Name: left coronary artery ligation: a surgical murine model of myocardial infarction
Uploaded: 2022-08-09T14:45:00
Description: Watch this Scientific Journal Video about Left Coronary Artery Ligation: A Surgical Murine Model of Myocardial Infarction at JoVE.com

这项工作得到了美国国立卫生研究院拨款（R01HL143967，R01HL142629，R01AG069399和R01DK129339），AHA转型项目奖（19TPA34910142），AHA创新项目奖（19IPLOI34760566）和ALA创新项目奖（IA-629694）（PD）的支持。

本研究方案由匹兹堡大学机构动物护理和使用委员会（IACUC）审查和批准。8只（假n = 4和MI n = 4）体重24-30g的1岁雌性C57BL / 6J小鼠用于这些实验。大约100%和至少80%的小鼠分别在前24小时和28天内存活。
1.小鼠的准备和气管插管
<ol><li>将珠子灭菌器（见 材料表）预热至250°C，并将高压灭菌的手术器械放入其中几分钟。</li>
	<li>在含有3%异氟醚和1L / min氧气...

<ol>
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(150), e59591 (2019).</li><li>Muthuramu, I., Lox, M., Jacobs, F., De Geest, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Permanent+ligation+of+the+left+anterior+descending+coronary+artery+in+mice:+a+model+of+post-myocardial+infarction+remodelling+and+heart+failure.">Permanent ligation of the left anterior descending coronary artery in mice: a model of post-myocardial infarction remodelling and heart failure.</a> Journal of Visualized Experiments. (94), e52206 (2014).</li><li>Pistner, A., Belmonte, S., Coulthard, T., Blaxall, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+echocardiography+and+ultrasound+imaging.">Murine echocardiography and ultrasound imaging.</a> Journal of Visualized Experiments. (42), e2100 (2010).</li><li>Li, L., 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=Assessment+of+cardiac+morphological+and+functional+changes+in+mouse+model+of+transverse+aortic+constriction+by+echocardiographic+imaging.">Assessment of cardiac morphological and functional changes in mouse model of transverse aortic constriction by echocardiographic imaging.</a> Journal of Visualized Experiments. (112), e54101 (2016).</li><li>Vasamsetti, S. B., 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=Sympathetic+neuronal+activation+triggers+myeloid+progenitor+proliferation+and+differentiation.">Sympathetic neuronal activation triggers myeloid progenitor proliferation and differentiation.</a> Immunity. 49 (1), 93-106 (2018).</li><li>Reichert, 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=Murine+left+anterior+descending+(LAD)+coronary+artery+ligation:+an+improved+and+simplified+model+for+myocardial+infarction.">Murine left anterior descending (LAD) coronary artery ligation: an improved and simplified model for myocardial infarction.</a> Journal of Visualized Experiments. (122), e55353 (2017).</li><li>Alwardt, C. M., Redford, D., Larson, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=General+anesthesia+in+cardiac+surgery:+a+review+of+drugs+and+practices.">General anesthesia in cardiac surgery: a review of drugs and practices.</a> The Journal of Extra-Corporeal Technology. 37 (2), 227-235 (2005).</li><li>Kolk, M. V., 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=LAD-ligation:+a+murine+model+of+myocardial+infarction.">LAD-ligation: a murine model of myocardial infarction.</a> Journal of Visualized Experiments. (32), e1438 (2009).</li><li>Wu, Y., Yin, X., Wijaya, C., Huang, M. -. H., McConnell, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+myocardial+infarction+in+rats.">Acute myocardial infarction in rats.</a> Journal of Visualized Experiments. (48), e2464 (2011).</li><li>Kalogeris, T., Baines, C., Krenz, M., Korthuis, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+six-cell+biology+of+ischemia/reperfusion+injury.">Chapter six-cell biology of ischemia/reperfusion injury.</a> International Review of Cell and Molecular Biology. , 229-317 (2012).</li><li>Scofield, S. 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(117), e54814 (2016).</li><li>Yan, X., 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=Temporal+dynamics+of+cardiac+immune+cell+accumulation+following+acute+myocardial+infarction.">Temporal dynamics of cardiac immune cell accumulation following acute myocardial infarction.</a> Journal of Molecular and Cellular Cardiology. 62, 24-35 (2013).</li><li>Xu, Q., 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=Protective+effects+of+fentanyl+preconditioning+on+cardiomyocyte+apoptosis+induced+by+ischemia-reperfusion+in+rats.">Protective effects of fentanyl preconditioning on cardiomyocyte apoptosis induced by ischemia-reperfusion in rats.</a> Brazilian Journal of Medical and Biological Research. 50 (2), 5286 (2017).</li><li>Leuschner, F., 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=Rapid+monocyte+kinetics+in+acute+myocardial+infarction+are+sustained+by+extramedullary+monocytopoiesis.">Rapid monocyte kinetics in acute myocardial infarction are sustained by extramedullary monocytopoiesis.</a> Journal of Experimental Medicine. 209 (1), 123-137 (2012).</li><li>Leuschner, F., 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=Silencing+of+CCR2+in+myocarditis.">Silencing of CCR2 in myocarditis.</a> European Heart Journal. 36 (23), 1478-1488 (2015).</li><li>Dutta, 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=E-selectin+inhibition+mitigates+splenic+HSC+activation+and+myelopoiesis+in+hypercholesterolemic+mice+with+myocardial+infarction.">E-selectin inhibition mitigates splenic HSC activation and myelopoiesis in hypercholesterolemic mice with myocardial infarction.</a> Arteriosclerosis, Thrombosis, and Vascular Biology. 36 (9), 1802-1808 (2016).</li><li>Jiang, C., 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+modified+simple+method+for+induction+of+myocardial+infarction+in+mice.">A modified simple method for induction of myocardial infarction in mice.</a> Journal of Visualized Experiments. 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MI的小鼠模型在心血管研究实验室中越来越受欢迎，本研究描述了一种可重复且临床相关的MI模型。该协议以多种方式改进了LCA连接过程。首先，避免使用可注射的术前麻醉剂，例如甲苯噻嗪/氯胺酮或戊巴比妥钠14，15 。仅使用异氟醚麻醉，这有助于提高动物存活率（术后28天存活率为>80%），最大限度地减少药物引起的并发症，并且与其他药物相比，心?...

心血管疾病是一个重大的公共卫生问题，每年夺走1790万人的生命，占全球死亡率的31%1。最普遍的心血管异常类型是冠心病，心肌梗塞（MI）是冠心病的主要表现之一2。心肌梗死通常是由脆弱斑块破裂引起的冠状动脉血栓闭塞引起的3.由此产生的缺血导致受影响的心肌发生深刻的离子和代谢变化，以及收缩功能的快速下降。心肌梗死导致心肌细胞死亡，可进一步导致心室功能障碍和心力衰竭4。
由于从MI5患者获得的组织稀缺，对患者心肌梗死的研究有限。因此，MI的小鼠模型可用于研究疾病机制以及开发潜在的治疗靶点。目前可用的MI小鼠模型包括不可逆缺血模型（LCA和消融方法）和再灌注模型（缺血/再灌注，I / R）6。小鼠左冠状动脉（LCA）的永久性结扎是最常用的方法，它模仿患者7，8，9中MI的病理生理学和免疫学。永久性心肌梗死也可以通过消融方法诱发，包括电损伤或冷冻损伤。消融方法能够在精确位置10产生均匀大小的梗死。另一方面，瘢痕形成、梗死形态和分子信号传导机制可能因消融方法而异10，11。鼠I/R方法是另一个重要的MI模型，因为它代表了再灌注治疗的临床场景12。I/R 模型存在各种挑战，例如梗死大小可变、难以区分初始损伤的反应和再灌注6。
虽然被广泛使用，但LCA结扎方法与低生存率和术后疼痛有关13。该协议展示了LCA结扎的小鼠手术MI模型，该模型涉及小鼠的准备和插管，LCA结扎，术后护理和MI的验证。 该方法不使用侵入性气管切开术14，而是采用气管插管。通过使用喉镜照亮口咽来对动物进行插管，使手术更容易、更安全、创伤更小15.在整个过程中，小鼠保持在呼吸机支持下并在异氟醚麻醉下。此外，进行超声心动图和Masson三色染色分别评估心肌梗死后的心脏功能和心肌纤维化。总体而言，该方法提供了一种可靠且可重复的MI手术鼠模型，可用于研究MI后的病理生理学和炎症。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>22 G catheter needle</td>
			<td>Exel INT</td>
			<td>26741</td>
			<td>Thoracentesis</td>
		</tr>
		<tr>
			<td>24 G catheter needle</td>
			<td>Exel INT</td>
			<td>26746</td>
			<td>Endotracheal intubation</td>
		</tr>
		<tr>
			<td>4-0 nylon suture</td>
			<td>Covetrus</td>
			<td>29263</td>
			<td>Suturing of muscles and skin</td>
		</tr>
		<tr>
			<td>8-0 nylon suture</td>
			<td>S&#38;T</td>
			<td>3192</td>
			<td>Ligation of LAD</td>
		</tr>
		<tr>
			<td>Anesthetic Vaporizers</td>
			<td>Vet equip</td>
			<td>VE-6047</td>
			<td>Anesthetic support</td>
		</tr>
		<tr>
			<td>Animal physiology monitor</td>
			<td>Fujifilm</td>
			<td>VEVO 3100</td>
			<td>Monitor heart rate,respiration rate and body temperature</td>
		</tr>
		<tr>
			<td>Betadine solution</td>
			<td>PBS animal health</td>
			<td>11205</td>
			<td>Antispetic</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Covetrus</td>
			<td>55175</td>
			<td>Analgesic</td>
		</tr>
		<tr>
			<td>Disecting microscope</td>
			<td>OMANO</td>
			<td>OM2300S-V7</td>
			<td>Binocular</td>
		</tr>
		<tr>
			<td>Electric razor</td>
			<td>Wahl</td>
			<td>79300-1001M</td>
			<td>Shaving</td>
		</tr>
		<tr>
			<td>Electrode gel</td>
			<td>Parker Laboratories</td>
			<td>W60698L</td>
			<td>Electrically conductive gel</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Decon Laboratories</td>
			<td>22-032-601</td>
			<td>Disinfectant</td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>FST</td>
			<td>11065-07</td>
			<td>Stainless Steel</td>
		</tr>
		<tr>
			<td>Gauze</td>
			<td>Curity</td>
			<td>CAR-6339-PK</td>
			<td>Sterile</td>
		</tr>
		<tr>
			<td>Heat lamp</td>
			<td>Satco</td>
			<td>S4998</td>
			<td>Post surgery care</td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Kent scientific</td>
			<td>Surgi-M</td>
			<td>Temperature control</td>
		</tr>
		<tr>
			<td>Hot Bead sterilizer</td>
			<td>Germinator 500</td>
			<td>11503</td>
			<td>Sterilization of surgical instrument</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Covetrus</td>
			<td>29405</td>
			<td>Anesthesia</td>
		</tr>
		<tr>
			<td>Masson&#8217;s trichrome staining kit</td>
			<td>Thermoscientific</td>
			<td>87019</td>
			<td>Measurement of cardiac Fibrosis</td>
		</tr>
		<tr>
			<td>Micro Needle Holder</td>
			<td>FST</td>
			<td>12500-12</td>
			<td>Stainless Steel</td>
		</tr>
		<tr>
			<td>Micro scissors</td>
			<td>FST</td>
			<td>15000-02</td>
			<td>Stainless Steel</td>
		</tr>
		<tr>
			<td>Ophthalmic ointment</td>
			<td>Dechra</td>
			<td>Puralube Vet</td>
			<td>Sterile occular lubricant</td>
		</tr>
		<tr>
			<td>Scanning Gel</td>
			<td>Parker Laboratories</td>
			<td>Aquasonic 100</td>
			<td>Aqueous ultrasound transmission gel</td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>FST</td>
			<td>14060-11</td>
			<td>Stainless Steel</td>
		</tr>
		<tr>
			<td>Small Animal Laryngoscope</td>
			<td>Penn-Century</td>
			<td>Model LS-2-M</td>
			<td>Illuminating the oropharynx</td>
		</tr>
		<tr>
			<td>Small animal ventilator</td>
			<td>Harvard apparatus</td>
			<td>557058</td>
			<td>Ventilator support</td>
		</tr>
		<tr>
			<td>Surgical light</td>
			<td>Cole parmer</td>
			<td>41723</td>
			<td>Illuminator Width (in): 7</td>
		</tr>
		<tr>
			<td>Vevo 3100 preclinical imaging platform</td>
			<td>Fujifilm</td>
			<td>VEVO 3100</td>
			<td>Echocardiography</td>
		</tr>
		<tr>
			<td>VevoLAB software</td>
			<td>Fujifilm</td>
			<td>VevoLAB 3.2.6</td>
			<td>Echocardiography data analysis</td>
		</tr>
	</tbody>
</table>

left coronary artery ligation: a surgical murine model of myocardial infarction

缺血性心脏病和随后的心肌梗塞（MI）是美国和世界各地死亡的主要原因之一。为了探究心肌梗死后的病理生理变化并设计未来的治疗方法，需要心肌梗死的研究模型。小鼠左冠状动脉（LCA）的永久性结扎是研究心肌梗死后心脏功能和心室重塑的流行模型。在这里，我们描述了一种侵入性较小、可靠且可重复的手术鼠 MI 模型，通过永久连接 LCA。我们的手术模式包括易于逆转的全身麻醉、不需要气管切开术的气管插管和开胸术。应进行心电图和肌钙蛋白测量以确保心肌梗死。 心肌梗死后第 28 天的超声心动图可识别心功能和心力衰竭参数。心脏纤维化的程度可以通过Masson的三色染色和心脏MRI进行评估。该心肌梗死模型有助于研究心肌梗死后的病理生理和免疫学改变。

图1显示了假（图1A）和MI（图1B）小鼠超声心动图评估期间的代表性主动心电图和呼吸信号。在获取超声心动图数据之前，验证主动心电图和呼吸信号很重要。 图2显示了LCA结扎后28天心脏功能参数的超声心动图测量。图 2 显示了假心脏（图 2A）和 MI（图 2B</s...

Watch this Scientific Journal Video about Left Coronary Artery Ligation: A Surgical Murine Model of Myocardial Infarction at JoVE.com

Left Coronary Artery Ligation: A Surgical Murine Model of Myocardial Infarction

这里介绍的是小鼠左冠状动脉永久结扎的外科手术。该模型可用于研究心肌梗死后的病理生理学和相关炎症反应。

左冠状动脉结扎术：心肌梗死的外科小鼠模型

Ischemic heart disease and subsequent myocardial infarction (MI) is one of the leading causes of mortality in the United States and around the world. In ...

手术模型对小鼠左动脉的永久结扎可用于研究心肌梗死后的病理生理学和相关炎症反应。该方法提供了一种可靠且可生产的MI小鼠模型，该模型简单，安全且侵入性较小。首先，将珠子灭菌器预热至250摄氏度，然后将高压灭菌的手术器械放入其中几分钟。通过检查对坚硬的脚趾捏的反应来确保鼠标的麻醉深度。称量小鼠以估计术前镇痛药物丁丙诺啡的剂量，并腹腔注射药物。使用电动剃须刀修剪胸部左侧的皮毛。对手术部位进行消毒后，将鼠标仰卧放在倾斜的板上。使用分别连接到上门牙和胶带的松紧带固定鼠标的头部和四肢。在麻醉下将无菌眼科润滑剂涂抹在眼睛上，以防止干燥。打开下颌，轻轻地将舌头拉出口腔。通过使用喉镜照亮口腔咽来识别喉部的开口。从 24 号导管针上切下约 0.5 厘米，然后将钝针插入塑料防护罩中。将带有塑料护罩的钝针引导至气管中。取出针头，将塑料护罩留在气管中。将呼吸机设置为每分钟137次的呼吸频率，针对本研究中使用的小鼠和潮气量0.18闭合能力进行了优化。将呼吸器管连接到导管护罩，并通过寻找与呼吸机同步的胸部运动来确认插管正确。断开呼吸管与导管护罩的连接，并将动物仰卧在预热的温控手术板上。将鼠标重新连接到呼吸机。消毒手术部位后，使用一对镊子轻轻抬起皮肤，并使用一把手术剪刀沿着左胸大肌和小胸肌之间的线做一个小的皮肤横向切口。用镊子和解剖剪刀分开下面的胸大肌。在第三肋间隙切开后，使用牵开器轻轻地将肋骨分开，露出左心室。将心包脂肪移到一边，找到从左心房边缘向心脏顶点延伸的左冠状动脉。传球8-0在持针器的帮助下在左冠状动脉下进行尼龙缝合。左冠状动脉打一个双结，然后打第二个结。取下牵开器并将 22 号导管针插入胸腔。取下针头，将塑料护罩的尖端留在胸腔中。使用4-0尼龙缝合线关闭胸腔。将注射器连接到 22 号塑料防护罩，并通过轻轻按压胸部以建立负气压来缓慢去除困在胸腔中的多余空气。取下塑料防护罩。关闭异氟醚供应，并将鼠标放在供氧的呼吸机上。小鼠开始自主呼吸，关闭呼吸机。将鼠标放在加热灯下并监视，直到它处于唤醒状态。在动物恢复足够的意识以维持胸骨卧位之前，不应无人看管。每天监测鼠标是否有任何疼痛或不适的迹象。手术后每 6 至 8 小时继续腹腔注射丁丙诺啡，再注射两天。在获取超声心动图数据之前，对主动心电图和呼吸信号进行了验证。在左冠状动脉结扎术后28天注意到心脏功能参数的超声心动图测量。观察SHAM和MI心脏的腓骨短轴视图的M模式图像。心肌梗死图像显示左冠状动脉结扎术后心壁运动缺陷。与SHAM组相比，MI组观察到心力衰竭指标，例如左心室质量增加，射血分数降低和心输出量减少。Masson 下、中、上脑室部分的三色染色显示梗死心脏中的胶原蛋白染色增加，表明纤维化增强。在大多数情况下，动脉是不可行的。然而，在左心房边缘以下 2 至 4 毫米处结扎心肌组织可有效结扎动脉。

Research

JoVE Journal

Medicine

Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction

结扎冠状动脉内注射在小鼠模型梗死

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo.  With the advent of genetic modifications to ...

科研

医学

Modified Technique for Coronary Artery Ligation in Mice

改性技术在小鼠冠状动脉结扎

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

心肌缺血再灌注损伤的通过左前降支结扎小鼠模型

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

在诱导小鼠心肌梗死的微创手术过程

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

左冠状动脉前降支的小鼠结扎：对心肌梗死后重塑和心衰模型

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

左心房，主动脉和肺动脉的超声心动图小鼠

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic ...

Murine Left Anterior Descending (LAD) Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

Murine Left Anterior Descending LAD Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

鼠左前降支（LAD）冠状动脉结扎：改进和简化模型的心肌梗死

Ischemic heart disease (IHD), or acute coronary syndrome (ACS), is one of the leading causes of death in the United States. IHD is characterized by ...

Direct Re-implantation of Left Coronary Artery into the Aorta in Adults with Anomalous Origin of Left Coronary Artery from the Pulmonary Artery (ALCAPA)

Direct Re-implantation of Left Coronary Artery into the Aorta in Adults with Anomalous Origin of Left Coronary Artery from the Pulmonary Artery ALCAPA

左冠状动脉直接再注入与从肺动脉左冠状动脉起源异常成人主动脉（ALCAPA）

Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital anomaly which is one of leading causes of myocardial ...

Murine Myocardial Infarction Model using Permanent Ligation of Left Anterior Descending Coronary Artery

使用左前部降冠动脉的永久结扎的月心肌梗死模型

Myocardial infarction (MI) and acute coronary diseases are among the most prominent causes of death in population with western lifestyle. The murine ...

Intramyocardial Transplantation of MSC-Loading Injectable Hydrogels after Myocardial Infarction in a Murine Model

Murine 模型心肌梗塞后 MSC 加载注射水凝胶的心内移植

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of ...