Name: comprehensive echocardiographic assessment of right ventricle function in a rat model of pulmonary arterial hypertension
Uploaded: 2023-01-20T13:45:00
Description: Watch this Scientific Journal Video about Comprehensive Echocardiographic Assessment of Right Ventricle Function in a Rat Model of Pulmonary Arterial Hypertension at JoVE.com

这项工作得到了NHLBI K01 HL155241和AHA CDA849387的支持，授予作者P.C.R.。

该协议中的所有实验均按照伊利诺伊大学芝加哥分校芝加哥机构动物护理和使用委员会的动物护理指南进行。雄性斯普拉格道利（SD）大鼠在注射MCT时体重在0.200-0.240公斤之间;但是，本文中描述的方案可用于更广泛的体重范围。这些动物是从商业来源获得的（见 材料表）。
1. 研究设计
<ol><li>动物<ol><li>获得雄性SD大鼠并让它们适应4-7天。实验组将大?...

<ol>
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F., Ryan, T., Feigenbaum, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Feigenbaum's echocardiography. 7th edn. , (2010).</li><li>Kimura, 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=Evaluation+of+right+ventricle+by+speckle+tracking+and+conventional+echocardiography+in+rats+with+right+ventricular+heart+failure.">Evaluation of right ventricle by speckle tracking and conventional echocardiography in rats with right ventricular heart failure.</a> International Heart Journal. 56 (3), 349-353 (2015).</li><li>Cheng, H. 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RV的超声心动图评估是筛选PAH动物模型中新疗法有效性的宝贵发现工具。深入表征RV结构和功能是必要的，作为治疗PAH地址RV重塑的新靶点4，14。本研究描述了一个详细的协议，可以成功表征RV的结构和功能。
复杂的结构几何形状和胸骨后面的位置使得右心室的超声心动图表征变得困难;因此，改进的超声心动图视图用于促进右心室可...

PAH 是一种进行性疾病，定义为静息时的平均肺动脉压大于 20 mmHg1。PAH 的病理改变包括肺动脉 （PA） 重塑、血管收缩、炎症以及成纤维细胞活化和增殖。这些病理变化导致肺血管阻力增加，从而导致右心室重塑、肥大和衰竭2。PAH是一种复杂的疾病，涉及几种信号通路之间的串扰。目前批准的治疗PAH的药物主要针对血管扩张剂途径，包括一氧化氮-环鸟苷单磷酸途径、前列环素途径和内皮素途径。针对这些途径的疗法已被用作单一疗法和联合疗法3，4。尽管在过去十年中PAH的治疗取得了进展，但美国REVEAL 登记处的调查结果显示，新诊断患者的 5 年生存率很低5.最近，新兴的治疗方式集中在疾病修饰剂上，这些药物可以影响PAH中发生的血管重塑的多因素病理生理学，以期破坏疾病6。
PAH的动物模型是评估新药物治疗效果的宝贵工具。MCT诱导的PAH大鼠模型是一种广泛使用的动物模型，其特征是肺动脉血管重塑，这反过来又导致肺血管阻力增加和右心室肥厚和功能障碍7，8。为了评估新疗法的疗效，研究人员通常专注于右心室压力的终末评估，而不考虑PA压力，右心室形态和右心室功能的纵向评估。使用无创和非终末成像技术对于全面检查动物模型中的疾病进展至关重要。经胸超声心动图仍然是评估动物模型中心脏形态和功能的标准方法，因为与其他成像方式（如磁共振成像）相比，经胸超声心动图成本低且易于使用。然而，由于右心室位于胸骨阴影下方、其发达的小梁和解剖形状，因此对右心室的超声心动图评估可能具有挑战性，所有这些都使得难以描绘心内膜边界9，10，11。
本文旨在描述一种综合方案，以评估 Sprague Dawley （SD） 大鼠幼稚和 MCT 诱导的 PAH 的右心室尺寸、面积和体积以及收缩和舒张功能。此外，该协议详细介绍了一种评估正常和扩张右心房超声心动图尺寸的方法。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9% sodium cloride injection USP</td>
			<td>Baxter</td>
			<td>2B1324</td>
			<td></td>
		</tr>
		<tr>
			<td>Braided cotton rolls</td>
			<td>4MD Medical Solutions</td>
			<td>RIHD201205</td>
			<td></td>
		</tr>
		<tr>
			<td>Depilating agent</td>
			<td>Wallgreens</td>
			<td>Nair Hair Remover&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode gel</td>
			<td>Parker Laboratories&#160;</td>
			<td>15-60</td>
			<td></td>
		</tr>
		<tr>
			<td>High frequency ultrasound image system and imaging station</td>
			<td>FUJIFILM VisualSonics, Inc.</td>
			<td>Vevo 2100</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>MedVet</td>
			<td>RXISO-250</td>
			<td></td>
		</tr>
		<tr>
			<td>Male sprague Dawley rats</td>
			<td>Charles River Laboratories</td>
			<td>CD 001</td>
			<td>CD IGS Rats (Crl:CD(SD))</td>
		</tr>
		<tr>
			<td>Monocrotaline (MCT)</td>
			<td>Sigma-Aldrich</td>
			<td>C2401</td>
			<td></td>
		</tr>
		<tr>
			<td>Rectal temperature probe &#160;</td>
			<td>Physitemp&#160;</td>
			<td>RET-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Sealed induction chambers</td>
			<td>Scivena Scientific</td>
			<td>RES644&#160;</td>
			<td>3 L size</td>
		</tr>
		<tr>
			<td>Solid-state array ultrasound transducer</td>
			<td>FUJIFILM VisualSonics, Inc.</td>
			<td>Vevo MicroScan transducer MS250S</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel digital calipers</td>
			<td>VWR Digital Calipers</td>
			<td>62379-531</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound gel&#160;</td>
			<td>Parker Laboratories&#160;</td>
			<td>11-08</td>
			<td></td>
		</tr>
		<tr>
			<td>Vevo Lab software</td>
			<td>FUJIFILM VisualSonics, Inc.</td>
			<td></td>
			<td>Verison 5.5.1</td>
		</tr>
	</tbody>
</table>

comprehensive echocardiographic assessment of right ventricle function in a rat model of pulmonary arterial hypertension

肺动脉高压（PAH）是一种由肺部小动脉血管收缩和重塑引起的进行性疾病。这种重塑导致肺血管阻力增加、右心室功能恶化和过早死亡。目前批准的PAH疗法主要针对肺血管扩张剂途径;然而，最近新兴的治疗方式集中在涉及疾病发病机制的其他新途径上，包括右心室（RV）重塑。允许对新疗法进行纵向评估的成像技术对于确定新药在临床前研究中的疗效非常有用。无创经胸超声心动图仍然是评估心脏功能的标准方法，并广泛用于啮齿动物模型。然而，由于其解剖位置和结构，右心室的超声心动图评估可能具有挑战性。此外，临床前啮齿动物模型中缺乏超声心动图的标准化指南，因此难以在不同实验室的研究中对RV功能进行统一的评估。在临床前研究中，大鼠久效（MCT）损伤模型被广泛用于评估治疗PAH的药物疗效。该协议描述了幼稚和MCT诱导的PAH大鼠中RV的超声心动图评估。

在这项研究中，MCT处理的大鼠被用作PAH的模型。超声心动图分析在MCT给药后的研究第23天进行，所有测量和计算均代表连续三个周期的平均值。从对照（载体：去离子水）和MCT处理的大鼠（60mg / kg）获得的超声心动图参数如 表1所示。
对照和MCT处理的大鼠的PLAX视图的代表性图像如图 1A所示。这些图像用作心脏位置和左心室形态的初步评估。R...

Watch this Scientific Journal Video about Comprehensive Echocardiographic Assessment of Right Ventricle Function in a Rat Model of Pulmonary Arterial Hypertension at JoVE.com

Comprehensive Echocardiographic Assessment of Right Ventricle Function in a Rat Model of Pulmonary Arterial Hypertension

本协议描述了肺动脉高压大鼠模型中右心室形态和功能的超声心动图特征。

肺动脉高压大鼠模型中右心室功能的综合超声心动图评估

Pulmonary arterial hypertension (PAH) is a progressive disease caused by vasoconstriction and remodeling of the small arteries in the lungs. This ...

对右心室结构和功能进行深入的超声心动图评估是筛选肺动脉高压动物模型中新型治疗靶点有效性的宝贵工具。该协议的主要优点是它提供了评估右心房和右心室形态以及表征右心室收缩和舒张功能所需的所有超声心动图视图和测量值的详细描述。该技术描述了由诺百合碱诱导的肺动脉高压大鼠模型的超声心动图特征。重要的是，超声心动图师要花足够的时间练习并精通右心室成像，即使他们以前有左心室的经验。首先在大鼠失去知觉后将大鼠从诱导室中取出，并以背褥位将它们转移到成像站动物平台上。使用连接到蒸发器的鼻锥施用异氟醚。将电极凝胶涂在每只爪子上，并将爪子固定在动物平台的心电图导板中。通过剃须胸部并使用脱毛剂去除皮毛。将直肠温度探头固定到位。将棉卷放在动物的右侧和左侧，并用胶带固定它们，以在平台倾斜时保持位置。要获取图像，请将平台向右倾斜 10 到 15 度，然后将其尾部向下约 5 度。移动换能器指向大鼠的右胸骨旁线，并降低换能器。逆时针旋转换能器，确保主动脉瓣和二尖瓣可见。将 M 模式样本体积线放在右心室最宽的区域，并调整门以包含右心室和左心室。提起换能器并重新定位，使其略微向大鼠的右胸骨旁线倾斜。将平台移动到稍微向右倾斜的位置。降低换能器，直到它与凝胶接触。将平台向右或向左移动，直到右心室流出道在视野中，肺动脉瓣聚焦且清晰可见。然后按标志存储记录肺动脉瓣的B模式图像。保持相同的 B 模式图像位置，按颜色以帮助识别通过肺动脉瓣的血流。调整速度，使最高速度点可见。按PW或脉冲波以量化血流谱。将PW角平行于通过肺动脉瓣的流动方向对齐。按更新以查看肺速度。将平台向左角倾斜，并尽可能向下倾斜。将换能器逆时针旋转 30 到 45 度，然后将换能器移动到动物的右肩或耳朵上。降低换能器，直到它与凝胶接触。确保典型的四腔视图，其中左心室和左心房可见，但胸骨阴影在右心室游离壁上。移动换能器以调整适用的四腔视图，获取右心室的聚焦视图。如果需要，稍微向尾部移动平台，直到获得最大平面。确保右心室未缩短，左心室流出道未打开。将 M 模式光标穿过右心室游离壁的三尖瓣环以可视化 TAPSE，然后按更新和签名存储以记录 TAPSE 数据。按 B 模式，然后按颜色以帮助识别通过电视的流量。按PW以量化血流谱。返回 B 模式，按压组织，并将组织多普勒样品体积门放在 RV 游离壁的三尖瓣环处。按更新以查看组织多普勒图像。使用PLAX视图，对心脏位置和左心室形态进行初步评估。使用改进的PLAX视图对右心室的评估显示，与对照大鼠相比，诺百合碱或MCT处理的大鼠右心室增大，左心室移位。在MCT处理的大鼠中观察到RVIDd，RVID和RVFWT的显着增加，表明右心室扩张和右心室游离壁增厚。修改后的PLAX视图也用于可视化肺动脉和测量肺动脉瓣直径。使用多普勒成像测量肺动脉流速表明，在对照大鼠中，肺流表现出对称的V形，在收缩中期具有峰值速度。而MCT处理的大鼠，峰值速度较慢，并且在收缩期较早发生，而在收缩后期显示缺口。采用右心室聚焦心尖四腔视图测量右心室和收缩区以及右心房面积。MCT治疗的大鼠由于肺动脉压升高而表现出右心房扩张。此外，与对照组相比，右心室压力增加导致左心室正常曲率丧失。脑室内隔也出现扁平。明亮的心室聚焦顶端四腔视图也用于测量三尖瓣环的M模式询问的TAPSE。在MCT处理的大鼠中，TAPSE显着降低，表明功能受损。该方案最具挑战性的部分是获得最佳的右心室聚焦心尖四腔视图。我们建议超声心动图师确保收缩期和舒张期整个右心室在视野范围内。使用以右心室为重点的心尖四腔视图获得的图像，以后可用于应变分析，这是一种强大的工具，可用于在初始阶段评估右心室收缩功能障碍。

Research

JoVE Journal

Biology

Laser-Induced Chronic Ocular Hypertension Model on SD Rats

激光诱导SD大鼠慢性高眼压高压模型

Glaucoma is one of the major causes of blindness in the world. Elevated intraocular pressure is a major risk factor. Laser photocoagulation induced ocular ...

科研

生物学

Right Hemihepatectomy by Suprahilar Intrahepatic Transection of the Right Hemipedicle using a Vascular Stapler

右右Hemipedicle Suprahilar肝内血管吻合器横断Hemihepatectomy

Successful hepatic resection requires profound anatomical knowledge and delicate surgical technique. Hemihepatectomies are mostly performed after ...

Long-term Silencing of Intersectin-1s in Mouse Lungs by Repeated Delivery of a Specific siRNA via Cationic Liposomes. Evaluation of Knockdown Effects by Electron Microscopy

长期沉默的1S Intersectin的在小鼠肺的特异性siRNA，通过阳离子脂质体重复交货。通过电子显微镜的击倒效果评价

Previous  studies showed that knockdown of ITSN-1s (KDITSN), an endocytic  protein involved in regulating lung vascular permeability and endothelial cells ...

A High-throughput Method for Measurement of Glomerular Filtration Rate in Conscious Mice

高通量清醒小鼠肾小球滤过率的测定方法

The measurement of  glomerular filtration rate (GFR) is the gold standard in kidney function  assessment. Currently, investigators determine GFR by ...

Phenotyping Mouse Pulmonary Function In Vivo with the Lung Diffusing Capacity

Phenotyping Mouse Pulmonary Function In Vivo with the Lung Diffusing Capacity

表型鼠标肺功能<em&gt;在体内</em&gt;与肺弥散功能

The mouse is now the primary animal used to model a variety of lung diseases. To study the mechanisms that underlie such pathologies, phenotypic methods ...

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

纯化野生型和突变CFTR蛋白的功能重建和通道活性测定

The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a unique channel-forming member of the ATP Binding Cassette (ABC) superfamily of ...

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

诱导LAP标记的稳定细胞系用于研究蛋白质功能，时空定位与蛋白质相互作用网络

Multi-protein complexes, rather than single proteins acting in isolation, often govern molecular pathways regulating cellular homeostasis. Based on this ...

A Rapid Automated Protocol for Muscle Fiber Population Analysis in Rat Muscle Cross Sections Using Myosin Heavy Chain Immunohistochemistry

在使用肌球蛋白重链免疫鼠肌肉横断面的快速自动化方案为肌纤维人口分析

Quantification of muscle fiber populations provides a deeper insight into the effects of disease, trauma, and various other influences on skeletal muscle ...

A Protocol for Using Gene Set Enrichment Analysis to Identify the Appropriate Animal Model for Translational Research

一个协议来使用基因集富集分析以确定合适的动物模型转化型研究

Recent studies that compared transcriptomic datasets of human diseases with datasets from mouse models using traditional gene-to-gene comparison ...

Implantation of Electrospun Vascular Grafts with Optimized Structure in a Rat Model

优化结构静电纺丝血管移植在大鼠模型中的植入

Here, we present a protocol to fabricate macroporous PCL vascular graft and describe an evaluation protocol by using a rat model of abdominal aorta ...