研究报告本出版物中被关节炎，肌肉骨骼国家研究所和皮肤疾病，美国国立卫生研究院的一部分，根据奖号码R01-AR063084支持。内容完全是作者的责任，并不一定代表美国国立卫生研究院的官方意见。

这种动物议定书批准，并按照规定的机构动物护理和使用委员会（IACUC）在俄亥俄州立大学的准则和法规。由当地IACUC制定的所有政策都符合了实验动物福利办公室在美国国立卫生研究院研制的动物实验指南。 1，麻醉和气管插管<ol><li>使用高压灭菌前所有仪器和手术用品。整个手术过程中戴无菌，一次性使用手术手套。保持无菌区域在整个手术过程。使用无菌悬垂的建议，但在视频中没有显示，以便更好地在鼠​​标可视化的解剖标志。 </li><li>单独地将每个小鼠在感应腔和用5％异氟烷和氧气为0.4升/分钟的流速，直到正向反射丧失提供麻醉，然后保持第ë动物用2％异氟醚在100％氧气为0.4升/分钟通过鼻锥管连接到麻醉机，直到气管导管到位的方式流动。用异氟醚麻醉机应适当通风，并配备了木炭过滤器，在操作过程中尽量减少医生暴露于异氟醚烟雾。鼻锥注意到但在视频中未示出，以允许操作的可视插管鼠标。 </li><li>剃除动物的胸带在不同位置的动物毛发推剪比手术平台，以避免手术位置的污染。 </li><li>鼠标放置在手术平台，为后续插管仰卧位。一个简单的小的聚苯乙烯泡沫平台可以作为一个操作平台。盖上预先消毒盖布，以提供无菌表面的平台。放置一个加热垫的平台和悬垂性之间维持小鼠中surgica体温升过程。 </li><li>附加的至少10cm 2-0丝线缝合的长度与带的平台，然后环缝线围绕前上门牙。在靠近（2-3厘米）的锥形定位到平台的在鼠标的鼻部的边缘。拖动鼠标拉紧并用一块胶布尾巴将其固定到平台上。 </li><li>固定腿向身体的两侧的带合股线。重要的是，前面的四肢不过度紧张，因为这可能会危及呼吸。 </li><li>准备剃光手术部位用优碘和酒精的颈部和胸部切口才制成。 </li><li>放置平台，用鼠标头指向操作者的方向。切具有0.5cm正中颈部皮肤切口。分离甲状腺叶在其峡部，以暴露胸骨舌骨肌肌肉，其中气管可以根据肌肉可以看出。 </li><li>除去18号套管针的内针，因此它可以被用来作为intub通报BULLETIN管。针尖可以作为支架，1厘米外管的可作为气管套管。 </li><li>按住鼠标用弯钳的舌头，另一只手稍微动一下向上。通过子宫颈皮肤切口看到气管。用另一只手轻轻地插入插管直至管子被认为是在气管内。 </li><li>只要管子在气管中，将在朝向管另一方面弯曲的镊子，并迅速取出内针。如果管不能被插入到气管，该管应拉出，以避免产生呼吸问题。它指向的管向上的尖端，当它靠近喉部，以避免在插入管进入食道而不是气管是很重要的。 </li></ol> 2，通风和固定<ol><li>提供人工通气与动物呼吸器通气2％异氟烷在氧气中为0.4升/分钟的流速。使用改性Y-SHAPE连接到插管与呼吸机连接。气管导管的正确定位可以通过判断对称的胸部扩张得到证实。 </li><li>设定潮气量为260微升/冲程和通气速率是每分钟130冲程，其可以被调整到体重特定鼠标，如果必要的。 </li><li>取出磁带的尾巴上，然后转动鼠标轻轻将它放在一个正确的卧位为后续手术。用胶带重新固定尾巴和腿的平台。 </li><li>插入直肠探头监测体温并调整变暖垫，保持约37ºC的温度</li><li>用胶带固定探头的平台。布比卡因注射在皮下切口部位麻木的区域切口前作出。 </li></ol> 3。开胸<ol><li>做一个斜切口也就是大约1厘米长在站台2 mm处从左侧在哪里左前腿符合主体（以下，其中腿和身体加入约1-2毫米）的方向胸骨。浅表胸静脉靠近这个位点和切口应使得切口的侧端上升到，但不会切入，静脉。 </li><li>虽然切胸肌肉下方露出肋骨。在此步骤期间避免血管意外伤害。如果出血确实发生，用棉花涂药停止任何出血继续下一步骤7之前。 </li><li>可视化的肋骨和透过薄薄的半透明胸壁肺膨胀。用手术剪，以6-8毫米的切口在第三肋间打开胸腔。这个切口应该是一个最小为2 mm从胸骨处所在的胸廓内动脉的位置。损坏动脉会产生大量出血是难以控制的。 </li><li>将预先消毒自制的胸部拉钩INTØ切口，轻轻地拉了回来，打开切口，使其约8-10毫米宽的同时小心避开肺。该卷收器应附到外科手术平台与标签了。 </li><li>在这一点上的心脏应该是可见的，然而，肺仍然覆盖心脏上的一个部分。轻轻拿起心包弯钳，拉它拆开，然后滑动牵引器背后的组织。在这个操纵肺会抬起并远离心脏。 </li></ol> 4。定位法援署<ol><li>通过解剖镜下找到心脏表面上的法援署。法援署流下心脏壁从靠近心脏向下穿过左心室心尖部的中间。法援署会出现鲜红色，并坚决脉冲。这里的静脉有时被误认为法援署，但是适当的照明可以帮助区分两艘船只。如果光线太亮可能很难体会到的颜色血管之间的差异。 </li><li>用无菌棉球片段的直径为约1-2毫米，以准备在LAD用于连接。将左心房和左心室之间的棉花，这将解除左心房，并帮助揭露LAD和澄清其立场。如果在LAD不能定位，该片段可以进一步使左心房被提升更高，揭示其中LAD来源于主动脉滑动。 </li></ol> 5，LAD结扎<ol><li>对于结扎的理想定位是比左心耳的前端下大约为2毫米。肺动脉干，可以用作标记，以帮助识别左外耳。可替换地，结扎位置可以看作一个点1-2毫米远离左回旋的分支。用弯钳在紧随拟结扎点以下站点轻轻地施加压力。这将使它更容易看到的动脉也将有助于保持心脏的地方并简化绑绷带。用钳子超过5秒钟的时间，不要施加压力，并避免可能改变泵的心脏压缩。 </li><li>使用锥形针通过6-0丝线缝合LAD下方同时用解剖显微镜观察。将针头准确动脉下仿佛置身太深针将进入左心室腔或损坏法援署如果针太浅。如果在LAD受伤取出针和缝合LAD控制出血，但如果出血不能被控制，优选安乐死的动物。 </li><li>使一个松散的双结的缝合，留下通过一个2-3毫米长片的PE-10管放置8 2-3毫米直径的环。 </li><li>拧紧动脉和管道周围循环再固定循环通过把一个额外的活结，小心不要损坏室壁。对于永久性结扎，直接将法援署与结9。通过检查在应该出现结扎后的几秒钟内，该LV的前壁外观苍白色的确认LAD闭塞。 </li><li>取下牵引器并暂时通过与斗牛犬一起夹捏肌肤关闭伤口。的时间缺血保持长度取决于实验设计，但常常是20，30，45或60分钟。鼠标仍然在呼吸机上的LAD动脉阻塞的持续时间。 </li></ol> 6。再灌注<ol><li>缺血期后取出钢夹，然后插入胸部拉钩暴露结扎。解开，取下PE-10管。通过观察返回的LV后15-20秒前壁的粉红色的确认再灌注。 </li><li>离开缝合到位，如果2％三苯基氯化四唑（TTC）和蓝染色将灌注后进行。如果染色是没有必要的，缝合线可以bE法除去。 </li><li>再灌注时间将取决于实验设计中，通常从1小时跨越到24小时。 </li></ol> 7，胸部闭合和术后护理<ol><li>缝制关闭胸腔关闭切口在第三肋间用4-0丝线缝合。重要的是，肺部有明显的缝线，并不会成为被困的3 和第 4肋骨缝合在一起。同时追平了缝线结是有帮助的轻微压力施加到胸前持针器，以尽量减少任何室内空气可能被截留在胸腔。 </li><li>收肌的所有层使用4-0丝线连续缝合。使用尼龙缝合关闭了连续缝合皮肤。可选地，皮肤可以被关闭以间断缝合。 </li><li>当缝合完成后辍异氟醚的流动，而氧气继续流动。当鼠标移动胡须或尾部吧，建议立即进行删除D开始进行尝试自主呼吸。从与插管呼吸​​机取下鼠标仍保留在气管。 </li><li>仔细观察动物，直到鼠标恢复正常的呼吸模式，然后拔管鼠标。该管应缓慢取出，以避免口腔分泌物误吸。 </li><li>确认鼠标是不是在任何呼吸窘迫由它返回一个笼子前，观察它的另一个3-5分钟。如果脱水迹象手术后的观察，提供高达0.5毫升无菌生理盐水腹腔注射。 </li><li>对于手术后的镇痛，管理阿片类镇痛药（丁丙诺啡，0.1毫克/千克），皮下（SC）的前动物门诊，然后提供一个额外的剂量每4-6小时在接下来的24小时。检查窘迫的属相在12小时后手术。使用模拟手术存活心肌梗死，需要痛苦和困扰下从苏复苏的评估rgery。目前公认的最佳做法是提供镇痛前24小时以下的侵入性程序给定的保证，由于体重减轻或疼痛的迹象追加剂量。对于永久性结扎，体重应每天跟踪，以帮助衡量动物的恢复。 </li><li>布洛芬（布洛芬），非类固醇消炎药（NSAID），具有抗炎，镇痛和解热的活性，或其他NSAID，可在动物的饮用水提供为0.2毫克/毫升的溶液用于手术前两天和高达手术后7天随着丁丙诺啡管理任何额外的疼痛/不适。 </li></ol>心肌梗死面积8。测量<ol><li>麻醉和插管鼠标在所需的再灌注时间的尽头。切开胸部皮肤在中线至剑突。打开下方的肋骨和锁骨中线两侧的腹部和隔膜。 </li> &lt;LI&gt;暴露心脏，然后重新结扎在同一位置的法援署。导管插入主动脉所以10％的酞菁蓝，可以慢慢地直接注入主动脉从非缺血区10弄脏的心脏的缺血区的划定。 </li><li>迅速摘除心脏并在30毫米氯化钾（氯化钾溶液）清洗它停止了心脏跳动，并允许更一致的切片。冻结心脏，至少4小时，在-20°C和切心脏成使用心脏矩阵切片装置11 1 mm层厚。 </li><li>培养心脏切片用2％TTC，在37℃40分钟。梗死区被划为白色区域，而活组织染色红。 </li><li>固定染色切片用10％甲醛过夜，这将有助于增加梗死区和正常组织之间的对比度。拍摄切片并计算以危险区（AAR），非缺血区和使用ImageJ软件的梗塞面积。 </li></ol> &lt;P类=“j​​ove_title”&gt; 9。的心脏酶水平测量测量肌钙蛋白小鼠通过获得来自门静脉的血，然后通过离心分离血清，血清中I（cTnI水平）的水平。血清cTnI的水平然后用定量的快速cTnI检测12确定。

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诺亚魏斯勒德博士是创始人兼首席科学​​官TRIM-edicine公司

小鼠心肌缺血再灌注模型是一种有效的方法心血管研究模拟临床急性或慢性心脏疾病13,14。显著的努力已经被应用到发展和完善产生缺血性事件再灌注损伤几种不同类型的动物的心外科手术的方法。虽然有特定的优势，以使用不同的动物系统，鼠标具有导致在小鼠心脏产生心肌I / R广泛关注的特点。其中一个重要的原因是鼠标系统的遗传可追踪性。广泛的选择提供转基因动物，并通过该新?...

心肌梗死（MI）的动物模型是很重要的缺血性心脏疾病1的复杂病理生理机制的研究。缺血 - 再灌注（I / R）损伤是一个重大的贡献MI期间产生的心肌损伤。经冠脉循环的阻塞产生的初始缺血损伤可在心肌梗死患者通过使用血管成形术来及时恢复灌注最小化。尽管这种干预已经死亡的人数大大减少因急性心肌梗死，血流恢复到缺血区会导致I / R损伤，导致心肌细胞的死亡。这种损失心肌质量有助于心输出量和对心脏衰竭恶化下降。因此，这导致从I / R损伤心肌细胞死亡机制的研究是调查在心血管研究的重要防线。外科冠脉结扎术是一种有用的实验技术诱导心肌梗死在各种动物模型的类型，includi纳克的大鼠，狗和猪。在不同的实验室的出版物纷纷出台关于建立I / R损伤2,3小鼠心脏模型的各种方法。为了深入了解这些机制，我们必须获得可靠的动物模型，可以重现心肌梗死病理的几个方面。这种模型的发展也是用于测试治疗方法治疗心肌梗死和相关的I / R损伤至关重要。 大部分的现有的外科手术技术来模拟MI在实验动物涉及手术解剖入胸腔，以暴露左前降支（LAD），然后由一个连字为定义的周期中的时间来产生的缺血性事件闭塞。然后该绑带可以移除，以允许缺血区的I / R损伤和产生再灌注损伤。这些方法中，关于在LAD文献的位置并不总是精确地再现，其中一个主要的限制可以导致变异的心肌梗死的诱发这种做法的严重性。大多数现有技术在心脏前壁一般只描述法援署的大致位置。由于法援署的分支和方向可以在动物个体而异的位置并不总是固定的，可以很容易混淆4,5，手术6时导致潜在的并发症。放置不当的绑带的后果可以从变异诱发左心室完全损害模型的特异性梗塞的大小运行。这里，我们提出对心肌I / R和在小鼠体内的永久结扎，其允许在LAD结扎线放置的改进的准确性的改进方法。通过施加初始切口和内部夹层，以及使用的操作的解除心房，以便更好地欣赏LAD和站点如果出现从主动脉的具体方法。建立在LAD和它的起源位置提供了机会，结扎LAD可重复的方式。心肌I / R和永久结扎该模型不仅降低手术后的变化在梗塞面积，它也可以在操作过程中减少过度出血的发生率。

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a murine model of myocardial ischemia-reperfusion injury through ligation of the left anterior descending artery

急性或慢性心肌梗死（MI）是导致高发病率和死亡率心血管事件。建立病理机制在工作期间心肌梗死并制定有效的治疗方法，需要的方法来重复地模拟临床发病率，并反映与心肌梗死相关的病理生理改变。在这里，我们描述了一种手术方法来诱导心肌梗塞的小鼠模型，可用于短期缺血 - 再灌注（I / R）损伤以及永久性结扎。这种方法的主要优点是便于左前降支（LAD）的位置，以允许该动脉的准确结扎来诱导局部缺血的小鼠心脏的左心室。结扎对LAD准确定位增加梗死面积的可重复性，从而产生更可靠的结果。在结扎位置更高的精确度将提高标准的手术方法来模拟心肌梗死小鼠中，THUS减少了必要的实验动物的数量统计上相关的研究和改进我们的生产心功能不全以下心肌梗死的机制的理解。这种小鼠心肌梗死模型也是靶向心肌损害以下MI治疗的临床前测试非常有用。

在24小时再灌注损伤，心肌梗死面积和地区高危（AAR），用酞菁蓝染料和三苯四氮唑（TTC），法援署结扎可以通过观察心肌组织远端热烫的缝合证实的分析以及前壁的功能障碍。再灌注可以通过红色的回归心肌组织和前壁运动的一些复苏的演示验证。 梗塞区域（白色）应该是区别于领域的风险（红色）和面积没有风险（蓝色）。的酞菁蓝染（ 图1A）应用程序允许?...

Watch this Scientific Journal Video about A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery at JoVE.com

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

我们介绍一种手术方法来诱导实验性缺血/再灌注（I / R）损伤模拟心肌梗死（MI）的小鼠模型，允许在结扎定位左前降支（LAD）上更清晰，增加了重复性MI的实验小鼠。

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 ...

a-murine-model-myocardial-ischemia-reperfusion-injury-through

Research

JoVE Journal

Medicine

38.9K 次观看.  The Ohio State University. 我们介绍一种手术方法来诱导实验性缺血/再灌注（I / R）损伤模拟心肌梗死（MI）的小鼠模型，允许在结扎定位左前降支（LAD）上更清晰，增加了重复性MI的实验小鼠。 

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

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

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 is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

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 ...

科研

医学

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, ...

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, diabetic retinopathy, and retinal vascular occlusions. Rodent models of I/R injury are providing significant insights into mechanisms and treatment strategies for human I/R injury, especially with regard to neurodegenerative damage in the retinal neurovascular unit. Presented here is a protocol for inducing retinal I/R injury in mice through elevation of intraocular pressure (IOP). In this protocol, the ocular anterior chamber is cannulated with a needle, through which flows the drip of an elevated saline reservoir. Using this drip to raise IOP above systolic arterial blood pressure, a practitioner temporarily halts inner retinal blood flow (ischemia). When circulation is reinstated (reperfusion) by removal of the cannula, severe cellular damage ensues, resulting ultimately in retinal neurodegeneration. Recent studies demonstrate inflammation, vascular permeability, and capillary degeneration as additional elements of this model. Compared to alternative retinal I/R methodologies, such as retinal arterial ligation, retinal I/R injury by elevated IOP offers advantages in its&#160;anatomical specificity, experimental tractability, and technical accessibility, presenting itself as a valuable tool for examining neuronal pathogenesis and therapy in the retinal neurovascular unit.

This article describes a procedure for inducing retinal ischemia-reperfusion injury by elevated intraocular pressure in mice. Retinal ischemia-reperfusion injury by elevated intraocular pressure serves to model human pathologies characterized by compromised oxygen and nutrient delivery in the retina, enabling researchers to examine potential cellular mechanisms and treatments for human diseases of the retinal neurovascular unit.

视网膜缺血再灌注损伤的小鼠模型通过眼压升高

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

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

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 reduced blood supply to the heart, resulting in the loss of oxygen to and the ensuing necrosis of the heart muscle. The MI model has gained popularity for its use as a short-term ischemia-reperfusion model and a long-term permanent ligation model. Below, we describe a reliable method for the permanent ligation of the LAD. With mouse genetic engineering technology becoming more advanced, and with an increasing availability of quality murine surgical instruments, the mouse has become a popular model for MI surgeries. Our surgical model incorporates the use of an easily reversible anesthetic for the rapid recovery of the mouse; a minimally invasive endotracheal intubation without involving a tracheotomy; and a thoracentesis through the original thoracotomy site without creating an additional incision in the chest, as is done in some other methods, to effectively remove excess blood and air from the chest cavity. This method is comparatively less invasive than other methods, which dramatically reduces surgical and post-surgical complications and mortality and improves reproducibility.

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

We provide a reliable method for left anterior descending artery (LAD) ligation in a mouse model. This method is comparatively less invasive than other methods, involving endotracheal intubation, a left-sided thoracotomy approach, and thoracentesis. This method can be used as a model for both acute and chronic myocardial infarction (MI).

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 ...

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 ...

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

Myocardial infarction (MI) and acute coronary diseases are among the most prominent causes of death in population with western lifestyle. The murine models of MI with permanent ligation of left-anterior descending (LAD) coronary artery closely mimics MI in humans. Murine models benefit from the extensive genetic engineering available nowadays. Here we propose a reproducible murine surgical model of myocardial infarction by permanent LAD coronary ligation. Our technique comprises anesthesia with ketamine/xylazine that can be rapidly reversed by administration of an antagonist, intubation without tracheotomy for mechanical-assisted ventilation, ventilation with application of extrinsic positive end-expiratory pressure (PEEP) to avoid alveolar collapse, a thoracotomy method limiting to the minimum surgical lesions made to skeletal muscles, and lung inflation without thoracentesis. This method is sparsely invasive, reproducible and reduces post-surgery mortality and complications.

Herein we describe a surgical procedure showing how to achieve permanent ligation of the left-anterior descending coronary artery in mice. This model is of high relevance to investigate the pathophysiology of myocardial infarction and the concomitant biological processes.

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 ...

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, ...

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

温缺血/再灌注损伤的大鼠肺移植模型：改善结局的优化

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...

Induction of Myocardial Infarction and Myocardial Ischemia-Reperfusion Injury in Mice

Acute myocardial infarction is a common cardiovascular disease with high mortality. Myocardial reperfusion injury can counteract the beneficial effects of heart reflow and induce secondary myocardial injury. A simple and reproducible model of myocardial infarction and myocardial ischemia-reperfusion injury is a good tool for researchers. Here, a customizable method to create a myocardial infarction (MI) model and MIRI by precision ligation of the left anterior descending coronary artery (LAD) through micromanipulation is described. Accurate and reproducible ligature positioning of the LAD helps obtain consistent results for heart injury. ST-segment changes can help to identify model accuracy. The serum level of cardiac troponin T (cTnT) is used to assess the myocardial injury, cardiac ultrasound is employed to evaluate the myocardial systolic function, and Evans-Blue/triphenyl tetrazolium chloride staining is used to measure infarct size. In general, this protocol reduces procedure duration, ensures controllable infarct size, and improves mouse survival.

Here we describe a simple and reproducible method that can induce myocardial infarction or myocardial ischemia-reperfusion injury in mice by precision ligation of the left anterior descending coronary artery through micromanipulation.

小鼠心肌梗死和心肌缺血再灌注损伤的诱导

Acute myocardial infarction is a common cardiovascular disease with high mortality. Myocardial reperfusion injury can counteract the beneficial effects of ...

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

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 order to explore the pathophysiological changes after myocardial infarction and design future treatments, research models of MI are required. Permanent ligation of the left coronary artery (LCA) in mice is a popular model to investigate cardiac function and ventricular remodeling post MI. Here we describe a less invasive, reliable, and reproducible surgical murine MI model by permanent ligation of the LCA. Our surgical model comprises of an easily reversible general anesthesia, endotracheal intubation that does not require a tracheotomy, and a thoracotomy. Electrocardiography and troponin measurement should be performed to ensure MI. Echocardiography at day 28 after MI will discern heart function and heart failure parameters. The degree of cardiac fibrosis can be evaluated by Masson's trichrome staining and cardiac MRI. This MI model is useful for studying the pathophysiological and immunological alterations after MI.

Presented here is a surgical procedure for permanent ligation of the left coronary artery in mice. This model can be used to investigate the pathophysiology and associated inflammatory response after myocardial infarction.

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 ...