Name: real-time pressure-volume analysis of acute myocardial infarction in mice
Uploaded: 2018-07-02T13:30:00
Description: Watch this Scientific Journal Video about Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice at JoVE.com

作者承认以下资金来源: 否则 Kröner-费森尤斯-基金会 (Tienush Rassaf);Hans 和格蒂 Fischer 基金会 (Tienush Rassaf), 补助金从医学教职员, 大学杜伊斯堡-埃森, 德国 (Tienush Rassaf, 米歇尔);Grimmke-基金会 (基督 Rammos)。

所有的实验都是按照和遵守所有相关的条例 (《欧洲保护用于实验和其他科学目的的脊椎动物的公约》 (指令 2010/63/欧盟) 完成的, 动物护理在符合机构准则。所有实验都是在6月龄的雄性 C57BL/6JRj 小鼠身上进行的。
1. 准备
<ol><li>准备一个外科显微镜和一个加热垫, 以及一个直肠探针来监测体温。清洁和消毒所有手术器械。</li>
	<li>为血管结扎、5厘米6-0 聚丙烯螺纹和2毫米硅管?...

<ol>
	<li>Sanchis-Gomar, F., Perez-Quilis, C., Leischik, R., Lucia, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+coronary+heart+disease+and+acute+coronary+syndrome.">Epidemiology of coronary heart disease and acute coronary syndrome.</a> Annals of Translational Medicine. 4 (13), 256 (2016).</li><li>Anderson, J. L., Morrow, D. A. <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.">Acute Myocardial Infarction.</a> New England Journal of Medicine. 376 (21), 2053-2064 (2017).</li><li>McNamara, R. 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=Predicting+in-hospital+mortality+in+patients+with+acute+myocardial+infarction.">Predicting in-hospital mortality in patients with acute myocardial infarction.</a> Journal of the American College of Cardiology. 68 (6), 626-635 (2016).</li><li>Kurian, G. A., Rajagopal, R., Vedantham, S., Rajesh, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Role+of+Oxidative+Stress+in+Myocardial+Ischemia+and+Reperfusion+Injury+and+Remodeling:+Revisited.">The Role of Oxidative Stress in Myocardial Ischemia and Reperfusion Injury and Remodeling: Revisited.</a> Oxidative Medicine and Cellular Longevity. , (2016).</li><li>Turer, A. T., Hill, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+myocardial+ischemia-reperfusion+injury+and+rationale+for+therapy.">Pathogenesis of myocardial ischemia-reperfusion injury and rationale for therapy.</a> American Journal of Cardiology. 106 (3), 360-368 (2010).</li><li>Totzeck, M., Hendgen-Cotta, U. B., French, B. A., Rassaf, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+practical+approach+to+remote+ischemic+preconditioning+and+ischemic+preconditioning+against+myocardial+ischemia/reperfusion+injury.">A practical approach to remote ischemic preconditioning and ischemic preconditioning against myocardial ischemia/reperfusion injury.</a> Journal of Biological Methods. 3 (4), (2016).</li><li>Respress, J. L., Wehrens, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transthoracic+echocardiography+in+mice.">Transthoracic echocardiography in mice.</a> Journal of Visualized Experiments. (39), (2010).</li><li>Pacher, P., Nagayama, T., Mukhopadhyay, P., Batkai, S., Kass, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+cardiac+function+using+pressure-volume+conductance+catheter+technique+in+mice+and+rats.">Measurement of cardiac function using pressure-volume conductance catheter technique in mice and rats.</a> Nature Protocols. 3 (9), 1422-1434 (2008).</li><li>Zhang, B., Davis, J. P., Ziolo, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+catheterization+in+mice+to+measure+the+pressure+volume+relationship:+Investigating+the+Bowditch+effect.">Cardiac catheterization in mice to measure the pressure volume relationship: Investigating the Bowditch effect.</a> Journal of Visualized Experiments. (100), e52618 (2015).</li><li>Rossello, X., Hall, A. R., Bell, R. M., Yellon, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+Langendorff+perfused+isolated+mouse+heart+model+of+global+ischemia-reperfusion+injury:+Impact+of+ischemia+and+reperfusion+length+on+infarct+size+and+LDH+release.">Characterization of the Langendorff perfused isolated mouse heart model of global ischemia-reperfusion injury: Impact of ischemia and reperfusion length on infarct size and LDH release.</a> Journal of Cardiovascular Pharmacology and Therapeutics. 21 (3), 286-295 (2016).</li><li>Task Force on the management of ST-segment elevation acute myocardial infarction of the European Society of Cardiology (ESC). <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ESC+Guidelines+for+the+management+of+acute+myocardial+infarction+in+patients+presenting+with+ST-segment+elevation.">ESC Guidelines for the management of acute myocardial infarction in patients presenting with ST-segment elevation.</a> European Heart Journal. 33 (20), 2569-2619 (2012).</li><li>Shigemitsu, O., 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=Acute+myocardial+infarction+due+to+left+main+coronary+artery+occlusion.+Therapeutic+strategy.">Acute myocardial infarction due to left main coronary artery occlusion. Therapeutic strategy.</a> Japanese Journal of Thoracic and Cardiovascular Surgery. 50 (4), 146-151 (2002).</li></ol>

对急性心肌梗死的 lv 血流动力学进行光电监测, 是实时评价心源性休克和左心室损伤的 lv 功能的一种新方法。PV 导管可以提供一个广泛的参数, 关于 LV 收缩和舒张功能。除了通常由超声心动图或 MRI (腔容积, EF, 中风体积和心脏输出) 获得的 lv 容积参数外, PV 分析通过同时提供 lv 收缩的措施, 产生一个更完整的 lv 功能剖面性能 (收缩力 dp/dt, 中风工作) 和 LV 遵约 (dp/dt, 头) 作为一个参数的舒张...

心血管疾病是西方文明中最常见的死亡原因1。急性心肌梗死是一个关键事件, 这是与高急性和慢性死亡率2。即使通过急诊经皮冠状动脉介入术 (PCI) 实现血管重建, 死亡率仍然很高, 特别是在48小时后, 急性心肌梗塞患者出现症状后3。急性左心室功能降低引起的心源性休克是3例住院死亡率的主要原因。这种早期的 LV 功能降低是由于缺血再灌注后心肌损伤所致。这种所谓的缺血/再灌注 (I/R) 损伤是介导的细胞代谢物的变化, 如夸大产生的活性氧物种4,5。
为了探索可能的保护机制, 导致在临床前设置的心肌损害减少, 可靠的鼠标模型是必不可少的, 包括评估的方法后, 左/R LV 功能6。在这一背景下, 经胸超声心动图7和磁共振成像 (MRI)6被广泛用于功能分型8,9。然而, 这些方法不适用于评估严重的 lv 功能障碍和心源性休克的持续急性心肌梗死, 不能直接显示有关 LV 压力的数据。在体外试验中使用隔离心脏的 Langendorff 装置提供了有关早期 I./R 损伤10的潜在 pathomechanisms 的信息。这种方法是有限的, 因为它无法重现体内的适应性机制, 如调节自主神经系统或激素调节和酸碱平衡。目前尚无方法可用于完全功能分型的心源性休克和左心室功能障碍在持续性心肌 I./R 损伤。
一种同步的方法, 结合压力容积 (PV) 插管和瞬态手术左冠状动脉主干 (LCA) 闭塞可能是有益的, 但技术上的挑战。稳定的源于血流动力学在我/R 损伤是必不可少的有效的结果, 因为不稳定的麻醉或失血可能严重影响的结果。采用低压光伏插管和瞬态 LCA 阻断技术对分型损伤的血流动力学进行研究, 可为急性心肌梗死患者心源性休克和 LV 功能障碍提供新的见解, 并作为今后分析心肌。

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			<td height="26" style="height:27px;width:237px;">Hamilton Syringe 100 &#181;l needle</td>
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			<td>100&#181;l syringe with needle</td>
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real-time pressure-volume analysis of acute myocardial infarction in mice

急性心肌梗死可导致急性心力衰竭和心源性休克。血流动力学的评估对于评估任何针对急性左心室 (LV) 功能障碍的潜在治疗方法都是至关重要的。目前的成像方式 (例如, 超声心动图和磁共振成像) 有几个局限性, 因为 LV 压力的数据不能直接测量。lv 导管在冠状动脉闭塞小鼠中的应用, 可作为实时评价 lv 功能的一种新方法。
在手术开始时, 小鼠被麻醉后气管插管。对于 LV 插管, 右颈动脉通过中颈部切口暴露。导管被引入并放置在 LV 腔内。左开胸术进行, 左主干冠状动脉 (LCA) 结扎。为诱导再灌注, 缝线在45分钟后释放, 在任何时候都记录了压力容积数据。
结扎的 LCA 导致 LV 收缩功能下降, 这证明了30% 减少中风体积, LV 弹射分数 (EF) 和心脏输出。最大 dp/dt 作为一个参数的 LV 收缩力也显著降低, 舒张功能严重受损 (最小 dP/dt-40%)。再灌注在20分钟期间没有导致 LV 作用的完全恢复。
实时压力容积分析作为监测急性心肌梗死小鼠心功能的有效方法。保持稳定的麻醉和规范化的手术方法对确保有效的结果至关重要。急性心肌梗死的早期阶段对发病率和死亡率有重要意义, 该方法可为心肌新策略的临床前评价提供有益的借鉴。

经低压插管后, 45 分钟可逆性 LCA 结扎术后再灌注10分钟。PV 数据在任何时候都被记录下来 (图 1)。
通过获得特征 LV pv 图 (图 2A), 确定了 pv 导管的正确位置。LV 导管放置显示典型的心室压力范围, 至少 0-20 mmHg, 而在主动脉中的 PV 导管假放置将显示一个典型的动脉压力曲线, 最小压力为...

Watch this Scientific Journal Video about Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice at JoVE.com

Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice

小鼠急性心肌梗死诱发左心室 (LV) 功能的急性但不完全特征性改变。lv 导管在冠状动脉闭塞小鼠中的应用是一种实时评价 lv 功能的新方法。

小鼠急性心肌梗死的实时压力容积分析

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any ...

This method can help answer key questions in the field of cardiovascular research such as identifying early protective mechanisms in acute myocardia infraction. The main advantage of this technique is that an advanced hemodynamic characterization can be performed real time in mice undergoing myocardial infarction followed up by reperfusion. Endotracheal intubation and ischemia reperfusion surgery will be performed by Pia Stock, a biologist from our laboratory.After anesthetizing the animal, remove the neck and chest hair using electric clippers. Next, clean the skin with three alternating scrubs of Betadine and 70%alcohol. Now, using small surgical scissors, make a 10-millimeter longitudinal median incision five millimeters beneath the bottom lip and towards the sternum.Next, bluntly dissect the left and right part of the submandibular gland using forceps. Then, separate the muscle and fat tissue in the right paratracheal region to access the right commune corroded artery. Using curved forceps, carefully bluntly dissect the vessel to mobilize and separate five to 10 millimeters of it.Always avoid moving the vagus nerve in the corroded body, as this can lead to severe hypotension and bradycardia. After the vessel mobilization, pass the two prepared 5/0 silk sutures under the vessel. Then ligate the distal vessel using a tight knot, and place a loose knot on the proximal suture where the catheter will go.Next, place a hemostat vascular clamp on the proximal end of the exposed vessel, proximal to the suture. This will be used to reversibly block blood flow. Now, using microscissors, open the vessel with the wedge-shaped incision one millimeter proximal to the cranial knot.A small drop of blood should be seen. Then, stretch the incision and insert the catheter 10 millimeters into the vessel. Now, start recording the data collected by the catheter probe.Next, extract the vascular clamp, then add one or two drops of saline into the incision to help slide the catheter another 10 millimeters down the vessel. Then, tighten the knot to prevent blood reflux alongside the catheter without locking the catheter in place. Now, gently continue advancing the catheter until the pressure analysis shows an arterial blood pressure profile when the catheter has reached the aorta.At the aortic valve, light resistance and pull synchronized movement should also be felt. When trying to advance the catheter through the aortic valve, it can help to pull it back five millimeters and advance again. Proper left ventricular catheterization will be indicated by a change in diastolic pressure.Any change in volume is also an indicator of advancing the sensor to the left ventricle. Once certain that the catheter is in the left ventricle, fasten the proximal knot more tightly to prevent it from moving. After the catheterization, prepare the 45 minute ischemia.First, incise the skin from the caudal sternum to the left axilla. Then proceed with the blunt preparation of the two muscle layers to expose the ribs. Next, open the thorax with an incision between the third and fourth left rib.Make use of surgical hooks to access the pericardium, and then resect the pericardium covering the heart. Before continuing with the left main coronary artery ligation, wait 30 seconds without touching the animal to record pressure and volume data for a valid analysis. Now, identify the left main coronary artery.It emerges under the left oracle and descends at the left side of the heart towards the apex. Then, loop a 6/0 polypropylene suture around the artery two millimeters under the left oracle. Next, place a small silicon tube under the loop and secure it with a knot.If the distal myocardium starts to turn gray, then the left main coronary artery has been occluded. To proceed, cut the suture at one millimeter length, and then release the surgical hooks and manually close the muscle layers above the incision. Then wait 45 minutes while continuously recording pressure and volume data.After 45 minutes, reopen the incision and remove the silicon tube to induce reperfusion. Then record data for another 20 minutes. When the heart turns back to being red, the reperfusion is successful.After the reperfusion period, loosen the knot and extract the catheter while quickly fastening the knot again to prevent loss of blood if blood is needed for further studies. After completing the calibration steps, which are detailed in the text protocol, perform a software-guided data analysis. For the baseline data, highlight at least 10 cycles taken before the passing of the catheter into the aortic valve.As needed, exclude any cycles with deviations due to ventilation or manipulation. Next, analyze 10 cycle blocks on five minute intervals through the ischemia and reperfusion. The various parameters calculated by this analysis are sufficient for characterization of the left ventricular function.At the end of the experiment, after the catheter has been retracted from the left ventricle, take a final analysis of the pressure data. Following the protocol, after the left ventricular catheterization, reversible left main coronary artery ligation was performed for 45 minutes followed by 20 minutes of reperfusion. Correct placement of the volume and pressure catheter was confirmed by obtaining the characteristic left ventricular pressure-volume graph.Successful occlusion of the left main coronary artery was visually confirmed by blanching of the distal left ventricular myocardium. After the left main coronary artery occlusion, pressure and volume data was analyzed on five minute intervals. Pressure data showed no changes in maximum left ventricular systolic pressure, indicating preserved peripheral perfusion and stable anesthesia.Analysis of left ventricular volume revealed that during the early phase of the ischemia, there was a significant decrease in ejection fraction and in absolute stroke volume. The maximum change in pressure, a parameter of left ventricular contractility, showed a 30%reduction in mice undergoing myocardial ischemia. The minimum change in pressure significantly decreased as well, indicating impaired left ventricular compliance.After watching this video, you should have a good understanding on how to characterize left ventricular systolic and diastolic function in mice undergoing acute myocardial infarction using pressure-volumes catheter technique.

Research

JoVE Journal

Medicine

Acute Myocardial Infarction in Rats

大鼠急性心肌梗死

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular ...

科研

医学

Induction of Myocardial Infarction in Adult Zebrafish Using Cryoinjury

在成年斑马鱼心肌梗死的感应使用Cryoinjury

The mammalian heart is incapable of significant regeneration following an acute injury such as myocardial infarction1. By contrast, urodele amphibians and ...

MRI and PET in Mouse Models of Myocardial Infarction

MRI和PET心肌梗死小鼠模型

Myocardial infarction is one of the leading causes of death in the Western world. The similarity of the mouse heart to the human heart has made it an ...

Myocardial Infarction and Functional Outcome Assessment in Pigs

心肌梗死和功能预后评估的猪

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One ...

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 ...

Primary Outcome Assessment in a Pig Model of Acute Myocardial Infarction

急性心肌梗死的猪模型主要成果评估

Mortality after acute myocardial infarction remains substantial and is associated with significant morbidity, like heart failure. Novel therapeutics are ...

Myocardial Infarction in Neonatal Mice, A Model of Cardiac Regeneration

心肌梗死新生小鼠，心肌再生的模型

Myocardial infarction induced by coronary artery ligation has been used in many animal models as a tool to study the mechanisms of cardiac repair and ...

Digital PCR for Quantifying Circulating MicroRNAs in Acute Myocardial Infarction and Cardiovascular Disease

数字 PCR 在急性心肌梗死和心血管疾病中量化循环 MicroRNAs 的临床观察

Circulating serum microRNAs (miRNAs) have shown promise as biomarkers for the cardiovascular disease and acute myocardial infarction (AMI), being released ...

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 ...

A Modified Simple Method for Induction of Myocardial Infarction in Mice

一种改良的小鼠心肌梗死诱导简易方法

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI ...