb-IIIa inhibitors for acute ischaemic stroke.</a> The Cochrane database of systematic reviews. 3 (3), (2014).</li><li>The National Institute of Neurological Disorders and Stroke rt-PA Stroke Study Group. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+Plasminogen+Activator+for+Acute+Ischemic+Stroke.">Tissue Plasminogen Activator for Acute Ischemic Stroke.</a> New England Journal of Medicine. 333 (24), 1581-1588 (1995).</li><li>Leadley, R., Chia, L., Rebellob, S., Gagnon, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+in+vivo+models+of+thrombosis+to+the+discovery+and+development+of+novel+antithrombotic+agents.">Contribution of in vivo models of thrombosis to the discovery and development of novel antithrombotic agents.</a> Journal of Pharmacological and Toxicological Methods. 43 (2), 101-116 (2000).</li><li>Bodary, P. F., Eitzman, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+of+Thrombosis.">Animal Models of Thrombosis.</a> Current Opinion In Hematology. 16 (5), 342-346 (2009).</li><li>Sachs, U. J. H., Nieswandt, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+thrombus+formation+in+murine+models.">In vivo thrombus formation in murine models.</a> Circulation Research. 100 (7), 979-991 (2007).</li><li>Bonnard, T., Hagemeyer, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ferric+Chloride-induced+Thrombosis+Mouse+Model+on+Carotid+Artery+and+Mesentery+Vessel.">Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel.</a> Journal of Visualized Experiments. (100), e52838 (2015).</li><li>Kurz, K. D., Main, B. W., Sandusky, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+model+of+arterial+thrombosis+induced+by+ferric+chloride.">Rat model of arterial thrombosis induced by ferric chloride.</a> Thrombosis Research. 60 (4), 269-280 (1990).</li><li>Karatas, H., Erdener, S. E., 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=Thrombotic+distal+middle+cerebral+artery+occlusion+produced+by+topical+FeCl(3)+application:+a+novel+model+suitable+for+intravital+microscopy+and+thrombolysis+studies.">Thrombotic distal middle cerebral artery occlusion produced by topical FeCl(3) application: a novel model suitable for intravital microscopy and thrombolysis studies.</a> Journal of Cerebral Blood Flow and Metabolism. 31 (6), 1452-1460 (2011).</li><li>Li, W., McIntyre, T. M., Silverstein, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ferric+chloride-induced+murine+carotid+arterial+injury:+A+model+of+redox+pathology.">Ferric chloride-induced murine carotid arterial injury: A model of redox pathology.</a> Redox Biology. 1 (1), 50-55 (2013).</li><li>Vilahur, G., Padro, T., Badimon, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atherosclerosis+and+Thrombosis:+Insights+from+Large+Animal+Models.">Atherosclerosis and Thrombosis: Insights from Large Animal Models.</a> Journal of Biomedicine and Biotechnology. 2011, 1-12 (2011).</li><li>Coller, B. S., Folts, J. D., Smith, S. R., Scudder, L. E., Jordan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abolition+of+in+vivo+Platelet+Thrombus+Formation+in+Primates+with+Monoclonal+Antibodies+to+the+Platelet+GPIIb/IIIa+Receptor.+Correlation+with+Bleeding+Time,+Platelet+Aggregation,+and+Blockade+of+GPIIb/IIIa+Receptors.">Abolition of in vivo Platelet Thrombus Formation in Primates with Monoclonal Antibodies to the Platelet GPIIb/IIIa Receptor. Correlation with Bleeding Time, Platelet Aggregation, and Blockade of GPIIb/IIIa Receptors.</a> Circulation. 80 (6), 1766-1774 (1989).</li><li>Folts, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vivo+Model+of+Experimental+Arterial+Stenosis,+Intimal+Damage,+and+Periodic+Thrombosis.">An in vivo Model of Experimental Arterial Stenosis, Intimal Damage, and Periodic Thrombosis.</a> Circulation. 83 (6 Suppl), (1991).</li><li>Yasuda, T., 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=A+canine+model+of+coronary+artery+thrombosis+with+superimposed+high+grade+stenosis+for+the+investigation+of+rethrombosis+after+thrombolysis.">A canine model of coronary artery thrombosis with superimposed high grade stenosis for the investigation of rethrombosis after thrombolysis.</a> Journal of the American College of Cardiology. 13 (6), 1409-1414 (1989).</li><li>Schob, 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=Correlation+Between+Aquaporin+4+Expression+and+Different+DWI+Parameters+in+Grade+I+Meningioma.">Correlation Between Aquaporin 4 Expression and Different DWI Parameters in Grade I Meningioma.</a> Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging. 19 (1), 138-142 (2017).</li><li>Schob, 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=Diffusion-Weighted+Imaging+Using+a+Readout-Segmented,+Multishot+EPI+Sequence+at+3+T+Distinguishes+between+Morphologically+Differentiated+and+Undifferentiated+Subtypes+of+Thyroid+Carcinoma-A+Preliminary+Study.">Diffusion-Weighted Imaging Using a Readout-Segmented, Multishot EPI Sequence at 3 T Distinguishes between Morphologically Differentiated and Undifferentiated Subtypes of Thyroid Carcinoma-A Preliminary Study.</a> Translational Oncology. 9 (5), 403-410 (2016).</li><li>Schob, 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=Diffusion-Weighted+MRI+Reflects+Proliferative+Activity+in+Primary+CNS+Lymphoma.">Diffusion-Weighted MRI Reflects Proliferative Activity in Primary CNS Lymphoma.</a> Public Library of Science One. 11 (8), e0161386 (2016).</li></ol>

FeCl3诱导的血管损伤模型广泛用于研究小动物血栓形成, 易于转化为大型动物, 具有多种优势的临床前模型。将该协议调整为犬类的轻微修改, 允许利用磁共振成像对中风和出血量进行药理干预和血管造影评估, 评估血管疏导之前、期间和治疗后。其他血栓性大动物模型未研究损伤部位的稳定闭塞血栓, 因此不能利用颈动脉血管造影和组织学评估每种预防性或溶栓疏导的程度。治疗。除了大?...

中风治疗主要是在冠状动脉疾病治疗后进行模拟的, 因为心血管疾病的干预对药物治疗和血管内干预措施有很好的反应1。然而, 这些治疗并没有成功地转化为脑梗塞。目前中风治疗的难点是重组组织纤溶酶原激活剂 (rTPA) 不能逆转, 而且该管理具有6.4% 的出血转化率2,3的风险,4. 由此产生的发病率和死亡率限制了它对一个小的、常常无法实现的窗口5的使用。同时, 再狭窄和闭塞往往发生在最初的溶栓后, 逆转最初的神经系统改善。总之, 有一个狭窄的时间窗口管理 rTPA, 排除大多数 (90%) 的患者谁患有缺血性脑血管的侮辱。
静脉抗血小板治疗在改善血管再通、生存和结局2方面显示了治疗缺血性中风的前景。不幸的是, 这些药物有一个可预测的副作用的颅内和颅内出血, 主要是因为没有办法来充分逆转或控制他们的活动2。虽然有效的预防血小板聚集, 出血的风险和无法逆转他们的活动已排除在日常护理中风患者使用。因此, 有必要采取有效的抗血栓药物, 单独或组合, 以防止和溶解血栓还有一个安全的配置文件, 将允许使用在封闭的, 低体积的空间, 如大脑, 出血是不耐受的。
了解动脉血栓形成和再狭窄的机制, 评估预防再狭窄的栓和药物, 需要小动物模型作为临床前药物开发的一部分。氯化铁诱导的血管损伤是快速、准确诱导小鼠、大鼠 、豚鼠和家兔6、7、8等暴露血管血栓形成的一种广泛应用的技术,9,10,11,12. 这些较小的物种提供了一些好处, 包括易于遗传操作、廉价的动物购买和低的每日津贴住房费用。不幸的是, 小动物实验在手术中否定了多项血液绘制, 以获得血小板反应性、血气分析和炎症反应。更重要的是, 大型动物更紧密地模仿人类血小板生理学6,13。FeCl3型颈动脉损伤模型在研究血栓的病理生理学、新的抗血小板和抗凝血药物的有效性以及发现潜在栓6中发挥了主导作用。,7,8,9,10,11,12. 以前在小鼠、大鼠、豚鼠和兔子身上的模型为基因操作提供了方便和灵活性, 但可翻译的临床前模型对药物治疗药物的剂量和毒性研究有重要意义6 ,13。虽然在小鼠中已经形成了几种血栓紊乱模型, 但适用于周围血管疾病、中风和心肌梗塞的大动物血栓形成模型却很少。第一个血栓模型在猴子, 狗和猪集中于狭窄, 应用止血剂和以后圆筒对容器, 通常导致循环流减少14,15,16。而不是在内皮损伤部位的闭塞血栓, 如在氯化铁模型中, 这些模型中的血栓导致循环血栓形成, 远端栓塞, 恢复正常血流。相比之下, 在大动物这里修改的氯化铁模型, 导致损伤部位闭塞血栓, 并在溶栓治疗前通过血管造影进行稳定和验证。如果调查人员有足够的资金用于每日津贴和购买犬齿以及足够的外科专业技能, 我们这里详细介绍一大犬血管损伤模型, 让实验室通过手术、影像学和组织学研究血栓形成。技术。

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	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:397px;">1/8&#8221; umbilical tape&#160;</td>
			<td style="width:195px;">Jorgensen Laboratories Inc.,&#160;</td>
			<td style="width:344px;">#J0025UA&#160;</td>
			<td style="width:201px;">for ferric chloride application</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">4% paraformaldehyde in PBS</td>
			<td>Alfa Aesar</td>
			<td>AAJ61899AP</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">10% neutral buffered formalin&#160;</td>
			<td>Richard-Allan Scientific</td>
			<td>5701</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">&#160;2% 2,3,5-triphenyltetrazolium chloride (TTC in PBS, pH 7.4)&#160;</td>
			<td>Sigma Aldrich</td>
			<td>T8877</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">ADP/Collagen cartridges</td>
			<td>Siemens Diagnostics</td>
			<td>B417021A</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">4.5 ml 3.2% sodium citrate blood vacutainer&#160;</td>
			<td>Becton Dickinson</td>
			<td>BD 369714</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">4.5 ml lithium heparin vacutainer&#160;</td>
			<td>Becton Dickinson</td>
			<td>BD 368056</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">EDTA K3 vacutainers&#160;</td>
			<td>Becton Dickinson</td>
			<td>BD455036</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Doppler flow probe</td>
			<td>Transonic Systems Inc</td>
			<td>MA2.5PSL</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Hematoxylin 560&#160;</td>
			<td>Surgipath</td>
			<td>3801570</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Eosin</td>
			<td>Surgipath</td>
			<td>3801602</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">LabChart Software</td>
			<td>ADInstruments Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Prisma Fit 3 tesla (3T) magnet</td>
			<td>Siemen&#39;s Diagnostics</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Sodium heparin for injection (to coat blood gas syringe)</td>
			<td>NovaPlus</td>
			<td>402525D</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">HUG-U-VAC positioning system &#160;</td>
			<td>DRE Veterinary</td>
			<td>1320</td>
			<td></td>
		</tr>
	</tbody>
</table>

ferric chloride-induced canine carotid artery thrombosis: a large animal model of vascular injury

导致脑缺血性中风和心肌梗死的闭塞性动脉血栓每年导致1300万人死亡。在这里, 我们已经把一个小动物的血管损伤模型转化成一个大动物 (犬), 轻微的修改, 可用于临床前筛查预防和溶栓剂。除了手术方法外, 修改后的协议还描述了通过血管造影评估颈动脉疏导的分步方法, 详细说明了用大脑和颈动脉进行组织学分析以证实颈动脉疏导和脑出血, 以及具体的参数, 以完成评估下游血栓发生事件, 利用磁共振成像 (MRI)。此外, 还对以前建立良好的小动物模型进行了具体的程序性改变, 以此来转化为大动物 (犬) 血管损伤。

下面的详细程序将导致开发一个模型, 可用于预防或溶栓评估闭塞动脉干预。图 1A显示了在商业软件记录的治疗之前、期间和之后的基线流速和所产生的血流速度。该记录的数据可用于确定该犬模型中颈动脉损伤和治疗后再灌注的百分比。图 1B提供了一个例子, 对侧 (顶部) 和受伤 (底部) 犬颈动脉部分染色的 H & E, 以验证在牺牲?...

Watch this Scientific Journal Video about Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury at JoVE.com

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

在此, 我们提出了必要的修改, 以良好的特点和常用的小动物氯化铁诱导 (FeCl3) 颈动脉损伤模型, 用于大动物血管损伤模型。结果模型可用于临床前试验评估预防和溶栓的药理和机械干预。

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Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

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大鼠颈动脉球囊损伤模型试验抗血管重建治疗

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 ...

Ferric Chloride-induced Murine Thrombosis Models

三氯化铁诱导小鼠模型的血栓形成

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, ...

Invasive Hemodynamic Monitoring of Aortic and Pulmonary Artery Hemodynamics in a Large Animal Model of ARDS

ards 大型动物模型主动脉和肺动脉血流动力学的侵袭血流动力学监测

One of the leading causes of morbidity and mortality in patients with heart failure is right ventricular (RV) dysfunction, especially if it is due to ...

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 ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

大鼠胡萝卜动脉压力控制段气球损伤与腹腔ventit 治疗应用

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...