This work was supported by Interdisciplinary Centre for Clinical Research IZKF Aachen (junior research group to E.A.L.) within the faculty of Medicine at RWTH Aachen University. We are grateful Dr. Rusu and Ashley Christina Vourakis for critical review of the manuscript and Mrs. Roya Soltan for the professional help with immunohistochemistry staining.

注：本文所提出的实验也相应进行德国低，欧洲动物护理准则。动物繁殖研究所在实验动物科学，Universiy医院亚琛，德国的动物设施的监督下教授R.·托尔巴博士和A.托伊布纳（动物福利主任）的。 1.动物护理<ol><li>保持老鼠在一个专门的监护病房，确保正确的获得食物和专业兽医的控制和治疗。如果动物被移动或从外部购买的，请接受手术前保证一周的住宿。 </li></ol> 2.插管<ol><li>麻醉8-10周龄雄性C57BL / 6野生型小鼠，25-27克用腹膜内注射100mg / kg的氯胺酮和10mg / kg的甲苯噻嗪。监测麻醉通过脚趾捏反射的水平。放置在眼睛兽医软膏，以防止在操作过程中干。 </li><l我&gt;开始手术，按照你的机构的动物护理准则之前管理的疼痛治疗丁丙诺啡与0.1毫克/千克体重皮下注射。 </li><li>保证无菌条件的维护，以避免在手术期间的感染通过使用无菌的材料和工具。 </li><li>将麻醉小鼠中在加热的手术台仰卧位置。使用小的剃刀除去从两个腹的颈部区域和胸部的左半头发和消毒之前用切口70％的酒精。 <ol><li>执行使用手术剪刀在颈部的中心0.5cm的小正中切口。在皮肤下，经过2脂肪体用无菌弯钳，并通过覆盖肌肉的透明可视化立体显微镜下的气管。 </li></ol></li><li>利用体视显微镜（图1A）口头介绍插管插管插入视图下气管。区分金属套管救援人员到场唉透明的组织。并检查，位置和定位的操作在任何时刻（ 图1B）中。 </li><li>套管连接到小动物呼吸机和通风根据设置来调整生产准则（100-150微升和100-150之间的呼吸速率每分钟之间的潮气量）。 </li></ol> 3.心肌梗死感应<ol><li>在xyphoid和左axila之间的线的中间执行一个皮肤切口小于0.5cm。使用镊子肌肉层从底层的肋骨分开。 </li><li>通过使用小剪刀，直到胸腔打开17执行的肋之间有一个小切口。从下面的2 个和第一个更简单的方法编号：慢性梗死，执行切口的第 5肋间空间（ 图1C）和/或用于局部缺血/再灌注模型中，在第 4 次肋间隙（ 图1D） 3 日 </suP&gt;肋间，分别地）。 </li><li>将拉钩插入切口打开胸腔，并以可视化的心脏。 </li><li>小心取出心包，以防止过度纤维化的进程。 </li><li>可视化的左冠状动脉前降支（LAD）作为深定位淡红色容器。如果法援署不能进行可视化，考虑一些参考点，以增加可重复性。 <ol><li>对于慢性梗死模型中，放置结扎在心脏（耳廓和顶点之间）的腹侧的中间，具有作为参考的静脉， 如图1C所示 。绑定使用0/7丝线获得透前部和后部梗死动脉的两个分支。灰度颜色指示结扎的位置，如果需要（ 图1C）可以重复。 </li><li>对缺血/再灌注模型中，放置结扎耳廓下，经LAD的主体（ 图1D）。该LIGATURE位于在硅管，以保护容器的完整性。灰色的颜色表明梗死面积，应该出现在整个心脏（ 图1D）。广场上的排骨时间缝线在缺血期和滋润使用压缩，以避免组织干燥。缺血后，除去硅管，并切断与小剪刀缝线以可视化的再灌注。 </li></ol></li><li>旁在程序开始时所使用的麻醉剂和止痛剂（步骤2.1和2.2），用外科手术过程中的0.5％的异氟烷​​，以确保动物的适当的舒适度，或者跟随的机构的动物护理准则。 </li></ol> 4.缝合和恢复<ol><li>通过填充用温等渗盐水消除胸部残留空气。 </li><li>关闭胸部用3缝线0/6（ 见图2A和2B）。定位内侧缝线以90°的角度，以确保如ealed肋的闭合，如图2（ 图2A，B）。 </li><li>关闭肌肉层用2缝线（ 图2C）中，用3-4缝合线0/6（ 图2D）的皮肤。分别执行这些缝合线以得到合适的窗口进行进一步的超声心动图测量。 </li><li>断开呼吸机插管插管，并允许自主呼吸。对于以后的识别，标记使用本地系统（请动物福利工作人员从您所在的机构）的鼠标。 </li><li>在红灯放下鼠标左侧，直到它唤醒。不要让动物无人值守，直到它已经恢复了神志充足。不允许已经历手术，可以在其他动物的公司，直到完全恢复的动物。 </li><li>管理疼痛治疗丁丙诺啡与0.1毫克/千克体重，皮下注射，未来3天，按照你的机构的动物护理准则。 </li&gt; </ol>心肌梗死5.分析<ol><li>定期监测通过的超声心动图（ 图3A）装置中的心脏功能：射血分数，缩短分数，心输出量和心脏的尺寸。 </li><li>麻醉用腹膜内注射100mg / kg的氯胺酮和10mg / kg的甲苯噻嗪的动物。由于缺乏反射的确认正确麻醉手术前。 </li><li>打开胸腔，并切除心脏，将其放置在无菌PBS液洗涤广泛的剩余的血液。 </li><li>如果需要，直接从心脏通过避免梗死区域的损伤采集血液，或摘取心脏后，从胸腔。 </li><li>清洗后，停在diastola饱和氯化钾溶液中的心脏（与不孕过滤3M氯化钾PBS）。组织学分析固定在心脏10％福尔马林并继续步骤5.7。 </li><li>如果必要的话，测量汽车的可行性DIAC细胞埃文斯蓝/三苯基氯化四氮唑（TTC）染色。重建结扎在初始位置后，用灌注主动脉插管用200微升1％埃文斯蓝溶液的心脏，并在-20℃冷冻的心脏在一个小塑料袋，无需清洗。 <ol><li> 2小时后，用锋利的解剖刀执行5横向滑动，并培育它们在TTC溶液10-15分钟，在37℃下，所描述的制造。固定的幻灯片在10％福尔马林10分钟，并把它们的显微镜载玻片进行进一步的分析之间。 </li><li>嵌入心脏组织中的石蜡，通过尖端定位的心脏，进行横向切片。执行的5微米连续切片。收集前20段和丢弃在下一个300微米。继续部分协议直到二尖瓣水平已经达到（ 图3A，B）。连续切片，400微米间隔在整个心脏被收集并可以被染色˚F或定性和定量分析。 </li></ol></li><li>使用莫里的一步法染色6-8测量梗死面积。 </li><li>使用通常的免疫组织染色中连续切片分析血管形成，胶原蛋白含量或炎症细胞募集。 </li></ol>

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The authors have nothing to disclose.

在实验过程中，也有要注意一些关键点：插管，所述打开胸腔，并在LAD结扎。第一个关键步骤是experiements前动物的插管。许多团体正在使用用于固定鼠标和光源直接插入套管插入气管竖直支撑。该方法具有涉及插管进入气管插入正确的不确定性，是最容易出现故障的新手。制造一个小切口，套管的位置可在整个动作过程中被控制，从而降低了默认速率。此外，气管切开被超过，从而减少并发症，减少操作的时间。 下一关键步骤是胸腔的开口。胸骨正中代表了高风险的机动延缓动物的恢复。横向切开左暗示的2-3肋15的切割</SUP&gt;，导致缺陷的恢复和死亡率上升。我们使用的肋骨提供了最小的负担之间的小模型，离散切口。这些动物在手术后很快恢复，不存在缺陷或干扰愈合。下部间肋软骨空间被取为参考点。考虑到这一点，适当的和差异化的进入慢性缺血/再灌注模型结扎的地方，不会引起严重的问题。 结扎本身代表了最关键的一步。左降支冠状动脉很难被可视化，并且常常需要不以约束。因此，一些解剖参考点指出，以帮助外科医生进行正确的连接。对于慢性梗死模型中，结扎被放置在心脏的腹侧的中间，耳廓和顶点之间，主要的前静脉（ 图2B）的结尾的上方。效率得以CONTRO通过可视化的灰色的在灾区的外观LLED。如果梗塞区域出现前，不包括后壁，一个新的缝合线可以放置到第一缝合的左侧。 LAD的主要根源总是可见耳廓18岁以下，因此不会检测这部分出现严重的问题。然而，耳廓呈现出血的主要危险，需要小心处理。 该程序是由存在适当的设备的限制。的通风机和适当的麻醉系统的小动物是昂贵的并且需要连接到该室的气体和通风系统。此外，动物的密切监督是必要的，在第一周后过程，以检测可能的临床。为了检验在试验过程中的心脏功能，高分辨率超声，复杂朗根多夫灌注的系统，或小室内导管测量是必需的，INVolving成本高，更多的专业知识。 考虑到心肌梗死，有可用来重现体外的事件的复杂性，没有可供选择的方法。根据感兴趣的点，分离的心脏中的Langendorff系统的离体灌注提供有关收缩，心脏功能和存活心肌响应于不同的刺激或药物的资料。然而，它不包括血液成分和免疫系统的所有的干扰，并且它不表示心肌梗死后长期研究重塑和愈合。 在进行心肌梗死过程之后，所有其它功能性分析，可以进行，例如脑室压力测量，超声（小动物超声系统）或分离的心脏的Langendorff灌注。此外，所有的生物和分子分析可以被执行，以确定细胞，蛋白质，的mRNA，微小RNA，锗未列名或其他生物标志物，其可以作为治疗目标开发用于心肌梗死新的治疗策略。

心肌梗死是死亡的主要原因工业化国家之一。尽管诊断和治疗方法的不可否认的进展，心血管疾病仍然是死亡的主要原因。鉴于预期提高生活和生命有关的风险，不断增加心血管疾病的发病率，预计在未来。因此，有强烈的需要建立和验证新的方法用于心血管疾病的治疗。人类研究的信息从它的局限性患，这些研究通常是不足以解释和理解的机制在分子水平上，无法提供解决这些严重的健康问题。 此外，基础研究已因限于复杂性和难度重现心血管疾病的机制，在实验室。因此，为了增加约心肺的病理生理我们的知识血管疾病，它是必不可少的，以验证动物模型1,2。然而，为了鉴定参与愈合心肌梗塞，分析在不同时间点后是必要的分子事件的所有级联，导致了大量的动物实验。 心肌梗塞实验通常通过使用动物模型进行的。诱发心肌梗死的小动物是3-11来探讨比大动物模型细胞和分子事件的最合适和最有效的模式。而且，没有其他的物种呈现的转基因或基因敲除品系小鼠12的可用性。这些小鼠模型在其它疾病，包括心血管病症非常有用的（如动脉粥样硬化，在支架内再狭窄）13,14。此外，低怀孕期间和高数后代资格小鼠模型作为最有吸引力的系统来研究心肌INFA的分子机制rction 12。 尽管如此，心脏小鼠的规模预计操纵显微过程中精度高。这样的教学资质和熟练的手术人员是一个耗时和工作的过程。因此，我们在此提出一个详细的显微程序，包括技巧和窍门来引导甚至平均资历的合作者，如学生或技术人员进行小鼠复杂的心肌梗死模型。 最初，插管是由短插管的装置，而无需使用气管切开进行。胸切口位于肋间区域，避免肋或/和周围组织的损伤。该子步骤是高度相关的，以确保快速恢复和愈合15。结扎由差动对慢性缺血和缺血/再灌注模型中，对于高存活率，同时仍保持一个显著梗塞大小。我们的经验表明，塔用丝线缝合吨相比，低温伤害16保证了较高的可重复性。 总之，这里所描述的方法适用于既慢性缺血和缺血/再灌注模型中的小动物。在此过程中提出的技巧和窍门，旨在实现与即使是低或平均合格人员将其应用在小动物模型。

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minimal invasive surgical procedure of inducing myocardial infarction in mice

心肌梗死仍然死在西方国家的主要原因，尽管支架的发展区域，在过去的几十年长足的进步。对于澄清潜在机制和新的治疗策略的发展，有效的动物模型的有效性是强制性的。由于我们在体内条件需要新的见解心血管疾病的病理机制，以打击心肌梗死，动物模型的有效性是一个重要方面。然而，保护动物是在这样的背景下高度相关。因此，我们建立了心肌梗死的小鼠微创和简单的模型，它保证了高重复性和动物的存活率。因此，这种模式满足的3R原则动物实验的要求（更换，细化和还原），并保证所需的cardiov治疗策略进一步发展的科学信息血管性疾病。

心肌梗死过程发生在25-30分钟，并示出了死亡率为10％。手术后，小鼠从麻醉中恢复的下一个15分钟之内。任何物理损伤，观察到鼠标操作。然而，有心脏破裂一个星期后慢性心肌梗死后的风险较高，如果修复过程期间炎症阶段被扰乱。因为心脏是能够泵送期间显著改变其尺寸，重要的是对所有所收集的心要被停止在相同的位置，例如在diastola。这可以通过用饱和氯化钾溶液灌注心脏来实现。增加外K +浓度的块中的离子泵，降低心肌细胞的膜静息电位，造成心脏活动的舒张期停滞。 梗塞面积可以看出，在超声分析（ 图3A，下图）。在比较正常心肌，缺血区出现薄和运动减弱（ 图3A，上图）。根据所使用的模型，该梗塞大小会有所不同。慢性梗死模型诱导的顶点（ 图3B）的圆形，透梗死，而缺血/再灌注诱导的薄的中间壁和在所有心脏（ 图3C）。有许多方法来确定梗死面积。如果目的是要分析对心脏存活的直接作用，伊文思蓝/ TTC染色18被指示要进行再灌注至少2小时后，才能够看到在心肌的任何变化。部分可立即染色后进行分析（ 图3B，中图）或可以保持载玻片之间在福尔马林2-3天（ 图3C，中图）。蓝色区域代表健康心肌，不受缺血。红色区域代表第里面存活心肌ë缺血区（风险心肌），而白色区域表示的死组织。通常，梗塞大小被表示为百分比从风险区。 成熟瘢痕所得重构处理之后可以很容易地通过immunohistolgy使用Gomori法的一步法染色测定。被蓝染梗死和染红健康心室区（ 图3B和C，右图）从每个级别的第一部分，直到二尖瓣确定。为了避免由于LAD的不同层次的结合的变化，从所有部分中的梗塞被认为并表示为总左心室体积的百分比。可以达到15-20％的慢性梗塞模型和10-15％后缺血/再灌注模型中的梗死体积。此外，慢性梗死模型将诱发加重扩张，在缺血/再灌注模型没有观察到（ 图3B和C右击牛逼面板）。 常规染色程序都可以使用，例如：用于揭示血管生成CD31染色（红色， 图4A）或平滑肌肌动蛋白染色来确定肌纤维母细胞（绿色， 图4B）。双荧光染色也可应用，以确定不同的目标分子在梗死区域中，由于不存在心肌没有给出自动免疫荧光（ 图4C）。 <img alt="图1" src="/files/ftp_upload/52197/52197fig1.jpg" /> 图1： 内侧切口和插入气管插管的套管（一）。金属套管穿过组织（B）的透明的stereomicroscopic可视化。的气管环（蓝色箭头）和套管（黑色箭头）被指出。肋间切口慢性心肌梗塞莫德尔和LAD（三）结扎。结扎位于心脏（耳廓和先端，黑色在下部面板之间）的中间，以作为参考的静脉（示意蓝色，下图）的末端。动脉的两个分支应绑定（红色中下图）。灰色的颜色表明梗死面积，它出现在心脏（右下图）的下半部分。结扎的缺血/再灌注模型耳廓下进行，装订LAD的主体（红色中下图）在硅管（右侧）（D）中 。灰色的颜色表示梗死面积，这是目前在整个心脏（右下图）。 <a href="https://www.jove.com/files/ftp_upload/52197/52197fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/52197/52197fig2.jpg" /> 图2： </str翁&gt;肋缝线密封件，如果内侧缝线定位在90°在两个慢性（A）一种角胸切口和缺血/再灌注模型（B，左图）。 在体内成像的肋缝合（C，左面板），肌肉缝合（C，中图）和皮肤缝合（C，右图）。 <a href="https://www.jove.com/files/ftp_upload/52197/52197fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/52197/52197fig3.jpg" /> 图3：超声心动图图像 。普通（A，上图）和梗死区（A，下图）的照片，在长轴采集（纵向，左图）或短轴（横向，右图）..脑梗塞引起的慢性结扎（B）和由1小时缺血再灌注（℃）。伊文思蓝/ TTC染色可以识别灌注（蓝色）/非灌注区域以及可行的（红色）/死（白色）心肌（B，C，中图）。莫里的一步法染色允许梗死区（蓝色）的识别，并从正常区域（红色）（B，C，右图）区分它们。 <a href="https://www.jove.com/files/ftp_upload/52197/52197fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/52197/52197fig4.jpg" /> 图4：不同的染色可以在梗死区中进行，如CD31，以描述新血管生成（A，红色，简单箭头），或平滑肌肌动蛋白为肌成纤维细胞（B，克颖，简单的箭头），以及双重染色（C，CD31-红/平滑肌肌动蛋白绿色），counterstained用DAPI的细胞核（蓝色）。肌纤维母细胞可以很容易地从大或小动脉平滑肌细胞，它们总是伴随着内皮细胞层（C，箭头）进行区分。双箭头指向红细胞自体荧光。规模杆50微米。

Watch this Scientific Journal Video about Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice at JoVE.com

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

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 ...

minimal-invasive-surgical-procedure-inducing-myocardial-infarction

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27.1K 次观看.  RWTH Aachen University, Germany. A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

视频: Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Acute Myocardial Infarction in Rats

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular diseases. Animal models that mimic human cardiac disease, such as myocardial infarction (MI) and ischemia-reperfusion (IR) that induces heart failure as well as pressure-overload (transverse aortic constriction) that induces cardiac hypertrophy and heart failure (Goldman and Tarnavski), are useful models to study cardiovascular disease. In particular, myocardial ischemia (MI) is a leading cause for cardiovascular morbidity and mortality despite controlling certain risk factors such as arteriosclerosis and treatments via surgical intervention (Thygesen). Furthermore, an acute loss of the myocardium following myocardial ischemia (MI) results in increased loading conditions that induces ventricular remodeling of the infarcted border zone and the remote non-infarcted myocardium. Myocyte apoptosis, necrosis and the resultant increased hemodynamic load activate multiple biochemical intracellular signaling that initiates LV dilatation, hypertrophy, ventricular shape distortion, and collagen scar formation. This pathological remodeling and failure to normalize the increased wall stresses results in progressive dilatation, recruitment of the border zone myocardium into the scar, and eventually deterioration in myocardial contractile function (i.e. heart failure). The progression of LV dysfunction and heart failure in rats is similar to that observed in patients who sustain a large myocardial infarction, survive and subsequently develops heart failure (Goldman). The acute myocardial infarction (AMI) model in rats has been used to mimic human cardiovascular disease; specifically used to study cardiac signaling mechanisms associated with heart failure as well as to assess the contribution of therapeutic strategies for the treatment of heart failure. The method described in this report is the rat model of acute myocardial infarction (AMI). This model is also referred to as an acute ischemic cardiomyopathy or ischemia followed by reperfusion (IR); which is induced by an acute 30-minute period of ischemia by ligation of the left anterior descending artery (LAD) followed by reperfusion of the tissue by releasing the LAD ligation (Vasilyev and McConnell). This protocol will focus on assessment of the infarct size and the area-at-risk (AAR) by Evan's blue dye and triphenyl tetrazolium chloride (TTC) following 4-hours of reperfusion; additional comments toward the evaluation of cardiac function and remodeling by modifying the duration of reperfusion, is also presented. Overall, this AMI rat animal model is useful for studying the consequence of a myocardial infarction on cardiac pathophysiological and physiological function.

The rat model of acute myocardial infarction (AMI) is useful to study the consequence of a MI on cardiac pathophysiological and physiological function.

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

The mammalian heart is incapable of significant regeneration following an acute injury such as myocardial infarction1. By contrast, urodele amphibians and teleost fish retain a remarkable capacity for cardiac regeneration with little or no scarring throughout life2,3. It is not known why only some non-mammalian vertebrates can recreate a complete organ from remnant tissues4,5. To understand the molecular and cellular differences between regenerative responses in different species, we need to use similar approaches for inducing acute injuries.
In mammals, the most frequently used model to study cardiac repair has been acute ischemia after a ligation of the coronary artery or tissue destruction after cryoinjury6,7. The cardiac regeneration in newts and zebrafish has been predominantly studied after a partial resection of the ventricular apex2,3. Recently, several groups have established the cryoinjury technique in adult zebrafish8-10. This method has a great potential because it allows a comparative discussion of the results obtained from the mammalian and non-mammalian species.
Here, we present a method to induce a reproducible disc-shaped infarct of the zebrafish ventricle by cryoinjury. This injury model is based on rapid freezing-thawing tissue, which results in massive cell death of about 20% of cardiomyocytes of the ventricular wall. First, a small incision was made through the chest with iridectomy scissors to access the heart. The ventricular wall was directly frozen by applying for 23-25 seconds a stainless steel cryoprobe precooled in liquid nitrogen. To stop the freezing of the heart, fish water at room temperature was dropped on the tip of the cryoprobe. The procedure is well tolerated by animals, with a survival rate of 95%.
To characterize the regenerative process, the hearts were collected and fixed at different days after cryoinjury. Subsequently, the specimen were embedded for cryosectioning. The slides with sections were processed for histological analysis, in situ hybridization and immunofluorescence. This undertaking enhances our understanding of the factors that are required for the regenerative plasticity in the zebrafish, and provide new insights into the machinery of cardiac regeneration. A conceptual and molecular understanding of heart regeneration in zebrafish will impact both developmental biology and regenerative medicine.

Zebrafish represents a valuable model to study the mechanisms of heart regeneration in vertebrates. Here, we present a protocol for induction of a heart infarct in adult zebrafish using cryoinjury. This method results in massive cell death within 20% of the ventricular wall, similar to that observed in mammalian infarcts.

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

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 ideal model for testing novel therapeutic strategies.
In vivo magnetic resonance imaging (MRI) gives excellent views of the heart noninvasively with clear anatomical detail, which can be used for accurate functional assessment. Contrast agents can provide basic measures of tissue viability but these are nonspecific. Positron emission tomography (PET) is a complementary technique that is highly specific for molecular imaging, but lacks the anatomical detail of MRI. Used together, these techniques offer a sensitive, specific and quantitative tool for the assessment of the heart in disease and recovery following treatment.
In this paper we explain how these methods are carried out in mouse models of acute myocardial infarction. The procedures described here were designed for the assessment of putative protective drug treatments. We used MRI to measure systolic function and infarct size with late gadolinium enhancement, and PET with fluorodeoxyglucose (FDG) to assess metabolic function in the infarcted region. The paper focuses on practical aspects such as slice planning, accurate gating, drug delivery, segmentation of images, and multimodal coregistration. The methods presented here achieve good repeatability and accuracy maintaining a high throughput.

We describe how to perform MRI and PET imaging of the mouse heart. The protocol is tailored to assess treatment efficacy in models of myocardial infarction and heart failure.

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

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One important prerequisite is a good understanding of all safety and efficacy aspects obtained in a large animal model that validly reflect the human scenario of myocardial infarction (MI). Pigs are widely used in this regard since their cardiac size, hemodynamics, and coronary anatomy are close to that of humans. Here, we present an effective protocol for using the porcine MI model using a closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. This approach is based on 90 min of myocardial ischemia, inducing large left ventricle infarction of the anterior, septal and inferoseptal walls. Furthermore, we present protocols for various measures of outcome that provide a wide range of information on the heart, such as cardiac systolic and diastolic function, hemodynamics, coronary flow velocity, microvascular resistance, and infarct size. This protocol can be easily tailored to meet study specific requirements for the validation of novel cardioregenerative biologics at different stages (i.e. directly after the acute ischemic insult, in the subacute setting or even in the chronic MI once scar formation has been completed). This model therefore provides a useful translational tool to study MI, subsequent adverse remodeling, and the potential of novel cardioregenerative agents.

This protocol describes the porcine myocardial infarction (MI) model using a 90 min closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. Furthermore, the protocol for several outcome parameters, such as cardiac function, hemodynamics, microvascular resistance, and infarct size, are also presented.

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 ...

Primary Outcome Assessment in a Pig Model of Acute Myocardial Infarction

Mortality after acute myocardial infarction remains substantial and is associated with significant morbidity, like heart failure. Novel therapeutics are therefore required to confine cardiac damage, promote survival and reduce the disease burden of heart failure. Large animal experiments are an essential part in the translational process from experimental to clinical therapies. To optimize clinical translation, robust and representative outcome measures are mandatory. The present manuscript aims to address this need by describing the assessment of three clinically relevant outcome modalities in a pig acute myocardial infarction (AMI) model: infarct size in relation to area at risk (IS/AAR) staining, 3-dimensional transesophageal echocardiography (TEE) and admittance-based pressure-volume (PV) loops. Infarct size is the main determinant driving the transition from AMI to heart failure and can be quantified by IS/AAR staining. Echocardiography is a reliable and robust tool in the assessment of global and regional cardiac function in clinical cardiology. Here, a method for three-dimensional transesophageal echocardiography (3D-TEE) in pigs is provided. Extensive insight into cardiac performance can be obtained by admittance-based pressure-volume (PV) loops, including intrinsic parameters of myocardial function that are pre- and afterload independent. Combined with a clinically feasible experimental study protocol, these outcome measures provide researchers with essential information to determine whether novel therapeutic strategies could yield promising targets for future testing in clinical studies.

Reliable and accurate outcome assessment is the key for translation of preclinical therapies into clinical treatment. The current paper describes how to assess three clinically relevant primary outcome parameters of cardiac performance and damage in a pig acute myocardial infarction model.

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

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 regeneration, and to define new targets for therapeutics. For decades, models of complete heart regeneration existed in amphibians and fish, but a mammalian counterpart was not available. The recent discovery of a postnatal window during which mice possess regenerative capabilities has led to the establishment of a mammalian model of cardiac regeneration. A surgical model of mammalian cardiac regeneration in the neonatal mouse is presented herein. Briefly, postnatal day 1 (P1) mice are anesthetized by isoflurane and placed on an ice pad to induce hypothermia. After the chest is opened, and the left anterior descending coronary artery (LAD) is visualized, a suture is placed around the LAD to inflict myocardial ischemia in the left ventricle. The surgical procedure takes 10-15 min. Visualizing the coronary artery is crucial for accurate suture placement and reproducibility. Myocardial infarction and cardiac dysfunction are confirmed by triphenyl-tetrazolium chloride (TTC) staining and echocardiography, respectively. Complete regeneration 21 days post myocardial infarction is verified by histology. This protocol can be used to as a tool to elucidate mechanisms of mammalian cardiac regeneration after myocardial infarction.

This protocol describes a highly reproducible model of cardiac regeneration by surgical induction of myocardial infarction in the left ventricle of postnatal day 1 mice. The method involves induction of hypothermic anesthesia and ligation of the left anterior descending coronary artery.

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 ...

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

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any potential therapeutic approach directed against acute left ventricular (LV) dysfunction. Current imaging modalities (e.g., echocardiography and magnetic resonance imaging) have several limitations since data on LV pressure cannot directly be measured. LV catheterization in mice undergoing coronary artery occlusion could serve as a novel method for a real-time evaluation of LV function.
At the beginning of the procedure, mice were anesthetized followed by endotracheal intubation. For LV catheterization, the right carotid artery was exposed via middle-neck incision. The catheter was introduced and placed into the LV cavity. Left thoracotomy was conducted and the left main coronary artery (LCA) was ligated. To induce reperfusion, the suture was released after 45 min. Pressure-volume data was recorded at all times.
Ligation of the LCA caused a decrease in LV systolic function as evidenced by a 30% reduction in stroke volume, LV ejection fraction (EF), and cardiac output. Maximum dP/dt as a parameter for LV contractility was also significantly reduced and diastolic function was severely impaired (minimum dP/dt -40%). Reperfusion over a period of 20 min did not lead to a complete recovery of LV function.
Real-time pressure-volume analysis served as a valid procedure for monitoring cardiac function during acute myocardial infarction in mice. Maintaining stable anesthesia and a standardized surgical approach was crucial to ensure valid results. As the early phase of acute myocardial infarction is critical for morbidity and mortality, the delineated method could be beneficial for preclinical evaluation of new strategies for cardioprotection.

Acute myocardial infarction in mice induces acute but incompletely characterized changes in left ventricular (LV) function. LV catheterization in mice undergoing coronary artery occlusion serves as a novel method for a real-time evaluation of LV function.

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 ...

A Cryoinjury Model to Study Myocardial Infarction in the Mouse

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we demonstrate a murine cryoinjury infarct model that generates precise infarct sizes with high reproducibility and replicability. In brief, after intubation and sternotomy of the animal, the heart is lifted from the thorax. The probe of a handheld liquid nitrogen delivery system is applied onto the myocardial wall to induce cryoinjury. Impaired ventricular function and electrical conduction can be monitored with echocardiography or optical mapping. Transmural myocardial remodeling of the infarcted area is characterized by collagen deposition and loss of cardiomyocytes. Compared to other models (e.g., LAD-ligation), this model utilizes a handheld liquid nitrogen delivery system to generate more uniform infarct sizes.

This article demonstrates a model to study cardiac remodeling after myocardial cryoinjury in mice.

研究小鼠心肌梗死的低温损伤模型

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we ...

Semi-Minimal Invasive Method to Induce Myocardial Infarction in Rats and the Assessment of Cardiac Function by an Isolated Working Heart System

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily and highly reproducible MI induction procedure is required to obtain further insight and better understanding of the underlying pathological changes. This procedure can also be used to evaluate the effects or potency of new and promising treatments (as drugs or interventions) in acute MI, subsequent remodeling and heart failure (HF). After intubation and pre-operative preparation of the animal, an anesthetic protocol with isoflurane was performed, and the surgical procedure was conducted&#160;quickly. Using a minimally invasive approach, the left anterior descending artery (LAD) was located and occluded by a ligature. The occlusion can be performed acutely for subsequent reperfusion (ischemia/reperfusion injury). Alternatively, the vessel can be ligated permanently to investigate the development of chronic MI, remodeling or HF. Despite common pitfalls, the drop-out rates are minimal. Various treatments such as remote ischemic conditioning can be examined for their cardioprotective potential pre-, peri- and post-operatively. The post-operative recovery was quick as the anesthesia was precisely controlled and the duration of the operation was short. Post-operative analgesia was administered for three days. The minimally invasive procedure reduces the risk of infection and inflammation. Furthermore, it facilitates rapid recovery. The &#8220;working heart&#8221; measurements were performed ex vivo and enabled precise control of preload, afterload and flow. This procedure requires specific equipment and training for adequate performance. This manuscript provides a detailed step-by-step introduction for conducting these measurements.

This article presents an efficient method to perform myocardial ischemia and subsequent chronic reperfusion in rats using a minimally invasive approach. In addition, left ventricular hemodynamic function of rats is assessed by echocardiography and isolated working heart methods.&#160;

诱导大鼠心肌梗死的半微创方法及独立工作心脏系统对心脏功能的评估

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily ...

A Modified Simple Method for Induction of Myocardial Infarction in Mice

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI remodeling and evaluation of novel therapeutics. Different methods (e.g., isoproterenol treatment, cryoinjury, coronary artery ligation, etc.) have been used to induce MI. Compared with isoproterenol treatment and cryoinjury, coronary artery ligation may better reflect the ischemic response and chronic remodeling after MI. However, traditional methods for coronary ligation in mice are technically challenging. The current study describes a simple and efficient process for induction of MI in mice with readily available materials. The mouse chest skin was cut open under stable anesthesia. The heart was immediately externalized through the intercostal space after blunt separation of the pectoralis major and pectoralis minor. The left anterior descending branch (LAD) was ligated with a 6-0 suture 3 mm from its origin. Following LAD ligation, staining with 2,3,5-Triphenyltetrazolium chloride (TTC) indicated successful induction of MI and temporal changes of post-MI scar size. Meanwhile, survival analysis results showed overt mortality within 7 days after MI, mainly due to cardiac rupture. Moreover, post-MI echocardiographic assessment demonstrated successful induction of contractile dysfunction and ventricular remodeling. Once mastered, an MI model can be established in mice within 2-3 min with readily available materials.

Under adequate anesthesia, the mouse heart was externalized through the intercostal space, and myocardial infarction was successfully induced by ligating the left anterior descending artery (LAD) using materials readily available in most laboratories.

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 ...