Name: measurement of pulse propagation velocity, distensibility and strain in an abdominal aortic aneurysm mouse model
Uploaded: 2020-02-23T13:00:00
Description: Watch this Scientific Journal Video about Measurement of Pulse Propagation Velocity, Distensibility and Strain in an Abdominal Aortic Aneurysm Mouse Model at JoVE.com

这项工作得到了R01HL124155（CPH）的支持，以及密苏里大学研究所对CPH的资助。

处理小鼠和超声成像的规程得到了密苏里大学动物护理和使用委员会的批准（动物协议号8799），并根据AAALAC国际进行。
1. 小鼠的设备设置和制备
<ol><li>设备设置<ol>
<li>打开超声波仪器、超声波凝胶加热器和加热垫。</li>
		<li>打开超声波程序，输入每个鼠标的研究名称和描述性信息。</li>
		<li>选择应用程序作为常规映像。</li>
		<li>选择合适的传感器进行腹部成像...

<ol>
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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Notch+signalling:+a+simple+pathway+becomes+complex.">Notch signalling: a simple pathway becomes complex.</a> Nature Reviews Molecular and Cell Biology. 7, 678-689 (2006).</li><li>Hans, C. P., 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=Inhibition+of+Notch1+signaling+reduces+abdominal+aortic+aneurysm+in+mice+by+attenuating+macrophage-mediated+inflammation.">Inhibition of Notch1 signaling reduces abdominal aortic aneurysm in mice by attenuating macrophage-mediated inflammation.</a> Arteriosclerosis, Thrombosis and Vascular Biology. 32, 3012-3023 (2012).</li><li>Cheng, J., Koenig, S. N., Kuivaniemi, H. S., Garg, V., Hans, C. 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I., 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=Evaluation+of+aortic+stiffness+(aortic+pulse-wave+velocity)+before+and+after+elective+abdominal+aortic+aneurysm+repair+procedures:+a+pilot+study.">Evaluation of aortic stiffness (aortic pulse-wave velocity) before and after elective abdominal aortic aneurysm repair procedures: a pilot study.</a> Open Cardiovascular Medicine Journal. 3, 173-175 (2009).</li><li>Fortier, C., Desjardins, M. P., Agharazii, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aortic-Brachial+Pulse+Wave+Velocity+Ratio:+A+Measure+of+Arterial+Stiffness+Gradient+Not+Affected+by+Mean+Arterial+Pressure.">Aortic-Brachial Pulse Wave Velocity Ratio: A Measure of Arterial Stiffness Gradient Not Affected by Mean Arterial Pressure.</a> Pulse. 5, 117-124 (2017).</li><li>Golledge, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abdominal+aortic+aneurysm:+update+on+pathogenesis+and+medical+treatments.">Abdominal aortic aneurysm: update on pathogenesis and medical treatments.</a> Nature Reviews Cardiology. 16 (4), 225-242 (2019).</li><li>Choksy, S. A., Wilmink, A. B., Quick, C. 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超声成像提供了一种强大的技术，通过测量PPV、不透度和径向应变来确定主主的功能特性。这些测量对于研究AAA的小鼠模型特别有指导意义，并且体内方法允许收集对了解主动脉病理学的时间发展可能很重要的纵向数据。具体来说，在腹部主动脉的测量通过PPV、不均匀性和径向应变在腹部主动脉中通过分析EKV数据确定，并被认为是AAA不稳定性16的独立危险因素。这些协议中描述?...

腹部主动脉瘤（AAA）代表一种严重的心血管疾病，其特征是主动脉的永久局部扩张，超过原始血管直径的1.5倍1。AAA在美国死亡率排名前13位。AAA的进展归因于主动脉壁的退化和乳突分裂，最终导致主动脉破裂。主动脉壁的这些变化可能发生，没有显著增加最大宫内直径（MILD），从而表明，仅MILD不足以预测疾病的严重程度3。因此，需要确定其他因素来检测主动脉壁的初始变化，这可能指导早期治疗方案。该协议的总体目标是为使用超声波成像（以脉冲传播速度 （PPV）、不扩展性和径向应变的测量为特征，为评估主动脉功能特性提供实用指南。
Daugherty及其同事首先描述的一种研究AAA的具有良好特征的实验模型，涉及通过Apoe-/-小鼠4的渗透泵进行血管紧张症II（AngII）的皮下输注。使用超声波成像对MILD的精确测量有助于描述这种小鼠模型5中的AAA。虽然在AAA开发过程中组织学的变化得到了广泛的研究，但船壁的功能特性变化，如主动脉刚度，并没有得到很好的描述。该协议强调将高频超声与精密分析相结合作为研究AAA时间进展的有力工具。具体来说，这些方法使我们能够评估由PPV、不强度和径向应变测量的容器壁的功能特性。
最近对AAA的人类受试者以及鼠乳酸酶诱导的AAA模型的临床研究表明，主动脉刚度与主动脉直径6、7之间存在正相关关系。PPV是主动脉刚度的指标，被认为是量化血管壁6、8刚度变化的极佳测量指标。PPV是通过测量脉冲波形在血管沿线两个位点的传递时间来计算的，从而提供主动脉刚度的区域评估。我们最近已经证明，PPV测量的主动脉刚度增加，在使用原子力显微镜确定的细胞水平上，与动脉瘤发育正相关9。此外，文献还表明，主动脉刚度可能先于动脉扩张，因此在AAA10开发过程中，可提供有关血管壁区域内在特性的有用信息。同样，不测量性和应变测量是测量动脉适应性早期变化的定量工具。健康的动脉是灵活和弹性的，而随着刚度增加和弹性降低，不粘度和应变降低。在这里，我们为使用高频超声系统测量小鼠的MILD、PPV、失散性和径向应变提供了实用的指南和分步协议。该协议提供了技术方法，应结合特定超声成像仪器手册和随附的视频教程提供的基本信息进行使用。重要的是，在我们手中，所述的成像协议提供了可重复的准确数据，这些数据在实验AAA的发展和进展研究中显得有价值。
为了进一步证明超声成像的效用，我们提供了从我们自己的研究中拍摄的示例图像和测量，这些研究旨在使用药理学方法来预防实验性AAA11。具体来说，切口信号已被建议涉及血管发育和炎症的多个方面12。使用基因单倍素和药理学方法，我们先前已经证明，Notch抑制通过防止巨噬细胞在血管损伤部位的渗透，减少小鼠AAA的发展。对于目前的文章，使用药理方法的Notch抑制，我们专注于主动脉刚度和与AAA相关的因素之间的关系。这些研究表明，Notch抑制降低主动脉刚度，这是AAA级11的量度。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Angiotensin II</td>
			<td>Sigma</td>
			<td>A9525</td>
			<td></td>
		</tr>
		<tr>
			<td>Apoe-/- mice</td>
			<td>The Jackon lab</td>
			<td><img src="/files/ftp_upload/60515/60515mateq.jpg"/></td>
			<td></td>
		</tr>
		<tr>
			<td>Clippers</td>
			<td>WAHL</td>
			<td>1854</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swab</td>
			<td>Q-tips</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DAPT</td>
			<td>Sigma</td>
			<td>D5942</td>
			<td></td>
		</tr>
		<tr>
			<td>Depilatory cream</td>
			<td>Nair</td>
			<td>LL9038</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode cream</td>
			<td>Sigma</td>
			<td>17-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Gel warmer</td>
			<td>Thermasonic (Parker)</td>
			<td>82-03 (LED)</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Stryker</td>
			<td>T/pump professional</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>VetOne</td>
			<td>Fluriso TM</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane vaporizer</td>
			<td>Visualsonics</td>
			<td>VS4244</td>
			<td></td>
		</tr>
		<tr>
			<td>Lubricating ophthalmic ointment</td>
			<td>Lacri-lube</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Osmotic pumps</td>
			<td>Alzet</td>
			<td>Model 2004</td>
			<td></td>
		</tr>
		<tr>
			<td>Oxygen tank</td>
			<td>Air gas</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tranducer</td>
			<td>Visualsonics</td>
			<td>MS-400 or MS550D</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic gel</td>
			<td>Parker</td>
			<td>Aquasonic clear</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound Imaging System</td>
			<td>Visualsonics</td>
			<td>Vevo 2100</td>
			<td></td>
		</tr>
		<tr>
			<td>Vevo Vasc Software</td>
			<td>Visualsonics</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

measurement of pulse propagation velocity, distensibility and strain in an abdominal aortic aneurysm mouse model

腹部主动脉瘤 （AAA） 定义为腹部主动脉的局部扩张，其内光直径 （MILD） 超过其原始尺寸的 1.5 倍。临床和实验研究表明，小动脉瘤可能破裂，而大动脉瘤的亚群可能保持稳定。因此，除了测量主肠的内光直径外，对船壁结构特征的了解可以为评估AAA的稳定性提供重要信息。主动脉硬化最近成为确定血管壁早期变化的可靠工具。脉冲传播速度 （PPV） 以及不透度和径向应变是用于评估主动脉刚度的非常有用的超声方法。该协议的主要目的是提供一种全面的技术，用于使用超声成像系统获取图像并分析由MILD、PPV、不透度和径向应变确定的主主的结构和功能特性。

图2A和图2B分别显示了小鼠正常和动脉瘤主动脉的代表性M模式图像。 上腹主动脉由右肾动脉和上流的动脉旁的位置确定（图2A）。在收缩性心脏周期的三个不同心跳处，在正常动脉瘤和动脉瘤中，用于计算MILD的代表性图像分别如图2C，D所示。在主动脉瘤已?...

Watch this Scientific Journal Video about Measurement of Pulse Propagation Velocity, Distensibility and Strain in an Abdominal Aortic Aneurysm Mouse Model at JoVE.com

Measurement of Pulse Propagation Velocity, Distensibility and Strain in an Abdominal Aortic Aneurysm Mouse Model

本手稿描述了使用高频超声成像测量腹部主动脉瘤小鼠模型上的光径、脉冲传播速度、不强度和径向应变的详细方案。

腹部主动脉瘤小鼠模型中脉冲传播速度、失散性和应变的测量

An abdominal aortic aneurysm (AAA) is defined as a localized dilation of the abdominal aorta that exceeds the maximal intraluminal diameter (MILD) by 1.5 ...

超声成像和功能分析是非常强大的技术，以确定腹部主动脉瘤的生长和稳定性，超越传统的主动脉大小和直径的方法。这种非侵入性技术测量脉冲波速度、距离能力和大音距离，以监测大音库功能与大音直径的细微变化。主动脉僵硬是腹部主动脉瘤进展的指标，稳定性可用于评估临床前试验中感兴趣的治疗剂的效果。此外，速度和应变测量也可以与各种心血管疾病（如高血压、糖尿病、肥胖症和动脉硬化）发展过程中发生的血管变化相关。演示该程序将是来自成像核心设施的研究助理教授Sun博士。为使小鼠为成像程序做好准备，在确认踏板反射反应不足后，对麻醉小鼠的眼睛应用视点溶液，将鼠标放在动物成像平台上右下倾斜 10 度的超前位置。将电极凝胶涂抹到所有四爪上，将爪子胶带涂抹在成像平台上的铜心电图引线上。使用剪子在成像现场剃毛，并在暴露的皮肤上涂抹脱毛霜。不到一分钟后，使用湿纸巾去除奶油和头发，并应用37摄氏度的超声波凝胶到准备的场地。然后使用棉签的木端来平滑凝胶并去除任何气泡。对于腹部主动脉成像，将传感器连接到其支架，并放置垂直于鼠标中线的传感器。降低传感器，直到它接触凝胶，将尖端定位在隔膜下方 0.5 至 1 厘米。当传感器就位时，在短轴视图中可视化腹部主动脉。接下来，顺时针轻轻旋转传感器 90 度，然后缓慢调整 X 轴微操纵器旋钮，在长轴视图中可视化大奥。使用焦点区域和焦点深度切换来设置大项区域的焦点区域和深度。手动调整时间增益补偿滑块，使大项的流明变暗，以实现大项墙的最佳对比度。调整 Y 轴操纵器以可视化上部肠动脉和右肾动脉的分支点。正确的肾动脉可以用作捕获超级肾主动脉图像的地标。在超级肾大图上录制至少 100 帧 B 模式图像后，保存图像并选择结束模式。滚动光标，将黄色指示线带到具有清晰容器壁图像的正常大动脉部分或观察到动脉瘤最大直径的部分。单击 SV 门切换并调整光标，以确保容器壁包含在测量支架中。然后按更新记录 M 模式测量值，然后发送到存储以捕获测量值。要获取 EKV 图像，请按 B 模式按钮返回 B 模式录制，然后按"物理设置"按钮以允许选择呼吸门控。手动调整浇注延迟和窗口，仅在呼吸波的扁平部分记录数据。记录部分将显示为追踪呼吸波的彩色块。按 EKV 按钮以启用 EKV 模式。在相应的菜单中，将采集类型设置为标准分辨率，将帧速率设置为 3，000 或更高，然后单击"扫描"以录制 EKV 图像。然后按发送到存储以保存图像，并使用 EKV 模式图像获取 PPV、可分散性和径向应变的测量值。要测量轻度，请打开超声波程序，并输入每只小鼠的研究名称和描述性信息。打开超声波数据和 M 模式图像并暂停心跳。单击测量值，然后从下拉选项中选择血管包和深度。然后绘制一条线穿过从内墙延伸到墙壁的大音流明。对于 PPV 分析，打开 EKV 图像并暂停检测信号。单击该图标可打开分析软件上的新窗口，然后选择 PPV 选项。将出现一个小窗口，其图像是大项。单击上容器壁并拖动指针，用大约四毫米框覆盖超级肾大母的两面墙，然后单击"接受"以保存 PPV 值。要在打开的 EKV 图像中进行可分散性分析，请单击软件图标并选择"跟踪感兴趣的新区域"，在容器的两面图上绘制一个矩形框。该软件将自动跟踪容器的上壁和下壁。如有必要，单击绿点以调整轨迹对齐，然后单击"接受"。该软件将计算所选感兴趣区域内的可扩展性。要获得径向应变测量，请从菜单中选择适当的应变选项，然后单击应变。径向应变和切向应变的图像将打开。然后将光标移动到曲线的峰值，以获取径向应变的值。为了获得最佳的可重复性和某些阶段的检测，图像应由同一人在同一解剖位置进行分析，以确保数据中的一致性。在这里，显示了正常和动脉瘤小鼠腹部主动脉的代表性M模式图像。超级肾腹主动脉可以通过其位置识别肾动脉的右分支和优越的肠动脉。如这些图像所示，在正常和动脉瘤中，可以使用收缩性心脏周期的三种不同的心跳来计算轻度。在动脉瘤的情况下，发光直径可以通过在最大扩张区域的流明的两个内边缘之间绘制一条垂直的黄线来确定。在这项具有代表性的分析中，跨腹部超声成像显示，轻度和PPV的逐渐增加，以及第28天对血管紧张素II的反应的可分散性和径向应变的减少。血管紧张素II输液从第28天到56日略有增加，而使用一种缺口抑制剂的治疗与血管他汀II相比没有显著影响轻度。值得注意的是，PPV与第28天温和密切相关，而在第56天，相关性相对较弱。组织学上，血管紧张素II输液增加了大干的介质层的胶原蛋白降解和蛋白解活性。然而，槽抑制剂治疗最大限度地减少了细胞基质外降解的这些变化。真正设置动物和探头和识别腹部主动脉是最关键的步骤。初学者我建议使用微操纵旋钮进行微调。使用这种方法，我们还可以测量主动脉或肾动脉的块流，评估心脏功能，并监测周围血管疾病。达到一定尺寸后，腹主动脉瘤不会过度生长。在这些情况下，这些技术会迅速检测出某些结构变化，以响应药理剂或其他操作。在麻醉时，请务必使用木炭和溶剂进行异氟和排气磨损。还要确保鼻孔没有泄漏。

Research

JoVE Journal

Immunology and Infection

Colon Ascendens Stent Peritonitis (CASP) - a Standardized Model for Polymicrobial Abdominal Sepsis

Colon Ascendens Stent Peritonitis CASP - a Standardized Model for Polymicrobial Abdominal Sepsis

科隆Ascendens支架腹膜炎（CASP） - 标准化的模式为多种微生物腹部败血症

Sepsis remains a persistent problem on intensive care units all over the world. Understanding the complex mechanisms of sepsis is the precondition for ...

科研

免疫与感染

Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis

细菌性前列腺炎小鼠模型的测量触觉异常

Uropathogenic Escherichia coli (UPEC) are pathogens that play an important role in urinary tract infections and bacterial prostatitis1. We have recently ...

Influenza Virus Propagation in Embryonated Chicken Eggs

在鸡胚的流感病毒传播

Influenza infection is associated with about 36,000 deaths and more than 200,000 hospitalizations every year in the United States. The continuous ...

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

高通量设置的建立筛选小分子能调节C-二GMP信令<em&gt;绿脓杆菌</em

Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. ...

Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291

Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291

头孢哌酮治疗小鼠模型的临床相关的<em&gt;艰难梭菌</em&gt;应变R20291

Clostridium difficile is an anaerobic, gram-positive, spore-forming enteric pathogen that is associated with increasing morbidity and mortality and ...

Effects of Taste Signaling Protein Abolishment on Gut Inflammation in an Inflammatory Bowel Disease Mouse Model

味觉信号蛋白废除对炎症性肠病小鼠肠道炎症的影响

Inflammatory bowel disease (IBD) is one of the immune-related gastrointestinal disorders, including ulcerative colitis and Crohn's disease, that affects ...

Experimental Autoimmune Uveitis: An Intraocular Inflammatory Mouse Model

实验性自身免疫性葡萄膜炎：眼内炎症小鼠模型

Experimental Autoimmune Uveitis (EAU) is driven by immune cells responding to self-antigens. Many features of this non-infectious, intraocular ...

Therapeutic Evaluation of Fecal Microbiota Transplantation in an Interleukin 10-Deficient Mouse Model

With the development of microecology in recent years, the relationship between intestinal bacteria and inflammatory bowel disease (IBD) has attracted ...

A Mouse Model of Chronic Liver Fibrosis for the Study of Biliary Atresia

慢性肝纤维化小鼠模型用于研究胆道闭锁

Biliary atresia (BA) is a fatal disease involving obstructive jaundice, and it is the most common indication for liver transplantation in children. Due to ...

Single&#45;Copy Gene Expression for Characterizing Multidrug Efflux Systems

Characterizing Multidrug Efflux Systems in Acinetobacter baumannii Using an Efflux&#45;Deficient Bacterial Strain and a Single&#45;Copy Gene Expression System

Author Spotlight: Advancing Antibiotic Resistance Research Using an Efflux&#45;Deficient Bacterial Strain and a Single&#45;Copy Gene Expression System

Acinetobacter baumannii is recognized as a challenging Gram-negative pathogen due to its widespread resistance to antibiotics. It is crucial to comprehend ...