Name: studying left ventricular reverse remodeling by aortic debanding in rodents
Uploaded: 2021-07-14T13:00:00
Description: Watch this Scientific Journal Video about Studying Left Ventricular Reverse Remodeling by Aortic Debanding in Rodents at JoVE.com

作者感谢葡萄牙科学和技术基金会（FCT）、欧洲联盟、国家科学院（QREN）、欧洲德森沃尔维门托地区基金会（FEDER）和竞争因素方案（竞争）为联合IC（UID/IC/00051/2013）研究单位提供资金。该项目由联邦贸易委员会通过竞争2020 - 国际竞争项目（POCI）支持，项目DOCNET（NORTE-0145-FEDER-0000003），由北葡萄牙区域业务方案（NORTE 2020）根据葡萄牙2020年伙伴关系协定支持， 通过欧洲区域发展基金（ERDF），该项目NETDIAMOND（POCI-01-0145-FEDER-016385），由欧洲结构和投资基金支持，里斯本2020年区域业务计划。丹妮拉·米兰达-西尔瓦和帕特里西亚·罗德里格斯分别由特克诺洛尼亚基金会（FCT）通过奖学金赠款（SFRH/BD/87556/2012和SFRH/BD/96026/2013）提供资金。

所有动物实验均符合《实验室动物护理和使用指南》（NIH出版物第85-23号，2011年修订）和葡萄牙动物福利法（DL 129/92，DL 197/96;P 1131/97）。地方当局主管部门批准了这项实验议定书（018833）。七周大的雄性C57B1/J6小鼠被关在适当的笼子里，有规律的12/12 h光暗循环环境，温度为22°C，湿度为60%，可获得水和标准饮食。
1. 手术场的准备
<ol><li>用70%的酒精对手术部位进行消毒，并在...

<ol>
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本文提出的模型分别模拟了主动脉绑带和解散后LV改造和RR的过程。因此，它代表了一个很好的实验模型，以推进我们对与不利LV改造有关的分子机制的知识，并测试新的治疗策略，能够诱导这些患者的心肌恢复。该协议详细说明了如何创建主动脉带和脱体的啮齿动物模型的步骤，使用微创和高度保守的手术技术，以减少手术创伤。
协议最关键的步骤与主动脉带期间的手术攻?...

小鼠横向或上升主动脉的收缩是压力过载引起的心脏肥大、舒张和收缩功能障碍以及心力衰竭1、2、3、4的广泛应用的实验模型。主动脉收缩最初导致补偿左心室（LV）同心肥大，使壁应力正常化1。然而，在某些情况下，如长时间心脏超负荷，这种肥大不足以减少壁应力，触发舒张和收缩功能障碍（病理肥大）5。同时，细胞外基质 （ECM） 的变化导致胶原蛋白沉积和交叉链接的过程称为纤维化，可以细分为替代纤维化和反应性纤维化。纤维化是，在大多数情况下，不可逆转和妥协心肌恢复后超载救济6，7。然而，最近的心脏磁共振成像研究表明，反应性纤维化能够长期倒退。总之，纤维化、肥大和心脏功能障碍是心肌重塑过程的一部分，这个过程会迅速走向心力衰竭（HF）。
了解心肌重塑的特点已成为限制或扭转其进展的主要目标，后者称为反向改造 （RR）。RR 一词包括任何由给定干预长期逆转的心肌改变，如药理疗法（如抗高血压药物）、瓣膜手术（如主动脉狭窄）或心室辅助设备（如慢性 HF）。然而，由于普遍存在的肥大或收缩/舒张功能障碍，RR通常不完整。因此，RR 基本机制和新颖的治疗策略的澄清仍然缺失，这主要是因为大多数患者在 RR 期间无法访问和研究人类心肌组织。
为了克服这一限制，啮齿动物模型在促进我们对高频进展中涉及的信号通路的理解方面发挥了重要作用。具体来说，主动脉收缩小鼠的主动脉去除是研究不利LV改造9和RR10，11背后的分子机制的有用模型，因为它允许在这两个阶段的不同时间点收集心肌样本。此外，它提供了一个优秀的实验设置，以测试潜在的新目标，可以促进/加速RR。例如，在主动脉狭窄的背景下，该模型可能提供有关 RR6、12完整性基础下心肌反应的巨大多样性所涉及的分子机制的信息，以及瓣膜更换的最佳时机，这是当前知识的一个主要缺陷。事实上，这种干预的最佳时机是一个争论的话题，主要是因为它是根据主动脉梯度大小来定义的。一些研究主张，这个时间点可能为时已晚，心肌恢复，因为纤维化和舒张功能障碍往往已经存在12。
据我们所知，这是唯一的动物模型，回顾心肌改造和RR的过程分别发生在主动脉狭窄或高血压等条件下的阀门更换或抗高血压药物的开始。
为了应对上述挑战，我们描述了一种可在小鼠和大鼠身上实施的手术动物模型，解决了这两个物种之间的差异。我们描述了进行这些手术时涉及的主要步骤和细节。最后，我们报告LV在RR之前和整个RR中发生的最重大变化。

<table border="0" cellpadding="0" cellspacing="0" style="width:965px;" width="964">
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	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Absorption Spears</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">18105-03</td>
			<td style="width:393px;">To absorb fluids during the surgery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Blades</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">10011-00</td>
			<td>To perform the skin incision</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenorphine</td>
			<td style="width:145px;">Buprelieve</td>
			<td style="width:161px;"></td>
			<td>Analgesia drug</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Catutery</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">18010-00</td>
			<td style="width:393px;">To prevent exsanguination</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Catutery tips</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">18010-01</td>
			<td style="width:393px;">To prevent exsanguination</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">cotton swab</td>
			<td style="width:145px;">Johnson&#39;s</td>
			<td style="width:161px;"></td>
			<td style="width:393px;">To absorb fluids during the surgery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Depilatory cream</td>
			<td style="width:145px;">Veet</td>
			<td style="width:161px;"></td>
			<td>To delipate the animal</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:18px;">Disposable operating room table cover</td>
			<td style="width:145px;">MEDKINE</td>
			<td style="width:161px;">DYND4030SB</td>
			<td>To cover the surgical area</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Echo probe</td>
			<td style="width:145px;">Siemens</td>
			<td>Sequoia 15L8W</td>
			<td>Ultrasound signal aquisition</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Echocardiograph</td>
			<td style="width:145px;">Siemens</td>
			<td>Acuson Sequoia C512</td>
			<td>Ultrasound signal aquisition</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:265px;">End-tidal CO2 monitor</td>
			<td style="width:145px;">Kent Scientific</td>
			<td style="width:161px;">CapnoStat</td>
			<td>To control expiration gas saturation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Forcep/Tweezers</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">11255-20</td>
			<td>To dissect the tissues and aorta</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Forcep/Tweezers</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">11272-30</td>
			<td>To dissect the tissues and aorta</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Forcep/Tweezers</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">11151-10</td>
			<td>To dissect the tissues and aorta</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Forcep/Tweezers</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">11152-10</td>
			<td>To dissect the tissues and aorta</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Gas system</td>
			<td>Penlon Sigma Delta</td>
			<td style="width:161px;"></td>
			<td>To anesthesia and mechanical ventilation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Hemostats</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">13010-12</td>
			<td>To hold the suture before tight the aorta</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Hemostats</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">13011-12</td>
			<td>To hold the suture before tight the aorta</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Ligation aids</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">18062-12</td>
			<td>To place a suture around the aorta</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Magnetic retractor</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">18200-20</td>
			<td>To help keep the animal in a proper position</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Needle holder</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">12503-15</td>
			<td>To suture the animal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Needle 26G</td>
			<td style="width:145px;">B-BRAUN</td>
			<td style="width:161px;">4665457</td>
			<td>To serve as a molde of aortic constriction diameter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Oxygen</td>
			<td style="width:145px;">Air Liquide</td>
			<td style="width:161px;"></td>
			<td>To anesthesia and mechanical ventilation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Polipropilene suture</td>
			<td style="width:145px;">Vycril</td>
			<td style="width:161px;">W8304/W8597</td>
			<td>To suture the animal and to do the constriction</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Povidone-iodine solution</td>
			<td style="width:145px;">Betadine&#174;</td>
			<td style="width:161px;"></td>
			<td>Skin antiseptic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">PowerLab</td>
			<td>Millar instruments</td>
			<td>ML880 PowerLab 16/30</td>
			<td>PV loop Signal Aquisition</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Pulse oximeter</td>
			<td style="width:145px;">Kent Scientific</td>
			<td style="width:161px;">MouseStat</td>
			<td>To control heart rate and blood saturation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">PVAN software</td>
			<td style="width:145px;">Millar Instruments</td>
			<td style="width:161px;"></td>
			<td>To analyse the haemodynamic data</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">PV loop cathether</td>
			<td>Millar instruments</td>
			<td>SPR-1035. 1.4 F</td>
			<td>PV loop Signal Aquisition</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Retractor</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">17000-01</td>
			<td>To provide a better overview of the aorta</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Scalpet handle</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">10003-12</td>
			<td>To perform the skin incision</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Scissors</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">15070-08</td>
			<td>To cut the suture in debanding surgery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Scissors</td>
			<td style="width:145px;">F.S.T</td>
			<td style="width:161px;">14084-09</td>
			<td>To cut other material during the surgery e.g. suture, papper</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Sevoflurane</td>
			<td style="width:145px;">Baxter</td>
			<td style="width:161px;">533-CA2L9117</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Temperature control module</td>
			<td style="width:145px;">Kent Scientific</td>
			<td style="width:161px;">RightTemp</td>
			<td>To control animal corporal temperature</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Ventilator</td>
			<td style="width:145px;">Kent Scientific</td>
			<td style="width:161px;">PhysioSuite</td>
			<td>To ventilate the animal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Water-bath</td>
			<td style="width:145px;">Thermo Scientific&#8482;</td>
			<td style="width:161px;">TSGP02</td>
			<td>To maintain water temperature adequate to heat the P-V loop catethers</td>
		</tr>
	</tbody>
</table>

studying left ventricular reverse remodeling by aortic debanding in rodents

为了更好地了解左心室 （LV） 反向改造 （RR），我们描述了一个啮齿动物模型，其中，在主动脉带引起的 LV 改造后，小鼠在切除主动脉收缩后接受 RR。在本文中，我们描述了一个分步手术，在小鼠中执行微创手术主动脉去除。回声心动图随后用于评估LV改造和RR期间心脏肥大和功能障碍的程度，并确定主动脉脱体的最佳时机。在协议结束时，对心脏功能进行了末期血液动力学评估，并收集了样本进行组学研究。我们表明，去除与手术存活率70-80%有关。此外，在解散两周后，心室后负荷的显著减少会触发心室肥大的回归（+20%）和纤维化（+26%），通过左心室填充和末期舒张压力（E/e'和LVEDP）的正常化评估舒张功能障碍的恢复。主动脉去除是研究啮齿动物LV RR的有用实验模型。因此，心肌恢复的范围在受试者之间是可变的，这模仿了临床环境中发生的RR的多样性，如主动脉瓣置换。我们的结论是，主动脉带状/脱体模型是解开对RR机制的新见解的宝贵工具，即心脏肥大的回归和舒张功能障碍的恢复。

术后和后期生存 带状手术的术外存活率为80%，第一个月的死亡率一般为20%<。如前所述，去除手术的成功在很大程度上取决于以前的手术的侵入性。经过学习曲线后，去除程序期间的死亡率约为 25%。对于这个死亡帐户大多是在手术过程中死亡，包括主动脉或左中庭破裂（在老鼠，生存率在两个外科手术更高）。
主动脉带和心肌重塑 主动脉收缩的...

Watch this Scientific Journal Video about Studying Left Ventricular Reverse Remodeling by Aortic Debanding in Rodents at JoVE.com

Studying Left Ventricular Reverse Remodeling by Aortic Debanding in Rodents

在这里，我们描述了在成熟的主动脉收缩模型中手术主动脉脱体的分步协议。这个过程不仅允许研究左心室反向改造和肥大回归背后的机制，而且还可以测试可能加速心肌恢复的新疗法方案。

研究左心室反向改造由主动脉脱体在啮齿动物

To better understand the left ventricular (LV) reverse remodeling (RR), we describe a rodent model wherein, after aortic banding-induced LV remodeling, ...

主动脉去除是研究啮齿动物心肌反向改造的有用实验模型，也是对心肌细胞回归和等离子体功能恢复机制的令人陶醉的新见解。这些金属有助于 VIVO 中批判反向建模的演变，以及在疾病进展期间不同时间点收集生物样本，用于体外和分子研究。该技术模拟心肌改造和反向改造的进展，并允许探索心肌恢复不完整的机制。该协议提供了对长期压力超载相关病理（如眼部自命不忠或主动脉狭窄）中完全和不完整的心肌反向改造的机制和线的见解。将骨质疏松管连接到呼吸机，启动机械通风，并将呼吸机调整为每分钟 160 次呼吸的频率和每公斤 10 毫升的潮汐体积。在开始带状手术之前，确认六到八周大的C57黑六只小鼠对疼痛反射缺乏反应，刮胡子，将脱毛霜从颈线涂到小鼠的中胸水平，并将软膏涂在动物的眼睛上。放置直肠探头和氧化计，用于监测温度血液氧化和心率。将鼠标放在加热垫上的右侧十进制，并将四肢粘在磁固定器缩回系统上，以便在手术过程中保持动物处于正确的位置。然后，用连续70%的酒精和波维酮-碘溶液擦洗胸部，然后打开皮肤肌肉和肋骨，以执行带到主动脉。在手术结束时，将波维酮碘溶液应用于皮肤缝合部位，每天两次进行镇痛，持续两到三天，然后注射无菌盐水内皮，以防止手术期间出现严重出血时的脱水，并将动物置于孵化器中，进行监测，直到完全康复。带状手术后七周，在一半的带状动物中轻轻解剖主动脉周围的组织、粘附和纤维化，直到收缩变得可见。仔细解剖主动脉，将缝合物与容器分离，然后用剪刀切割缝合线。请注意，在结合或恶魔期间，斧头蓝蓝水的长期渗透可能导致肺水肿，炎症通路的过度激活。使用尽可能少的缝合次数，用简单中断或连续的六条旧聚丙烯缝合物关闭胸壁，并以连续缝合模式用六O丝聚丙烯缝合皮肤，然后按照演示进行术后护理。手术后每两到三周将毛皮从颈线移到胸部中部，并按证明将动物放在超平位置的加热垫上，将心电图电极放在暴露的皮肤上，确认存在良好的心电图痕迹和每分钟 300 至 350 次心率。监测温度，并将回声凝胶涂抹在胸部区域。将鼠标放在后部重新任职，稍微向右转，并使用适当的设置启动回声心动图。接下来，将超声波探针定位在胸腔上，并评估整个主动脉的压力梯度。记录主动脉的二维引导图像，显示上升主动脉收缩的存在或缺失，以解剖学上可视化带状和去除的功效。要评估肥大，将探针放置在左心室短轴的状肌肉水平，并按下模式跟踪按钮，可视化左心室前壁、左心室直径和左心室后壁的二甲酰和收缩。然后评估收缩功能，并根据标准协议计算弹射分数和分数缩短。要评估舒张功能，使用在线粒体传单正上方的非典型四室视图确定早期和叶状线粒体流速的后波多普勒的峰值，并使用后组织多普勒成像和 apical 四室视图记录横向线环心肌早期舒张和峰值收缩速度。主动脉收缩的成功可以通过左心室和收缩压力的增加来验证。而由多普勒主动脉流速大于2.5米每秒。与沙姆操作的动物相比，弯曲诱导的左心室肥大表现为左心室质量增加和舒张功能受损，这表现在更高的填充压力和左心室末端舒张压。从结构上讲，七周的主动脉带诱发严重的心肌细胞肥大和纤维化。在我接受解散时，主动脉狭窄的成功去除可以通过回声多普勒速度来验证。整体去除会促进 LV 质量评估的后载和左心室肥大显著减少。此外，还观察到舒张功能正常化。当你仔细玩它避免长期的道德串通，以保证血液氧合保持在90%以上这个最低模型允许我们研究我的心脏改造和反向改造在疾病进展的不同时间点，甚至唤醒每个时间点最不良的调节路径。此程序允许评估有前途的药物的治疗潜力，并研究几个共病在心肌反向重塑和恢复的影响。

Research

JoVE Journal

Medicine

Reverse Total Shoulder Arthroplasty

反向共肩关节置换术

Reverse total shoulder arthroplasty was initially approved for use in rotator cuff arthropathy and well as chronic pseudoparalysis without arthritis in ...

科研

医学

Reduction in Left Ventricular Wall Stress and Improvement in Function in Failing Hearts using Algisyl-LVR

左室壁应力和改进功能衰竭心脏使用Algisyl LVR减少

Injection of  Algisyl-LVR, a treatment under clinical development, is intended to treat  patients with dilated cardiomyopathy. This treatment was recently ...

Retinal Detachment Model in Rodents by Subretinal Injection of Sodium Hyaluronate

视网膜脱离模型鼠害由透明质酸钠的视网膜下注射

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

研究胰腺癌干细胞特性开发新的治疗策略

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Transthoracic Speckle Tracking Echocardiography for the Quantitative Assessment of Left Ventricular Myocardial Deformation

经胸斑点跟踪超声心动图左室心肌变形的定量评价

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

重复性的动脉剥脱损伤腹主动脉阻断在小鼠模型

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

Full-root Aortic Valve Replacement by Stentless Aortic Xenografts in Patients with Small Aortic Roots

全根主动脉瓣置换主动脉瓣狭窄主动脉异种移植患者小主动脉根

In patients with small aortic roots who need an aortic valve replacement with biological valve substitutes, the implantation of the stented pericardial ...

Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy

小鼠横主动脉狭窄诱导左心室肥厚的技术研究

Transverse aortic constriction (TAC) in mice is one of the most commonly used surgical techniques for experimental investigation of pressure ...

Utilizing Percutaneous Ventricular Assist Devices in Acute Myocardial Infarction Complicated by Cardiogenic Shock

经皮心室辅助装置治疗急性心肌梗死并发心源性休克

Cardiogenic shock is defined as persistent hypotension, accompanied by evidence of end organ hypo-perfusion. Percutaneous ventricular assist devices ...

Use of a Percutaneous Ventricular Assist Device/Left Atrium to Femoral Artery Bypass System for Cardiogenic Shock

使用经皮心室辅助装置/左心房至股动脉旁路系统治疗心源性休克

The left atrial to femoral artery bypass (LAFAB) system is a mechanical circulatory support (MCS) device used in cardiogenic shock (CS) that bypasses the ...