This work was supported by Merit Review awards (BX002332 and BX000640) from the Biomedical Laboratory Research and Development Service of the Veterans Affairs Office of Research and Development, National Institutes of Health (R15HL129140), and funds from Institutional Research and Improvement account. The project is supported in part by the National Institutes of Health grant C06RR0306551.

在动物身上进行的所有的外科手术应根据指南的护理实验动物13或其他适当的伦理准则的使用和进行。协议应在继续之前，与相应的机构的动物福利委员会的批准。 1.准备心电图注:在开始之前，不要包括手套，眼镜和一个干净的实验室外套或一次性的礼服个人防护装备。 <ol><li>使用非酒精和非漂白基于除污溶液清洁所述ECG垫。轻轻使用微妙的任务抹布擦拭掉多余的溶液，以确保电极焊盘不被损坏。 </li><li>如果ECG垫具有加热功能，使用它。麻醉小鼠倾向于迅速失温。加热垫40-42℃，在整个手术14保持正常体温的体温为37℃。显示器机身脾气ATURE使用直肠体温计。必要时调整垫温度保持体温〜37℃。 </li><li>由于大部分开胸用鼠标趴在它的后面（平卧）执行，确保切换翻转的"仰卧"设置。许多心电图垫具有的功能俯卧和仰卧位置之间切换。若未选择正确的方向可能会导致心电图事件的不实陈述。 </li><li>麻醉，用5％异氟醚吸入和1升/分钟氧气鼠标。一旦小鼠被麻醉，转印鼠标心电图垫配备有麻醉鼻锥和减少异氟烷以2％和1升/分钟的氧气。确认通过确保鼠标在鼠标的脚用钳子捏不恰当的反应麻醉。 </li><li>敷眼润滑药膏薄薄一层在老鼠的眼睛，以防止干燥和角膜损害，同时麻醉。 </li><li>清洁鼠标的爪子用湿擦拭以去除所有可见的床上用品可能卡住的爪子或可以与电脉冲从爪子到ECG垫的传输造成干扰。干爪子带有擦拭。 </li><li>应用高导电电解质凝胶的少量（比美元硬币稍小），以每个在ECG垫的四个金属电极。 注意:一定要只适用凝胶少量的太多凝胶使得它难以自抑的爪子用胶布垫。此外，爪子很可能在手术过程中滑出克制的，如果他们申请磁带之前湿润。 </li><li>在仰卧位鼠标，可以使用明确医用胶带每个爪子抑制到其相应的电极（ 图1）。首先按每个爪子到一块胶布，然后坚持的磁带心电图垫。确保每个约束爪是与电解质凝胶和电极接触。 </li></ol> 2.获取心电图<ol><li>根据用于心电采集，conf下的设备上igure机器，使得ECG波形可以实时可视化。用于使用超声机器上的生理监测设定心电图记录，一个活B模式图像将具有沿屏幕的底部运行ECG波形。 注意:请参阅个人计算机用户指南，确定如何最好地配置设备。 </li><li>通过按压超声机器上的B模式键或其他ECG记录设备的等效使心电波形的实时可视化。 <ol><li>调整分辨率，占幅度的差异。如果Q波的R波或槽（谷）的峰值都出了视觉帧的，调整分辨率，直到波形的整个高度可被观察到。 注:这可以在生理设置选项卡下的超声心动图检查机器上完成。点击增加或减少箭头，直到整个波形是可见的。 </li></ol></li><li>任何时候，一个图像是要OBTained，清除工具的心电图垫。心电图记录用镊子或手指触摸时鼠标会扰乱波形。确保鼠标仍然是和原始心电图上垫记录任何心电图之前。 </li><li>鼠标进行任何手术切口之前，请使用本机的"记录"或"存储"功能。此图片稍后将作为基准进行比较。 </li></ol> 3，手术过程及心电记录<ol><li>开始之前注射麻醉小鼠具有镇痛（丁丙诺啡，1.5微克，腹腔内）。缺血/再灌注外科手术的细节也可以在其他地方5找到。 </li><li>周围的手术部位去除毛发化学或机械和消毒用优碘溶液的区域。使用手术刀，使平行于食道和气管纵切口。轻轻移动到切口的每一侧的淋巴结，直到薄组织覆盖气管是裸露。使用镊子，轻轻直到气管的白软骨环是可见组织中分离出来。 注:我们通常使用奈尔脱毛露。洗剂被施加于手术部位为约1分钟。奈尔然后彻底用生理盐水或水洗掉。此方法是优选的在我们的实验室，因为高分辨率超声心动图（其可检测毛囊）的手术前和手术后进行。但是，需注意避开敏感区域，如生殖器，然后彻底冲洗掉奈尔以避免潜在的皮肤灼伤。 </li><li>迅速从鼻锥去除老鼠的鼻子和插入通风管插入鼠标的嘴，朝喉咙。当通风管的尖端是通过暴露的颈部区域中可见，对准管与气管的开始。轻轻扭动管一边施加上行压力，直到管滑入气管，可从视觉上确认吨至另一侧hrough半透明气管。 </li><li>确保鼠标在插管过程中保持麻醉。从插管暂停和鼠标返回到鼻锥是否开始蠢蠢欲动。 </li><li>使用字符串的循环，通过循环勾鼠标的两颗门牙，并用胶带将字符串结尾的心电图垫稳鼠标的头部，并确保通风管手术过程中不会移动。快速连接的通风管连接到动物呼吸机，并根据鼠标的重量调整通风设置。到位磁带通风管道。 </li><li>覆盖了鼠标在温生理盐水浸湿纱布暴露气管，以保持组织的干燥。 </li><li>使用沿着胸骨的左侧手术刀的垂直切口。 </li><li>使用镊子，轻轻筋膜层从肌肉层分离。小心切割下面的肌肉层不削减可见血管。 </li><li>使用镊子，抢第三肋并拉动轻轻向上。保持在肋握一只手，用手术剪刀小心切开第三和第四肋骨之间的肋间肌组织。确保肺部没有损坏。 注意:肺将缩回到胸腔几乎立即胸腔由手术切口凿孔之后由于压力梯度的丧失。等到肺部在继续之前收回。 </li><li>使用镊子抓，轻轻环绕心脏心包的薄层分开。 </li><li>插入牵开器或手动使用镊子作为肋拉钩到肋骨移动到一个位置，其中心脏是肋之间可见。 注意:这是使得其结扎的放置期间重叠右下爪移动鼠标的左下爪子常见的做法。这有助于在心脏定位，以使左心耳，或耳廓，是结扎的放置过程中很容易地看到。请注意，有效的心电图W¯¯而左下爪子熄灭电极aveforms不能获得。出于这个原因，建议该缝合结扎穿过心肌组织，但结紧固前通过后爪返回其电极。 </li><li>找到LAD视觉，左耳廓下方。迅速插入7-0丝锥形缝合针到心肌足够深入的LAD下通过，但没有那么深穿透LV腔。通过直到有4厘米左右留在缝合结扎的自由（非针）结束缝合丝拉出缝合结扎。 </li><li>开始绑一个简单的缝合打结。一旦缝合丝的自由端已通过环拉以形成结，暂停。 </li><li>同时持有钳缝合丝的自由和针端插入PE-10管的约1cm部分形成结下方，上盖的心外膜表面。 </li><li>如果鼠标的左爪子被超过，返回爪子其亲每个电极。以便在PE-10管缝合到心脏紧结。释放与鼠标所有物理接触，以允许要被记录的ECG。 </li><li>通过对〜10秒让心电波形循环。目视检查，心电波形和记录波形为"闭塞的时代"。如果T波不振幅1分钟内增加，重新评估结扎的位置。 <ol><li>如果T波振幅也不会增加，无论是从研究丢弃动物或试图纠正结扎位置。 </li></ol></li><li>目视检查心肌的颜色来确认LV缺血变白。 </li><li>如果心电图改变和心肌颜色变化表明缺血，成双结对周围的PE-10管缝合。 </li><li>盖打开胸腔用温生理盐水纱布。 </li><li>记录心电图每5-10分钟的局部缺血期的持续时间。 </li></ol> 4.确认再灌注的使用心电图<ol><li>取出盐水纱布海湾环胸腔和可视化的心脏。 </li><li>用刀片切割缝合丝的PE-10管上面。一旦结扎切断，去除PE-10管的部分，轻轻取出从心肌缝合结扎。 </li><li>释放与鼠标所有物理接触，并允许所述ECG波形〜10秒循环。作为记录波形"再灌注的时间。"继续达到所需的实验时间点直到记录ECG波形每次5-10分钟。 </li><li>根据需要调整变化幅度分辨率。如果T波时不除去在PE-10管材和结扎的改变，再灌注未确认。 <ol><li>如果T波时不去除管子改变，无论是从研究丢弃动物或试图纠正结扎位置。 </li></ol></li><li>目视检查心肌组织回红颜色还确认再灌注。 </li><li>关闭胸腔用5-0缝合肋间同时施加温和的压力，鼠标的胸部驱逐手术过程中已经进入过剩空气丝线缝合。然后缝合肌肉层，最后皮肤。注:施加压力至胸腔可能不足以疏散空气的胸腔中所有小鼠。因此，应采用疏散的注射器和针头的方法，以确保所有的空气已经被排出。 </li><li>从电极旋转吸入麻醉关闭和删除鼠标的爪子之前记录的最后一个心电图。增加氧气到2升/分钟，直到鼠标苏醒保持通风。 </li><li>允许小鼠恢复在恒温控制的环境中， 例如，加热垫或暖培养箱，以避免梗塞变性。根据需要由鼠标鬼脸刻度所指示的治疗小鼠与丁丙诺啡24小时的手术后，然后。 注意:程序再灌注也详细Xu 等人讨论了5。 </li></ol>

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The authors have nothing to disclose.

使用心电图改变为确认心肌缺血再灌注补充的方法，确保咬合结扎的准确位置。结扎的位置精度是减少动物之间的数据变化的关键。在小鼠心脏的LAD是一个困难的动脉可视化。因此，补充的视觉与苍白心电图改变将有助于确保结扎并导致组织损伤的正确位置。 自所述ECG垫提供心脏的非侵入性的观点，可以在研究过程中获得的多个心电图。这可以帮助提供更好的理解期间和手术后发生心脏的变化。关键是要得到基线ECG用于在外科手术后的比较。后期组织坏死，甚至室壁瘤可以通过电生理信号和心电图配置变化的变形观测。这可能会提供见解成心脏衰竭的进展。 超声心动图使用机器测量心电图的优点包括之前或缺血/再灌注手术后的心脏结构和功能参数的同时测量。系统记录心电图的局限性包括购买一个超声心动图机的高昂成本。然而，如果试验需要多天恒定ECG监测，有多种可用于心电图记录包括远程遥测心电图单元与对应，可以进行编程，以记录和分析在不同的时间间隔波形软件的装置。然而，许多心电遥测单元需要植入程序或专门生境。此外，许多变质电极选项包括存在电极夹和针头。通过开胸缺血/再灌注手术是高度侵入性的过程。优点使用心电图垫用超声心动图MAC海因包括手术过程中连接到动物没有电线，没有额外的手术非侵入性程序。然而，调查人员应确定自己的实验室和实验需要的最佳设备。 如前面提到的，在异氟烷麻醉降低心脏速率。此外，异氟醚可以是用K ATP通道的激活心脏保护，从而降低梗塞大小，如已在狗19被发现。小鼠全身麻醉可以使用注射剂来诱导。吸入麻醉并特别适合长时间程序提供更高的安全性。然而，吸入麻醉需要复杂和昂贵的设备，如高精度蒸发器和流量计，具体换气系统，高效清除系统，以防止污染。注射麻醉剂的缺点包括在选择的初始剂量，没有精确地调节延长烷的深度机会难度sthesia，长时间恢复等麻醉的选择必须根据研究20的程序和目标的长度进行调整。 使用心电图为证实小鼠脑缺血/再灌注损伤，有利于提高梗塞的一致性和可重复性的补充方法，同时也开启了通过建立定量的趋势技术的未来应用的可能性。调查人员可能会发现一定的实验组中的类似心电图的配置。例如，经遗传修饰的动物团可以与野生型时手术后表现出非常宽的QT间隔。这翔实的数据，如果将调查使用心肌色彩变化缺血再灌注损伤的唯一证实已经错过。对野生型和转基因小鼠之间进行比较研究，考虑到兰贝斯公约准则也可以是有价值的，特别是对于年龄，性别，致盲和动物21随机化。 总之，心肌缺血/再灌注损伤的补充确认提供多种好处。使用心电图作为补充技术可以帮助手术建立一致性。这可能有助于降低所用动物的数量，同时提供更高质量的数据。它也允许调查监测心脏损伤，组织死亡和/或重塑的非侵入性地随着时间的推移。最后，使用心电图心肌缺血/再灌注的确认提供建立量化电趋势的可能性。

心脏疾病仍然死亡全世界1,2的主要原因。不仅是左心室（LV）的最肌肉室，负责从心脏泵血对整个主体3，它是一个常见的心脏损伤部位心肌梗死后4。左心室组织死亡往往会导致心脏收缩功能衰竭。心脏疾病的动物模型是当务之急生物医学心血管研究的进步。小鼠C57BL / 6株已用于动物模型的流行选择由于它们的快速繁殖的时间，成本低和易于在遗传改变。心脏疾病的研究大多数小鼠模型手术涉及到左冠状动脉的LAD支闭塞。在LAD有时称为左钝缘5,6。法援署提供血液到左室前壁和安特罗侧壁。结扎研究的目的是诱导前梗塞，有时延长INT澳劣质和侧壁区7。 经常使用心肌梗塞研究两个模型包括慢性阻塞心肌梗塞和心肌缺血/再灌注损伤。慢性阻塞是通过手术缝合周围，阻止永久通过LAD血流创建。缺血/再灌注损伤被创建以相同的方式多只用一个短暂的，通常是30-60分钟，局部缺血期。实现短暂缺血，周围LAD咬合缝合关系和一个小的PE-10管，其被平行放置在LAD心脏的心外膜表面上，随后再灌注期，其中管子和封闭缝合被去除，血液是使其再次通过动脉并进入心肌流动。缺血/再灌注手术已被认为是临床相关的因再灌注损伤人体平行梗塞治疗，其中包括舞会的性质PT冠状动脉成形术和动脉支架置入，或冠状动脉旁路。典型地，这些手术期间，在小鼠心脏左心室的缺血是由心肌壁的可见苍白证实。但是，通过简单地不断监测条件下的心电图（ECG）垫进行手术，可在ECG波形观察到明显的变化，从而证实小鼠心肌缺血再灌注损伤。 虽然小鼠心脏在许多方面，包括它的四腔结构类似于人类的心脏，心中也有差异。一个明显的区别是成年小鼠的平均静息心脏速率为600 -每分钟（BPM）700次，而成人人类是60-100〜8,9 BPM。此外，在小鼠复极化波，J和T，常合并与除极QRS波群作出明确的ST段难以辨别10。要复杂electrocardiographicall的过程ý确认心肌缺血，它是T波和用作缺血和心肌梗死损伤在人体中的诊断标志物的ST段的抬高，临床上称为STëlevation 米 yocardial 我 nfarction或STEMI。一个人和鼠的波形之间的主要区别是，S波之后紧接着是J-波直接传送到负T波。期间小鼠急性心肌缺血S波减小的幅度，并随后直接由异常的J-波和倒T波11。 T波似乎没有在小鼠中11来表示复极化的显著一部分。尽管命名和鼠标与人类不同，小鼠心肌缺血再灌注心电图确认是仍然可行和相对简单的。为了简化波形解释的缘故，SJT之间的段被称为ST-segmeNT于此。 发表于2013 STEMI指南推荐小于90分钟12患者门到气球时间。这意味着从患者的冠状动脉闭塞的识别的时间帧，直至动脉重新打开应小于90分钟。跳动的心脏不断工作，因此，具有较高的氧化代谢和氧耗3的较高水平。为了提供这一点，毛细血管的网络是可用于每个肌细胞3。只需要一个心脏跳动数耗尽它的氧气和营养供应。在90分钟的窗口，在一个人的缺血性心脏地区将被封锁接收5400名点附近摇摆心脏跳动的富含氧气的血液的价值。在同样的90分钟的窗口，鼠标将有54000 63000心脏跳动。对鼠缺血/再灌注损伤的实验时间点通常是30分钟和60分钟之间。 发展的重要性ING证实了小鼠模型心肌缺血再灌注的补充方法对心肌缺血/再灌注损伤的研究数据的一致性和可重复性产生深远的影响。目视观察心脏的组织颜色改变目前的做法是不够的作为一个独立的诊断。此外，除去管和缝合之后再灌注不能保证。虽然动脉不再打结，动脉可具有在手术期间持续伤害，并可能成为不可能reperfuse。这将是有益的的心电图变化的记录，以确认再灌注而不是依赖于心肌苍白和发红（红色）的意见。心不显示缺血/再灌注损伤的标记就可以迅速被标记，并就如何继续处理的决定可以由侦查进行。 最后，建立心电图的记录从基线整个日变化Ë缺血再灌注时间允许调查人员继续初次手术后监测心脏。目前调查的手术完成后尽快忘记的心脏。心电图是深入了解手术后在心肌小时发生至数天变化的简单方法。心电图记录的时间点，手术后可以揭示表明持续或恶化的组织死亡后发Q波。然而，为了有效地测厚仪新的或恶化的心电图标记，一个基线ECG必须可用于比较。 该协议将展示如何准备，获取和解读心电图，确认使用8小鼠心脏缺血再灌注 - 19周龄雄性C57BL / 6小鼠。

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confirmation of myocardial ischemia and reperfusion injury in mice using surface pad electrocardiography

Many animal models have been established for the study of myocardial remodeling and heart failure due to its status as the number one cause of mortality worldwide. In humans, a pathologic occlusion forms in a coronary artery and reperfusion of that occluded artery is considered essential to maintain viability of the myocardium at risk. Although essential for myocardial recovery, reperfusion of the ischemic myocardium creates its own tissue injury. The physiologic response and healing of an ischemia/reperfusion injury is different from a chronic occlusion injury. Myocardial ischemia/reperfusion injury is gaining recognition as a clinically relevant model for myocardial infarction studies. For this reason, parallel animal models of ischemia/reperfusion are vital in advancing the knowledge base regarding myocardial injury. Typically, ischemia of the mouse heart after left anterior descending (LAD) coronary artery occlusion is confirmed by visible pallor of the myocardium below the occlusion (ligature). However, this offers only a subjective way of confirming correct or consistent ligature placement, as there are multiple major arteries that could cause pallor in different myocardial regions. A method of recording electrocardiographic changes to assess correct ligature placement and resultant ischemia as well as reperfusion, to supplement observed myocardial pallor, would help yield consistent infarct sizes in mouse models. In turn, this would help decrease the number of mice used. Additionally, electrocardiographic changes can continue to be recorded non-invasively in a time-dependent fashion after the surgery. This article will demonstrate a method of electrocardiographically confirming myocardial ischemia and reperfusion in real time.

正常鼠心电图与用于电事件P，Q，R，S，J和T P字母标记显示在图2中是初始心房去极化。 QRS是去极化在心室的浪潮。 J是早期复极T表示异构的复极又称恢复11。应该指出的是，许多实验室不使用的J形波命名法，而是指SJT段作为ST段10,15-17。在这里，结果和分析具有代表性和基于关闭40只小鼠的实验观察。大多数小鼠在手术的过程中表现出类似的波形级数。未表现出相似的波形小鼠标记进行进一步的分析，被认为非梗塞的动物。类似波形的结果也有报道通过Jong 等15。 鼠心苏ffering从局部缺血因结扎通常显示增加了R波振幅还有JT-段其次是ST段的最终抬高的超急性调峰图3显示了急性心肌缺血的第一个迹象。 T波的超急性峰值。如在该图中可以看出，T波已在振幅从基线情况增加。然而，这是没有ST段抬高，因为S波仍突出深和带负因为它在基线波形。 心电图基线配置显示负投影S波（ 图2）。随着时间的推移，心电监护，注意变化。 ST段定义为S波的结束和T波的起点之间的线段。这种ST段是人类清楚。由于心脏高的速度，这个段在小鼠中合并和一个额外的，早期复极化"J-波"SEParates S-和T波。因此，该S波至等电线或更高的高度应视为ST段抬高的鼠版本。在图4中局部缺血的早期梗塞的进展可通过ST段抬高看到。这里的S波被显示在升高的幅度，等电位线的上方。歼波也升高，特别是当相对于基准波形（ 图2）。因此，ST段升高其表示伤害/梗死10。 图5所示一只小鼠的从基线的进程通过灌注一路。第一波形显示正常窦性心律这是所记录的基线。第二个波形显示的心电图1分钟结扎被捆绑和动脉闭塞开始后。在这条线的红色圆圈表示超急性T波峰值。如果比较的Wi日"基线"，很显然，T波升高。第三波形示出了在5分钟的时间点的完整的ST段抬高。在"1分钟缺血"的形象，S波仍然突出负，传递等电位线。然而，在5分钟的复杂的S-部分不尽可能负，因为它应该前进到J-和T波前到达。因为S-和T波之间的段是从等电位线升高此被描述为ST段抬高。局部缺血的另一种电生理标记加宽从QRS波群的开始延伸，并继续进行，直到T波的结束的QT间期16。在缺血20分钟时，QT间期已经扩大和ST段仍在升高。缺血45分钟后，QT间期遗存扩大和ST段持续升高。 心肌再灌注有过性脑缺血30分钟或更小应导致心电图在小鼠返回回基线条件。普雷达和Burlacu建立鼠心电图变化，缺血时间，和梗死严重性17之间的相关性。据观察，在30分钟时间缺血没有造成永久性的心电图变化，而1小时的缺血时间并造成永久性的心电图改变。此外，缺血后24小时闭塞动脉缺血再灌注没有打捞效果17。通常情况下，Q波可以被确定为稍微向下的投影之前的去极化QRS波群。显著，或病理性Q波可以根据相当死肌开始于10设置心肌缺血发作后不久发展。显著Q波被定义为相应的R波的高度的至少1/3或可以通过细长的时间，从而导致在宽Q波。从死心肌偏转ELECTRICA的区域的显著Q波的结果升电流从电极7路程。 5分钟再灌注，深证据后，显著Q波开始出现（ 图5）。此外，T波返回到等电位线（ 图5）。再灌注30分钟后，将负Q波保持，并有可能显示出永久性的损害。在这个时间点，在Q-波宽深，并指出濒死心脏组织周围偏转受损区域（ 图5）电流。 持续缺血后，发展到伤害和梗死导致增强负T波投影（ 图5，30分钟再灌注，第二红圈）。这种增强的T波预测，由于真正的梗死通常是永久性的7。在30分钟再灌注波形第二红色圆圈显示似乎是一个倒置的T波（ 图5）。如果5次30分钟的T波相比，很显然，T波是更突出的消极。这一点，再加上显著Q波为这个心脏永久性的组织损伤的证据。应当注意的是，吸入异氟烷麻醉降低心脏速率，并因此增加的QT间隔。不过，恢复T波振幅不受影响18。 上述变化可以定量分析在电压方面， 图6示出了使出口生理数据作为.csv文件将提供非常大的数据量。除了在馏分-的毫秒价格，提供心电图的值（振幅）可以有选择，如果需要的话，以包括从呼吸，温度探测器，血压袖带等的其他数据。这些量化的数据可被绘制， 如图7。从P波向P波作图一系列波形有助于可视化的ECG配置的趋势。 500毫秒的时间段是一个很好的时间帧来可视化，因为任何更少的时间可能不会导致足够波形和任何额外的时间会使图形出现一个标准计算机显示器上观看时凌乱和电生理事件可能被遗漏或难以识别。 <img alt="图1" src="/files/ftp_upload/54814/54814fig1.jpg" /> 图1:心电图垫鼠标的位置是否正确这款鼠标定位在仰卧位。老鼠的爪子每个人都贴在ECG面垫相应的电极。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54814/54814fig1large.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/54814/54814fig2.jpg" /> 图2: 正常小鼠基线心电波形，正常小鼠心电图基线都标有其用来描述在心脏电活动的字母P，Q，R，S，J和T。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54814/54814fig2large.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/54814/54814fig3.jpg" /> 图3:T波高度也被称为超急性T波或峰值。 T波被放大，比基准T波（图2）更高。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54814/54814fig3large.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/54814/54814fig4.jpg" /> 图4:ST段抬高这个数字显示终端可以观察到的ST段AYS ST段抬高比等电点越高。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54814/54814fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/54814/54814fig5.jpg" /> 图5: 典型的心电图改变过的缺血/再灌注手术的课程这个数字如下一个鼠标在心肌缺血/再灌注术的持续时间。第一波形显示正常窦性心律这是录制的基准（基准）。第二个波形（1分钟缺血）显示波形1分钟结扎被捆绑和动脉闭塞开始后。在这条线的红色圆圈显示超急性T波峰值。在第三波形（5分钟缺血）的红圈显示完整的ST段抬高。在缶的红圈<html> RTH波形（20分钟缺血）显示加宽QT间期和S波仍在升高。在第五波的红色圆圈（45分钟缺血）显示加宽QT段和高架ST段。第六波形（1分钟再灌注）显示第七波形（5分钟再灌注）显示深，显著Q波的形式没有变化显著与45分钟ischemia.The红圈。在第八波形（30分钟再灌注）第一个红色圆圈显示显著Q波，而第二个红圈显示可能的T波增强。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54814/54814fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图6" src="/files/ftp_upload/54814/54814fig6.jpg" /> 图6: 生理数据，此图显示的生理数据，因为它是导出为.csv文件到电子表格。REF ="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54814/54814fig6large.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 <img alt="图7" src="/files/ftp_upload/54814/54814fig7.jpg" /> 图7:绘制毫伏的值在该图中显示的曲线图显示了使用生理数据文件（图6）从连续的，完整的三个波形毫伏值。该图是用点从生理数据文件的简单线图。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54814/54814fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Confirmation of Myocardial Ischemia and Reperfusion Injury in Mice Using Surface Pad Electrocardiography at JoVE.com

Confirmation of Myocardial Ischemia and Reperfusion Injury in Mice Using Surface Pad Electrocardiography

During murine myocardial ischemia/reperfusion surgery, correct placement of the occluding ligature is typically confirmed by visible observation of myocardial pallor. Herein, a method of electrocardiographically confirming ischemia and reperfusion, to supplement observed myocardial pallor, is demonstrated in male C57Bl/6 mice.

小鼠使用表面垫心电图心肌缺血再灌注损伤的确认

Many animal models have been established for the study of myocardial remodeling and heart failure due to its status as the number one cause of mortality ...

confirmation-myocardial-ischemia-reperfusion-injury-mice-using

Nanyang Technological University

Research

JoVE Journal

Medicine

13.2K 次观看.  East Tennessee State University. During murine myocardial ischemia/reperfusion surgery, correct placement of the occluding ligature is typically confirmed by visible observation of myocardial pallor. Herein, a method of electrocardiographically confirming ischemia and reperfusion, to supplement observed myocardial pallor, is demonstrated in male C57Bl/6 mice.

视频: Confirmation of Myocardial Ischemia and Reperfusion Injury in Mice Using Surface Pad Electrocardiography

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

常温心脏骤停和心肺复苏术:缺血再灌注损伤的小鼠模型

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

科研

医学

A Murine Closed-chest Model of Myocardial Ischemia and Reperfusion

Surgical trauma by thoracotomy in open-chest models of coronary ligation induces an immune response which modifies different mechanisms involved in ischemia and reperfusion. Immune response includes cytokine expression and release or secretion of endogenous ligands of innate immune receptors. Activation of innate immunity can potentially modulate infarct size. We have modified an existing murine closed-chest model using hanging weights which could be useful for studying myocardial pre- and postconditioning and the role of innate immunity in myocardial ischemia and reperfusion. This model allows animals to recover from surgical trauma before onset of myocardial ischemia.
Volatile anesthetics have been intensely studied and their preconditioning effect for the ischemic heart is well known. However, this protective effect precludes its use in open chest models of coronary artery ligation. Thus, another advantage could be the use of the well controllable volatile anesthetics for instrumentation in a chronic closed-chest model, since their preconditioning effect lasts up to 72 hours. Chronic heart diseases with intermittent ischemia and multiple hit models are other possible applications of this model.
For the chronic closed-chest model, intubated and ventilated mice undergo a lateral blunt thoracotomy via the 4th intercostal space. Following identification of the left anterior descending a ligature is passed underneath the vessel and both suture ends are threaded through an occluder. Then, both suture ends are passed through the chest wall, knotted to form a loop and left in the subcutaneous tissue. After chest closure and recovery for 5 days, mice are anesthetized again, chest skin is reopened and hanging weights are hooked up to the loop under ECG control.
At the end of the ischemia/reperfusion protocol, hearts can be stained with TTC for infarct size assessment or undergo perfusion fixation to allow morphometric studies in addition to histology and immunohistochemistry.

Surgical trauma induces an inflammatory response. Cytokines and endogenous ligands are known to modulate myocardial infarct size following ischemia and reperfusion. We present a modified closed-chest model of murine ischemia and reperfusion using hanging weights to minimize effects of thoracotomy.

闭胸鼠心肌缺血再灌注模型

Surgical trauma by thoracotomy in open-chest models of coronary ligation induces an immune response which modifies different mechanisms involved in ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

急性肾损伤和伤后纤维化小鼠缺血再灌注模型

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

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

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms at work during MI and developing effective therapeutic approaches requires methodology to reproducibly simulate the clinical incidence and reflect the pathophysiological changes associated with MI. Here, we describe a surgical method to induce MI in mouse models that can be used for short-term ischemia-reperfusion (I/R) injury as well as permanent ligation. The major advantage of this method is to facilitate location of the left anterior descending artery (LAD) to allow for accurate ligation of this artery to induce ischemia in the left ventricle of the mouse heart. Accurate positioning of the ligature on the LAD increases reproducibility of infarct size and thus produces more reliable results. Greater precision in placement of the ligature will improve the standard surgical approaches to simulate MI in mice, thus reducing the number of experimental animals necessary for statistically relevant studies and improving our understanding of the mechanisms producing cardiac dysfunction following MI. This mouse model of MI is also useful for the preclinical testing of treatments targeting myocardial damage following MI.

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

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 ...

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for myocardial infarction and cardiac complications of thoracic surgery. The ability of certain drugs to precondition the myocardium against ischemia reperfusion injury has led to multiple clinical trials, with little success. The isolated heart model allows acute observation of the functional effects of ischemia reperfusion injury in real time, including the effects of various pharmacological interventions administered at any time-point before or within the ischemia-reperfusion injury window. Since brief periods of ischemia can precondition the heart against ischemic injury, in situ aortic cannulation is performed to allow for functional assessment of non-preconditioned myocardium. A saline filled balloon is placed into the left ventricle to allow for real-time measurement of pressure generation. Ischemic injury is simulated by the cessation of perfusion buffer flow, followed by reperfusion. The duration of both ischemia and reperfusion can be modulated to examine biochemical events at any given time-point. Although the Langendorff isolated heart model does not allow for the consideration of systemic events affecting ischemia and reperfusion, it is an excellent model for the examination of acute functional and biochemical events within the window of ischemia reperfusion injury as well as the effect of pharmacological intervention on cardiac pre- and postconditioning. The goal of this protocol is to demonstrate how to perform in situ aortic cannulation and heart excision followed by ischemia/reperfusion injury in the Langendorff model.

The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

诱导和缺血再灌注损伤评估中的Langendorff灌注鼠心

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

肠缺血再灌注损伤的小鼠模型

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

大鼠心肌缺血再灌注后冠状动脉流动储备评价

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

重建显微外科缺血再灌注损伤研究的临床前大鼠模型

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, cardiac stem cell therapy is studied by delivering SCs concurrently with the induction of myocardial injury. However, this approach presents two significant limitations: the early hostile pro-inflammatory ischemic environment may affect the survival of transplanted SCs, and it does not represent the subacute infarction scenario where SCs will likely be used. Here we describe a two-part series of surgical procedures for the induction of ischemia-reperfusion injury and delivery of mesenchymal stem cells (MSCs). This method of stem cell administration may allow for the longer viability and retention around damaged tissue by circumventing the initial immune response. A model of ischemia reperfusion injury was induced in mice accompanied by the delivery of mesenchymal stem cells (3.0 x 105), stably expressing the reporter gene firefly luciferase under the constitutively expressed CMV promoter, intramyocardially 7 days later. The animals were imaged via ultrasound and bioluminescent imaging for confirmation of injury and injection of cells, respectively. Importantly, there was no added complication rate when performing this two-procedure approach for SC delivery. This method of stem cell administration, collectively with the utilization of state-of-the-art reporter genes, may allow for the in vivo study of viability and retention of transplanted SCs in a situation of chronic ischemia commonly seen clinically, while also circumventing the initial pro-inflammatory response. In summary, we established a protocol for the delayed delivery of stem cells into the myocardium, which can be used as a potential new approach in promoting regeneration of the damaged tissue.

Stems cells are continuously investigated as potential treatments for individuals with myocardial damage, however, their decreased viability and retention within injured tissue can impact their long-term efficacy. In this manuscript we describe an alternative method for stem cell delivery in a murine model of ischemia reperfusion injury.

在穆林模型中缺血再灌注损伤后干细胞的心肌内传递延迟

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

器官缺血-输液损伤模拟大鼠肝脏移植模型的血动力学变化

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...