我们感谢丹尼尔迪埃尔灌注固定技术为他的意见。

1。麻醉诱导<ol><li>异氟醚诱导，鼠标放到感应框，它是连接到蒸汽集到3.0％和0.5 L / min的氧流量。 </li><li>无意识与触觉的刺激，未能诱发反应和前肢或后肢踏板撤回反射缺席实现后，放置在一个温度控制的手术台鼠标在仰卧位。鼻锥通过感应框连接到蒸汽维持麻醉。放倒头固定用5-0尼龙线的上门牙，以方便插管。 </li><li>插入直肠温度探头的身体核心温度保持在37°C。用胶带固定四肢。越过较低的左腿在右腿开辟左胸部和更好地暴露心脏。 </li><li>脱毛霜适用于颈部和左前胸。 1分钟后擦去奶油。适用于局部皮肤碘伏消毒。皮下注入丁丙诺啡0.05毫克/千克体重，缓解疼痛。 </li><li>中线颈部皮肤切口用小剪刀。钝性解剖，包括气管腺体和肌肉。打开呼吸器。呼吸机的设置应调整生理参数。我们使用1 Minivent雨果萨克斯Elektronik公司，与哈佛仪器的呼吸频率105/min和潮气量200μL。一个镊子拉出舌头，轻轻地插入一个22政的金属管。确认气管内管和扩胸运动直视插管。 </li><li>绕过感应盒，开关，浪费天然气管道，以防止污染的实验室空间。蒸汽调整到2.0％。 </li></ol> 2。开胸手术<ol><li>请在左锁骨中线切开皮肤。钝性解剖对腋下皮下组织。确定的主要胸大肌肌肉的边界和钝剖析未成年人带够ectoralis肌肉下方。拉轻微的胸大肌肌肉的权利。您将获得的左肋直接观看。 </li><li>识别直言pentetrate与镊子第四肋间。让钳提示横跨肋间让自己插入是拉钩与橡皮筋附加到表的操作调整。你应该有一个明确的说法，包括心脏的心耳。这种访问通常没有任何失血，因而无电凝的需要。 </li></ol> 3。心的制备<ol><li>轻轻一拉，没有伤及心脏心包。 </li><li>提起从左心室前壁左心房耳廓识别左前降支（LAD）。法援署会被视为只在很短的模糊边界和鲜红的颜色相比静脉直当然。 </li></ol> 4。冠状动脉仪器仪表<ol><li>准备一个8-0的普理灵缝线形成U形，锥形针尖。通过足够深下法援署心肌针。 </li><li>切断缝合针在每边有1厘米的缝合结束。 </li><li>切断以防止任何尖角，封堵了1毫米PE-10管节。允许封堵器浸泡在酒精desinfection和挖掘使用前注意 ：管应浸泡24小时，100％的乙醇消毒适当4。 </li><li>主题既缝合结束通过封堵。 </li><li>使用大小3 Kalt缝合针引导两个缝合结束了上肋间。 </li></ol> 5。胸部闭幕<ol><li>连同6-0普理灵缝线配合打开肋间上下肋骨。关闭胸部之前，你应该继续到5.2。 </li><li> Hyperinflate肺部打开了几个呼吸周期夹紧呼气管起来肺不张。这个动作也可能取决于你使用的呼吸器类型。设置潮气量为300μL，直到胸部被关闭。 </li><li>设置呼吸机潮气量恢复到200μL。 </li><li>配合8-0 LAD结扎缝合两端，让他们形成了一个循环。 </li><li>附上一个心电图。 </li><li>挂钩的权重8-0循环，让小心挂的重量。你应该看到，在短短的心跳1显着的ST段抬高。释放的重量注 ：这些实验中使用了5.5重量g，但基于应变和小鼠的体重，体重可能会有所不同。 </li><li>放置在皮下口袋里的循环，并关闭6-0单结缝合皮肤。 </li><li>让鼠标拔管下变暖灯后恢复。 </li></ol> 6。心肌缺血再灌注<ol><li>经过了至少5天的恢复期诱导麻醉与氯胺酮，甲苯噻嗪的混合物和阿托品（4毫升/公斤体重，氯胺酮10毫克/毫升，甲苯噻嗪2毫克/毫升，阿托品0.06毫克/毫升，1。 </li><li>缺血再灌注损伤的实验与房间空气插管和通风。 </li><li>打开胸部缝合皮肤。准备8-0缝合循环。附上一个心电图。 </li><li>挂钩的权重，让他们挂。按照缺血协议。观看心电图ST段抬高2电位溶出。 </li><li>松开在缺血年底的重量。接近皮肤，拔管鼠标，让它恢复。 </li></ol> 7。再灌注时心肌梗死面积评估，最多3天<ol><li>麻醉及插管所需的再灌注时间结束的鼠标。 </li><li>在中线的xyphoid削减的胸部皮肤。打开腹部，左肋下切的隔膜。切断胸部锁骨中线两侧开放。 </li><li>修正获得畅通无阻的心脏缝合拍着前胸壁。 </li><li>仔细准备8-0缝合循环的。切循环和打结闭塞法援署。 </li><li> 10％的酞菁蓝注入左心房。为了防止心脏容量负荷，注入染料缓慢，不时吸。 </li><li>左心房注入氯化钾。平等梗死面积评估，这将逮捕的心脏在舒张。 </li><li>切出的心，留下尽可能多尽可能extracardial组织切割的心脏，以纾缓。 </li><li>在磷酸盐缓冲液洗的心脏。 </li><li>冻结在戊烷和液氮的心。此外，心可放入冰箱，直到轻轻冻结。 </li><li>切1毫米片的心脏。我们有刀片削减等于片（ 图4）心脏切片设备。确保正确对齐减少心脏的长轴垂直的心脏。 </li><li>在37°C孵育为1.5％，交委会在20分钟的片。我们使用96孔板每个切片放入一个孔。这将节省TTC和饶你使用的滤纸过滤器，以防止文物。 </li><li>用4％甲醛固定切片过夜。这将缩小片，但提高染料的对比度。 </li><li>切片放在显微镜幻灯片。盖上另一张幻灯片。使用在每个幻灯片年底1毫米的金属垫片和一起举行回形针幻灯片。 </li><li>从每个切片双方的数字图像。总是使用相同的设置和不小片放大。 </li><li>使用软件planimetry。我们使用由NIH ImageJ的。梗死区始终使用相同的标准，例如，只白色区域梗塞。白色粉红色区域梗塞。我们已经失明以及干预和planimetry的调查。 </li></ol> 8。组织学的替代心脏的制备<ol><li>按照步骤7.1至7.2。并继续到8.2。可于7可靠TTC染色梗死评估2小时再灌注，因为疤痕收缩。 </li><li>准备任何extracardial组织，钝性解剖覆盖主动脉根部的胸腺。 </li><li>抓住用钳子升主动脉和切出的心脏，为尽可能减少心外组织。 </li><li>洗净，轻轻挤在停搏液的心脏。 </li><li>放入停搏液填补了P35菜心。 </li><li>准备升主动脉。 </li><li> Cannulate甲醛预填充的一个套管升主动脉。我们使用锌福尔马林固定液24Ğ四线3条 。 </li><li>切孔之间的左心房和左心房。 </li><li>左心房连接到一个16厘米长的油管插入26Ğ导管。您还可以使用PE50管材。 </li><li>灌注福尔马林固定液10分钟的心。 </li><li>放入充满定影液最长为24小时7°C。管心</li><li>继续与准备组织学/免疫组织化学aration。 </li></ol> 9。代表结果慢性冠状动脉结扎术是一种复杂的技术，具有多个缺陷。然而，一旦掌握了它可以用非常低死亡率和高可靠的结果执行。小鼠和访问的心脏的最佳定位是成功识别和法援署仪器的关键。结扎的位置，显然会影响心肌梗死面积，因此需要有一个标准化的结扎部位。此外，如果间隔支的影响，这可能导致束支传导阻滞，ST段抬高。从epimyocardial静脉或从脑室出血，如果结扎太深，可能会出现和老鼠如果出血过多，应排除在外。 pericard应尽可能完整地被删除。离开pericard将加剧推入为结扎myocard的针。此外，它会引起心包炎，最终引起粘连，将很难做病理检查。 PE-封堵不应该有任何尖角，尽可能短，以尽量减少创伤，以myocard。防止胸部闭合后张力性气胸，肺恶性通货膨胀是绝对关键。有没有必要为胸腔引流。测试开胸结扎缝合两端拉一个正确的位置应该被忽略，因为拉张力是难以控制。如果法援署仪表失败，应避免进一步的尝试，因为这将增加心肌的创伤和水肿。 为了实现可靠的结果，应该是标准化的协议参数。因此，小鼠气管插管和通风与室内空气和体温紧紧控制反馈系统。已使用吊重已经强调。其他牵引装置有张力损失和非标准化的拉张力的缺点。缺血再灌注损伤和闭塞的多个周期的前和后处理的协议更容易进行吊重，因为他们只需要被解除，让挂（ 图1）。 梗死区（白色）应该区分风险（红色）和风险（蓝色）（ 图2A-B）的非区的地区。梗塞的大小取决于缺血时间。再灌注时间应该是至少2个小时，以便成功TTC染色（ 图1和2）。最重要的是，细胞因子的RNA表达低，在深水动物有动物进行心肌梗死（ 图3A-C的 ）相比，除缺血再灌注手术操作。 <img alt="图1" src="/files/ftp_upload/3896/3896fig1.jpg" /> 图1。梗塞面积的百分比（IS / AAR％）风险的大小。小鼠接受30分钟的缺血120分钟再灌注（I / R，N = 10）。的IPC：缺血后，小鼠接受缺血再灌注/闭塞20秒3个周期30分钟（N = 6，*表示P &lt;0.05）。 <img alt="图2A" src="/files/ftp_upload/3896/3896fig2A.jpg" /> 图2A。代表TTC -染色的心脏切片。白：心肌梗死面积，红色：危险区，蓝：非闭塞地区。 <img alt="图2B" src="/files/ftp_upload/3896/3896fig2B.jpg" /> 图2B。代表片的梗死面积（白色）。请注意，由于左心室接近顶点的圆锥形，epimyocard将出现平面面积，不应考虑平面测量测量（粉色/蓝色区外）。红色= TTC染色可行myocard。 <img alt="图3A" src="/files/ftp_upload/3896/3896fig3A.jpg" /> 图3A我无显着差异。缺血30分钟后，120分钟再灌注ocardial肿瘤坏死因子-αmRNA的表达。每组n = 4-6。 <img alt="图3B" src="/files/ftp_upload/3896/3896fig3B.jpg" /> 图3B。缺血30分钟后，120分钟再灌注（I / R）心肌IL-1β的mRNA表达。有没有控制之间的显著差异（不手术）和假手术（无脑缺血/再灌注组）。每组n = 4-6，*表示P &lt;0.05。 <img alt="图3C" src="/files/ftp_upload/3896/3896fig3C.jpg" /> 图心肌3C。的IL-6，30分钟后缺血mRNA表达和120分钟再灌注（I / R）。有没有控制之间的显著差异（不手术）和假手术（无脑缺血/再灌注组）。每组n = 4-6，*表示P &lt;0.05。 <img alt="图4" src="/files/ftp_upload/3896/3896fig4.jpg" /> 图4A-C。听取T 17111-1997切削设备。答：用刀片在切割位置封闭。 B：开放，侧视图。 ：开放，前视图。心将在其长轴垂直的刀片（箭头）的白色塑料床的凹槽对齐。

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没有利益冲突的声明。

我们已经修改通过访问通过左侧肋间开胸的心脏和领导法援署缝线，在左锁骨中线胸部封闭胸小鼠模型。保持骨左肋不变，将最大限度地减少创伤，需要止痛药物，外科手术部位感染，从而，促进恢复。通过保留左动脉内部哺乳动物有没有电灼需要。我们离开以后方便缝合皮下组织循环及使用吊重系统定义的闭塞。胸外模型使心肌缺血再灌注的所有研究方案的应用与手术创伤和随后的免疫反应<sup...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>汽</td><td> drägerwerk公司</td><td> isoflo </td></tr><tr><td>显微镜</td><td>徕卡</td><td> M80的</td></tr><tr><td>光源</td><td>肖特</td><td>吉隆坡1500液晶</td></tr><tr><td> homeothermic毯控制单元</td><td>哈佛仪器</td><td></td></tr><tr><td> MiniVent类型845 </td><td> HUGO高盛Elektronik公司</td><td></td></tr><tr><td> 8-0普理灵</td><td>爱惜康</td><td> BV130-56.5毫米3/8c </td></tr><tr><td> 6-0普理灵</td><td>爱惜康</td><td> BV-19.3毫米3/8c </td></tr><tr><td> kalt缝合针大小为3 </td><td>福斯特</td><td> 12050-03 </td></tr><tr><td>三苯</td><td> Sigma Aldrich公司</td><td> 93145 </td&gt; </tr><tr><td>酞菁蓝</td><td> Heubach GmbH德国有限公司</td><td></td></tr><tr><td> PowerLab系统</td><td> ADInstruments </td><td></td></tr></tbody></table>

a murine closed-chest model of myocardial ischemia and reperfusion

开胸冠状动脉结扎模型开胸手术创伤修改不同的机制参与缺血再灌注诱导的免疫反应。免疫反应包括细胞因子的表达和释放或分泌的内源性配体的先天免疫受体。先天免疫系统的激活可能调节心肌梗死面积。我们已经修改现有的小鼠封闭胸部模型，使用吊重研究心肌预处理和后处理和先天免疫在心肌缺血再灌注损伤中的作用，这可能是有用的。这种模式使动物心肌缺血的发作前从手术创伤恢复。 挥发性麻醉药已激烈的研究和预处理缺血心脏的影响是众所周知的。然而，这种保护作用排除在开胸冠状动脉结扎模型，它的使用。因此，另一个优势是次使用Ë以及可控的挥发性麻醉药仪器在慢性闭胸模型，因为他们的预处理效果持续长达72小时。慢性心脏疾病，间歇性缺血，多打款是这种模式的其他可能的应用。 为慢性闭胸模型，气管插管和通风的小鼠接受外侧钝通过的第4肋间开胸。继左前降连字前的鉴定，通过下面的船只都缝合两端螺纹通过封堵。然后，两个缝合两端都通过胸壁，打结，形成一个循环，并留在皮下组织。胸部关闭和恢复为5天之后，老鼠再次麻醉，胸部皮肤被重新吊重迷上了心电图控制下的循环。 在缺血/再灌注协议结束时，心中可以与TTC染色的梗塞大小评估或进行灌注固定，允许​​除了组织学和免疫组化形态的研究。

Watch this Scientific Journal Video about A Murine Closed-chest Model of Myocardial Ischemia and Reperfusion at JoVE.com

A Murine Closed-chest Model of Myocardial Ischemia and Reperfusion

手术创伤引起的炎症反应。被称为细胞因子和内源性配体调节缺血再灌注后心肌梗死面积。我们提出了小鼠缺血及使用吊重的再灌注，以减少开胸手术的效果修改封闭胸部模型。

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 ...

a-murine-closed-chest-model-of-myocardial-ischemia-and-reperfusion

Research

JoVE Journal

Medicine

28.6K 次观看.  University of Bonn, Germany. 手术创伤引起的炎症反应。被称为细胞因子和内源性配体调节缺血再灌注后心肌梗死面积。我们提出了小鼠缺血及使用吊重的再灌注，以减少开胸手术的效果修改封闭胸部模型。

视频: A Murine Closed-chest Model of Myocardial Ischemia and Reperfusion

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

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Background 
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered advancements to decrease the occurrence this injury.&#160;The current model demonstrates reproducible lower&#160;extremity paralysis following thoracic aortic occlusion.
Methods 
Adult male C57BL6 mice were anesthetized with isoflurane. Through a cervicosternal incision the aorta was exposed. The descending thoracic aorta and left subclavian arteries were identified without entrance into pleural space. Skeletonization of these arteries was followed by immediate closure (Sham) or occlusion for 4 min (moderate ischemia) or 8 min (prolonged ischemia). The sternotomy and skin were closed and the mouse was transferred to warming bed for recovery.&#160; Following recovery, functional analysis was obtained at 12 hr intervals until 48 hr.
Results 
Mice that underwent sham surgery showed no observable hind&#160;limb deficit. Mice subjected to moderate ischemia for 4 min had minimal functional deficit at 12 hr followed by progression to complete paralysis at 48 hr. Mice subjected to prolonged ischemia had an immediate paralysis with no observable hind-limb movement at any point in the postoperative period. There was no observed intraoperative or post&#160;operative mortality.
Conclusion 
Reproducible lower extremity paralysis whether immediate or delayed can be achieved in a murine model. Additionally, by using a median sternotomy and careful dissection, high survival rates, and reproducibility can be achieved.

The lack of mechanistic understanding of spinal cord ischemia-reperfusion injury has hindered further adjuncts to prevent paraplegia following high risk aortic operations. Thus, the development of animal models is imperative. This manuscript demonstrates reproducible lower extremity paralysis following thoracic aortic occlusion in a murine model.

可重放截瘫胸主动脉阻断脊髓缺血再灌注的小鼠模型

Background
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered ...

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 ...

A Rat Model of Ventricular Fibrillation and Resuscitation by Conventional Closed-chest Technique

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic components of cardiopulmonary resuscitation in humans is herein described. The model was developed in 1988 and has been used in approximately 70 peer-reviewed publications examining a myriad of resuscitation aspects including its physiology and pathophysiology, determinants of resuscitability, pharmacologic interventions, and even the effects of cell therapies. The model featured in this presentation includes: (1) vascular catheterization to measure aortic and right atrial pressures, to measure cardiac output by thermodilution, and to electrically induce ventricular fibrillation; and (2) tracheal intubation for positive pressure ventilation with oxygen enriched gas and assessment of the end-tidal CO2. A typical sequence of intervention entails: (1) electrical induction of ventricular fibrillation, (2) chest compression using a mechanical piston device concomitantly with positive pressure ventilation delivering oxygen-enriched gas, (3) electrical shocks to terminate ventricular fibrillation and reestablish cardiac activity, (4) assessment of post-resuscitation hemodynamic and metabolic function, and (5) assessment of survival and recovery of organ function. A robust inventory of measurements is available that includes &#8211; but is not limited to &#8211; hemodynamic, metabolic, and tissue measurements. The model has been highly effective in developing new resuscitation concepts and examining novel therapeutic interventions before their testing in larger and translationally more relevant animal models of cardiac arrest and resuscitation.

This article describes a rat model of electrically-induced ventricular fibrillation and resuscitation by chest compression, ventilation, and delivery of electrical shocks that simulates an episode of sudden cardiac arrest and conventional cardiopulmonary resuscitation. The model enables gathering insights on the pathophysiology of cardiac arrest and exploration of new resuscitation strategies.

室颤由传统的封闭胸技术大鼠模型和复苏

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic ...

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, ...

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.

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 ...

Improvement of a Closed Chest Porcine Myocardial Infarction Model by Standardization of Tissue and Blood Sampling Procedures

Myocardial ischemia reperfusion (I/R) injury contributes to almost half of the necrotic area after myocardial infarction. To date there is no approved drug to prevent or reduce myocardial I/R injury. The study and understanding of the pathophysiological mechanisms of myocardial I/R injury is essential to develop successful treatments. Large animal experiments are an important step in translational methods. The porcine model of acute myocardial infarction has been established and described by ourselves and others. We aimed to further improve the value of the model by focusing in detail on the sampling techniques for use in future experiments. Furthermore, we emphasize small but important steps that can affect the quality of the final results. To mimic the clinical situation of myocardial I/R injury, a percutaneous coronary intervention (PCI) catheter was inserted into the left anterior descending coronary artery (LAD) of an anesthetized pig. &#176;&#176;&#176;This model mimics acute myocardial infarction and PCI treatment in humans with the possibility of accurately determining the area at risk as well as the necrotic- and viable ischemic tissue. Here the model was used to investigate the effect of a bicyclic peptide inhibitor of FXIIa. The model can also be modified to allow longer reperfusion times to study later effects of myocardial infarction.

Here we demonstrate a protocol to standardize sampling procedures of an established porcine model of acute myocardial infarction in order to increase its translational value in the understanding of the pathophysiology of myocardial ischemia/reperfusion injury and to test novel drug candidates.

组织和采血程序标准化改善闭合性胸猪心肌梗死模型的研究

Myocardial ischemia reperfusion (I/R) injury contributes to almost half of the necrotic area after myocardial infarction. To date there is no approved ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to perform a partial thoracotomy to visualize the transverse aortic arch. However, the surgical trauma caused by the thoracotomy in open-chest models changes the respiratory physiology as the ribs are dissected and left unattached after chest closure. To prevent this, we established a minimally invasive, closed chest approach via lateral thoracotomy. Herein we approach the aortic arch via the 2nd intercostal space without entering the chest cavities, leaving the mouse with a less traumatic injury to recover from. We perform this operation using standard laboratory settings for open chest TAC procedures with equal survival rates. Apart from maintaining physiological breathing patterns due to the closed chest approach, the mice seem to benefit by showing rapid recovery, as the less invasive technique appears to facilitate a fast healing process and to reduce immune response after trauma.

Here, we present a protocol of Transverse Aortic constriction (TAC) via a lateral thoracotomy. This technique is a minimally invasive, closed chest surgical procedure aiming to simulate pressure overload and heart failure in mice utilizing standard TAC laboratory settings.

诱导小鼠横向主动脉狭窄的闭合胸模型

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...

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 ...

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, ...