Name: a pulmonary trunk banding model of pressure overload induced right ventricular hypertrophy and failure
Uploaded: 2018-11-29T13:30:00
Description: Watch this Scientific Journal Video about A Pulmonary Trunk Banding Model of Pressure Overload Induced Right Ventricular Hypertrophy and Failure at JoVE.com

这项工作得到了丹麦独立研究理事会 [11e108410]、丹麦心脏基金会 [12e04-r90-a3852 和 12e04-r90-a3907] 和诺沃诺盘基金会 [NNF16OC0023244] 的支持。

所有大鼠都是根据丹麦动物实验法和动物实验部长令所述的丹麦国家准则进行治疗的。所有试验都得到了机构道德审查委员会的批准, 并根据丹麦动物研究法 (批准号为 2012-15-294-00384, 丹麦司法部) 进行。
1. 结扎夹的调整
请注意:肺干的条带是用一个有角度的下巴的改良的开放结扎夹贴花进行的。应用程序被修改与可调停止机制, 以停止压缩时, ?...

<ol>
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我们描述了一种方便和高度重现的方法肺躯干条带使用修改结扎夹苹果压缩肺干圈周围的钛夹。通过调整应用程序将夹子压缩到不同的内径, 可以诱导不同的 rv 肥大表型和失败, 包括严重的 rv 衰竭和心脏外失代偿表现。
虽然很简单, 但该协议包含几个关键步骤。重要的是, 老鼠在接受 ptb 手术时不能太大。根据我们的经验, 体重100–120克的 wistar 大鼠断奶者适用于该程序。在较?...

右心室 (rv) 可以适应持续的压力过载。然而, 随着时间的推移, 自适应机制无法维持心脏输出量, rv 扩张, 最终 rv 失效。rv 功能是几种心肺疾病的主要预后因素, 包括肺动脉高压 (pah)、血栓栓塞性肺动脉高压 (cteph) 和各种形式的先天性心脏病, 有压力 (或体积) 超载的 rv。尽管经过了密集的治疗, 但在这些情况下, rv 故障仍然是主要的死亡原因。
由于 rv 的独特特性1、2和胚胎发育3 , 从左心力衰竭中获得的知识不能简单地推断为右心力衰竭。因此, 右心力衰竭的动物模型是必要的, 以调查 rv 失败的机制和潜在的药理治疗策略。
有 su5416 合并缺氧 (suhx) 4 或单氯他林 (mct)5引起肺动脉高压衰竭的实验模型, 可引起肺血管疾病继发的 rv 衰竭。这些模型用于评估针对肺血管的药物的治疗效果。suhx 和 mct 模型都是 rv 失效的非固定后载模型。因此, 无法确定干预后 rv 功能的改善是否继发于后负荷减少肺血管效应, 或者是否由对 rv 的直接影响引起。此外, mct 模型有几个心脏外的影响。
在实验肺干带模型中, 由于肺干的机械收缩, 房车的后负荷是固定的。这允许调查干预对 rv 的直接心脏影响独立于任何肺血管效应6,7,8,9。通常, 条带是通过沿肺干针进行的。然后在针头和肺干周围放置一个结扎, 并用一个结捆绑, 针头被移除, 将缝合线留在肺干周围。根据针头的规格, 可以应用不同程度的收缩, 但尽管这种方法被广泛使用, 它有一些缺点。首先, 带的直径与针头的外径不完全一样, 因为结扎是绑在针和肺干上的。其次, 结的紧密程度可能会有显著的变化, 很难复制一定程度的条带。这将导致条带直径的变化, 从而更大的分散。最后, 结可能会随着时间的推移而松动。
一项研究应用了肺干10周围的半封闭钽夹。他们将肺干周围的夹子压缩到 1.10 mm 2 的内部区域,并将其与使用 18 g 针缝合的大鼠进行比较。总体而言, 与剪辑的带与较少的围手术并发症和数据差异。
基于 schou 等人11 所述的原则, 进一步发展和表征了 rv 肥大和衰竭的肺主干条带 (ptb) 模型。在这里, 我们根据以前的研究 12、13 的结果介绍我们使用这个模型的经验。对于这种模型, 钛夹被压缩在肺干周围到一个精确的预设内径, 这可以调整, 以诱导不同的 rv 失败表型。

<table border="0" cellpadding="0" cellspacing="0" width="804">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:217px;">17G IV Venflon Cannula</td>
			<td style="width:189px;">Becton Dickinson, US</td>
			<td style="width:108px;">393228</td>
			<td style="width:291px;">Distal 2 mm of the needle have been cut off</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">1 mL syringe + 26G needle&#160;</td>
			<td>Becton Dickinson, US</td>
			<td>303172 &#38; 303800</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">4-0 absorbable multifilament suture</td>
			<td>Covidien, US</td>
			<td>GL-46-MG</td>
			<td>Polysorb, violet, 5x18&#34;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">4-0 multifilament ligature</td>
			<td>Covidien, US</td>
			<td>LL-221</td>
			<td>Polysorb, violet, 98&#34;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Buprenorphine</td>
			<td>Indivior UK Limited</td>
			<td></td>
			<td>Local procurement, Temgesic 0.3 mg/mL</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Carprofene</td>
			<td>ScanVet, DK</td>
			<td>27693</td>
			<td>Norodyl 50 mg/mL</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Chlorhexidine</td>
			<td>Faaborg Pharma, DK</td>
			<td></td>
			<td>Local procurement</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Contractor</td>
			<td>Aesculap, Germany</td>
			<td>BV010R</td>
			<td>Blunt, self retaining, 70 mm</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Ear Hooklet</td>
			<td>Lawton, Germany</td>
			<td>66-0261</td>
			<td>Small, 14 cm, tip modified to an angle of 85&#176;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Eye gel</td>
			<td>Decra, UK</td>
			<td></td>
			<td>Lubrithal, Local procurement</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Forceps, Delicate Tissue&#160;</td>
			<td>Lawton, Germany</td>
			<td>09-0020</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Forceps, Dissecting&#160;</td>
			<td>Lawton, Germany</td>
			<td>09-0013</td>
			<td>1 regular, 1 with tip modified to an angle of 100&#176;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Gas Anesthesia System</td>
			<td>Penlon Limited, UK</td>
			<td>SD0217SL</td>
			<td>Sigma Delta Vaporizer</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Hair trimmer</td>
			<td>Oster</td>
			<td>76998-320-051</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Horizon Open Ligating Clip Applier</td>
			<td>Teleflex, US</td>
			<td>137085</td>
			<td>Modified with adjustable stop mechanism</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Horizon Titanium Clips</td>
			<td>Teleflex, US</td>
			<td>001200</td>
			<td>Small</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Induction chamber</td>
			<td>N/A</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Iris Scissor</td>
			<td>Lawton, Germany</td>
			<td>05-1450</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Iris Scissor&#160;</td>
			<td>Aesculap, Germany</td>
			<td>BC060R</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Mechanical ventilator</td>
			<td>Ugo Basile, Italy</td>
			<td>7025</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Microscissor</td>
			<td>Lawton, Germany</td>
			<td>63-1406</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Microscope</td>
			<td>Carl Zeiss, Germany</td>
			<td>303294-9903</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Needle Holder</td>
			<td>Lawton, Germany</td>
			<td>08-0011</td>
			<td>&#160;TITEGRIP</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Pean</td>
			<td>Lawton, Germany</td>
			<td>06-0100</td>
			<td>Halsted-Mosquito, straight</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Pro-Optha</td>
			<td>Lohmann &#38; Rauscher, Germany</td>
			<td>16515</td>
			<td>Tampon</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Saline 9 mg/mL</td>
			<td>Fresenius Kabi, DK</td>
			<td>209319</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Sevoflurane</td>
			<td>AbbVie, US</td>
			<td></td>
			<td>Sevorane, Local procurement</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Surgical hook</td>
			<td>Lawton, Germany</td>
			<td>51-0665</td>
			<td>Cushing, 19 cm, tip modified to an angle of 90&#176;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Surgical Tape</td>
			<td>3M, US</td>
			<td>1530-0</td>
			<td>Micropore</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Temperature Controller</td>
			<td>CMA Microdialysis; Sweden</td>
			<td>8003760</td>
			<td>CMA 450&#160;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Weighing machine</td>
			<td>VWR, US</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:217px;">Wistar rat weanlings</td>
			<td>Janvier Labs, France</td>
			<td style="width:108px;"></td>
			<td>RjHan:WI, 100-120 g</td>
		</tr>
	</tbody>
</table>

a pulmonary trunk banding model of pressure overload induced right ventricular hypertrophy and failure

持续压力过载引起的右心室衰竭是导致几种心肺疾病发病和死亡的主要原因。因此, 有必要建立可靠和可重复的 rv 故障动物模型, 以调查疾病机制和潜在治疗策略的效果。肺干带是一种常见的方法, 以诱导孤立的 rv 肥大, 但一般来说, 前面描述的模型没有成功地创建一个稳定的模型的 rv 肥大和失败。
我们提出了一个大鼠模型的压力超载诱导 rv 肥大引起的肺干带 (ptb), 使不同的表型的 rv 肥大与 rv 失败和没有。我们使用改良的结扎夹贴花将肺主干周围的钛夹压缩到预先设定的内径。我们使用不同的剪辑直径, 以诱导不同阶段的疾病进展, 从轻度 rv 肥大到失代偿 rv 失败。
rv 肥大在接受 ptb 手术的大鼠中持续发展, 根据应用带夹的直径, 我们可以准确地再现不同的疾病严重程度, 从补偿肥大到严重的失代偿性 rv 不等心脏外表现的失败。
所提出的 ptb 模型是压力过载诱导 rv 肥大和失效的有效而稳健的模型, 它对其他带状模型具有多大的优点, 包括高重现性和诱发严重和失代偿的 rv 故障的可能性。

使用我们第12、13 组先前研究中描述的 ptb 程序, 我们通过带 1.0 mm 夹、中等程度的 rv 失效 (ptb) 和 0.6 mm 夹的带状和严重程度来诱发 rv 肥大 (ptb 轻度)rv 故障 (ptb 严重) 的带与0.5 毫米剪辑。受严重条带的大鼠出现心脏外 rv 衰竭的表现, 包括肝功能衰竭和腹水 (图 5e)。所有大鼠在 ptb 后7周后都接受了评估, 假老?...

Watch this Scientific Journal Video about A Pulmonary Trunk Banding Model of Pressure Overload Induced Right Ventricular Hypertrophy and Failure at JoVE.com

A Pulmonary Trunk Banding Model of Pressure Overload Induced Right Ventricular Hypertrophy and Failure

我们提出了一种手术方法, 以诱导右心室肥厚和失败的大鼠。

压力超载诱发右室肥厚和衰竭的肺动脉扣条柱模型

Right ventricular (RV) failure induced by sustained pressure overload is a major contributor to morbidity and mortality in several cardiopulmonary ...

此方法可以帮助回答有关基础机制和右心室故障处理的关键问题。这种技术的主要优点是，它允许一个精确和可重复的后载量增加，诱导右心室肥大和故障。该技术的影响延伸到右心室衰竭的治疗，因为它消除了对肺血管扩张的次要影响。在开始主干隔离程序之前，为带状选择适当的直径，并调整连杆夹贴动器，直到夹口完全压缩时钳口之间的距离为 1 毫米，留下 6 毫米的流明。接下来，确认在麻醉的100至120克威斯塔大鼠断奶时，在手指趾捏中缺乏反应，并沿着胸骨的中线进行两厘米的皮肤切口。在解剖显微镜下切割主要胸肌的胸腔附着，并识别第二个、第三和第四个骨骼。使用一把剪刀在胸骨附近切割第四、第三和第二个骨骼，并仔细解剖骨骼间肌肉，直到完成完整的左胸腔切除术。当进行左横向胸腔切除术时，你应该非常小心，因为左内动脉只是波纹管，所以要非常小心。在胸骨和骨子之间插入缩回器，然后打开钳口，直到胸腺在观察场的顶部可见。使用钳子小心地翻转胸腺，露出主动脉和肺干，并引导一个小手术钩的尖端，弯曲到85度角，通过左主试附属物后面的反向左心鼻窦。将吊钩通过鼻窦中拉半回，并引导耳钩的尖端向上，直到它出现在上升主动脉和肺躯干之间。放置耳钩时，请确保角度正确。你不应该使用任何力量， 它最初可能有点困难， 但你可以通过练习掌握它。使用虹膜剪刀去除任何覆盖尖端的结缔组织，将肺干从上升主动脉分离。使用倾斜的前导引导 10 厘米的 4/0 多光连接字穿过肺干干，穿过钩的通道。用另一个钳子抓住连字的末尾。肺干干现在应该从上升主动脉分离，并可以通过它周围的连字控制。当肺干干已固定时，使用夹子加载调整的连体夹贴画。小心地引导夹子的一个钳口和一条腿穿过肺躯干周围的通道，使用连体字轻轻地将肺躯干向上拉入夹子的叉子。当肺干箱在叉子中，并且夹子的两个尖端没有任何结缔组织时，压缩夹子以在右心室扩张以响应带状时应用带状去除连字。当树干被绑带时，从胸腺中取出豌豆钳，将组织返回到其自然位置。拆下缩回器，并使用可吸收的 4/0 多丝缝合线将胸部合三层。然后下皮注射2毫升盐水，以取代手术过程中丢失的任何液体，并监测动物，直到完全恢复。选择夹子的不同直径，可以诱发疾病严重程度的不同阶段，这一点通过右心室肥大程度的增加就可见一下。只有严重的带状导致额外的心脏表现，包括肝衰竭。中度和重肺主带的差异通过增加右心室压力而明显，并导致中度与重重右心室衰竭，如心脏输出的逐步减少和三叶花环形平面收缩偏移随着带状严重性的增加而显现出来。肺躯干带状手术诱导右心室扩张，明显表现为中度肺干带大鼠右心室端舒张体积和收缩体积的增加，与假操作动物相比，在重度肺躯干带状大鼠中，与中度和假动物相比。也观察到右心室喷射分数的逐步减少。右心室肥大进一步表明，与假操作的动物相比，肺躯干带大鼠的心肌细胞横截面面积增加，以及右心室衰竭（包括右心室纤维化）的右心室的其他形态变化。一旦掌握了这项技术，如果执行得当，可以在30分钟内执行。在尝试此过程时，在开始实验研究之前，获得与模型一致的结果非常重要。这项技术开发后，为右心力衰竭领域的研究人员探索药物干预和基本机制铺平了道路。看完这个视频后，你应该有一个很好的理解如何执行肺中继带。

Research

JoVE Journal

Medicine

Echocardiographic Assessment of the Right Heart in Mice

右心小鼠的超声心动图评价

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

科研

医学

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

大鼠建立压力超负荷心肌肥厚模型在升主动脉缩窄

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

右心室结构和功能的肺动脉缩窄经胸超声小鼠模型的评估

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

Chronic Thromboembolic Pulmonary Hypertension and Assessment of Right Ventricular Function in the Piglet

慢性血栓栓塞性肺动脉高压和右心室功能的仔猪评估

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. ...

Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension

分流手术，右心导管，血管形态计量学在大鼠模型中的流量引起的肺动脉高压

In this protocol, PAH is induced by combining a 60 mg/kg monocrotalin (MCT) injection with increased pulmonary blood flow through an aorto-caval shunt ...

Tachycardia-Induced Cardiomyopathy As a Chronic Heart Failure Model in Swine

心动过速诱发的心肌病作为猪慢性心力衰竭模型的研究

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment ...

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse

小鼠右心室舒张参数的超声心电图测量

Diastolic dysfunction is a prominent feature of right ventricular (RV) remodeling associated with conditions of pressure overload. However, the RV ...

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

慢性血栓栓塞性肺动脉高压大型动物模型中急性右心衰竭的诱导和表型分析

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

小鼠右心室系统及缺氧诱发肺动脉高血压的侵入性血液动力学评估

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...

A Rat Model of Pressure Overload Induced Moderate Remodeling and Systolic Dysfunction as Opposed to Overt Systolic Heart Failure

压力过载诱导中度重塑和收缩功能障碍的大鼠模型，而不是过度收缩性心力衰竭

In response to an injury, such as myocardial infarction, prolonged hypertension or a cardiotoxic agent, the heart initially adapts through the activation ...