是由俄罗斯科学基金会支持这项工作，不授予。 14-50-00060和LLC RUSCHEMBIO。这项工作也是由美国Crohn`s和结肠炎基金会授予294083（至AVT），由美国国立卫生研究院授予RO1 DK47700（CJ到）的支持。

动物研究按照健康准则全国学院进行，并由机构动物护理被批准，并使用特鲁多研究所委员会。 8-12周龄的C57BL / 6小鼠用于该研究。 1.准备手术<ol><li>准备和消毒手术器械。 </li><li>制备基于异氟醚麻醉系统的鼻锥，并加热垫。确保加热垫不会过热（&lt;39℃）。 </li><li>确保异氟烷气体清除罐被正确地定位，以确保排气口在所述罐的底部不被堵塞或以任何方式闭塞。称重之前对罐程序和文件重气体清扫罐。当罐的重量超过50g累计重量（〜12小时）的丢弃罐。 </li></ol> 2.麻醉<ol><li>麻醉小鼠，用3％异氟烷在感应腔室（1升/分钟O 2）。 <ol><li>通过无力评估麻醉深度，以保持直立，有目的的自主运动，眨眼反射消失，肌肉松弛，以及失去响应的损失反射性刺激（趾或尾掐以坚定的压力）。 </li><li>通过观察胸壁和腹部的运动评估呼吸频率和模式。在最佳麻醉，呼吸速率应为每分钟55-65〜呼吸。 </li><li>从感应腔除去小鼠，并迅速剃鼠标的腹部区域。 </li></ol></li><li>为预防角膜干燥，将平淡无奇的眼药膏在眼里。 </li><li>放置在加热垫鼠标，它通过鼻锥连接到麻醉系统。确保胶乳鼻锥膜牢固地配合在小鼠头部并没有异氟烷的泄漏。 </li><li>减少异氟烷率至1.5％，并注入丁丙诺啡（0.1毫克/千克）和氯胺酮（10毫克/千克）经皮下以防止疼痛级联的卷绕。 </li><li>擦拭日e皮肤与手术优碘溶液，然后用70％乙醇浸泡消毒棉签操作区。 </li></ol> 3.手术<ol><li>做一个中线3-5厘米剖腹手术以经营剪刀。用无菌非粘附垫用生理盐水沾湿覆盖操作区域。隔离盲肠和回肠，用在盐水中浸湿的棉签暴露肠系膜上动脉。 </li><li>为了方便剪辑应用，在周围用细光圈scissors.To做到这一点，肠系膜上动脉肠系膜做小缺口，轻轻抬起换药镊子肠道，并在需要的剪辑的位置上切断肠系膜上动脉两侧肠系膜（ 图1A）。然后，施加片段之前添加无菌生理盐水几滴需要的剪辑的位置的区域。 注意:要进行假手术，按照手术步骤执行到步骤3.2。不适用剪辑。相反，保持组织通过添加温暖湿润小号艾琳在3.6 1小时描述。随后，执行步骤4.1， </li><li>闭塞使用一个夹具施放创建相邻盲肠（ 图1B）的缺血性回肠的5-7厘米区域微血管剪辑（70挤压力g）肠系膜上动脉的第一级分支。虽然船只的位置是保守的，有可能是小鼠之间的细微变化（参见图1的例子）。因此，2或3个片段，通常需要（参见图1A，D，E，黑色箭头的剪辑的位置）。 注意:使用高品质的容器剪辑。高压剪辑会损害血管和防止再生，而低压剪辑（&lt;30克）可以不完全阻断血流。 </li><li>通过使用跨血管2微血管夹子（40克的力）的肠阻塞侧支血流，demarking缺血肠的区域（ 图1）。侧支血管闭塞，需要防止从相邻血管的血液供应（参见图1A，D，E，绿色箭头的剪辑的位置）。 </li><li>可选:添加肝素溶液（6 USP单位/ ml），以防止血液凝固。滴加到0.5肝素溶液毫升的离体肠管。 </li><li>用盐水湿无菌非粘附垫娇嫩湿巾预先加热到37℃，并适用于手术区。确保湿巾在整个过程中保持湿润。 </li><li>保持缺血使用整个1-1.5％异氟烷麻醉60分钟。如果正确地执行局部缺血过程中，局部缺血区域将在大约30分钟改变为酒红色的颜色。需要注意的是血管远侧夹子位置指示成功闭塞缺血（ 图1，右图）中被放大。 </li><li>密切监察在缺血阶段鼠标。继续适用盐溶液到非粘附垫覆盖所述手术部位。 </li><li>标记缺血区的通过在组织上吸取20微升Gill`s 3苏木的边缘，以方便收获从同一小鼠的缺血组织和邻近的健康组织进行比较（ 图1E，右图）。 </li></ol> 4.再灌注阶段<ol><li>在缺血的末尾添加上剪辑区域生理盐水滴几滴，轻轻取出微血管剪辑施夹。然后，用盐水湿润棉提示轻轻地推肠道回腹腔。除去非贴壁垫和关闭使用9个毫米的不锈钢伤口夹腹壁和皮肤。如果超过3小时进行再灌注时间越长，使用可吸收的薇乔缝合皮肤上施加缠绕剪辑之前关闭腹壁。 </li><li>保持在加热的清洁笼子小鼠的灌注阶段的时间所需量的（30分钟，60分钟，120分钟，或180分钟）。 </li><li>检查动物至少每30分钟，以确保稳定性。 </li></ol><p类="jove_title"&gt; 5。剖检及小肠的收获<ol><li>用CO安乐死的小鼠2过量随后在所希望的时间再灌注颈脱位。 </li><li>打开腹腔并收集作进一步的分析缺血性肠组织。邻接受伤组织作为内部对照以考虑到损伤任何全身反应收获健康的正常组织。 注意:此控件是比较合适的比假手术的对照组小鼠，因为假手术小鼠不接受以IR-性损伤全身反应。 </li><li>洗出用30毫升注射器填充以生理盐水，然后连接灌胃针切肠肠纵向内容。如果需要对基因表达分析小肠的样品，切1.5毫米片段纵向，并用于组织学分析的剩余片。 </li><li>对于组织学分析，使用一对镊子来滚动INTEST准备了瑞士卷国家统计局。 </li><li>维持轧制形式，放置肠中的多条组织盒活检泡沫垫（ 图2）之间。放置在10％缓冲福尔马林卡匣。 </li><li>修复组织在福尔马林中至少24小时。额外24小时替换用70％乙醇福尔马林。商店组织在70％乙醇无限期在室温下。 </li><li>在石蜡中嵌入，切为5μm切片并用苏木精和曙红使用标准协议（图3）染色。 </li></ol> 6.评分<ol><li>分数鼠缺血再灌注损伤如表2。选择一个合适的评分方法。 </li><li>可选:划分的视场分为四个部分，因为伤害的严重程度在整个一节有所不同。 </li><li>从计算得到盲目得分每一部分的平均品位。 </li><li>比较案件和控制之间的受损组织的档次以及为thE使用秩和检验测试，随后邓恩的多重比较试验没有受伤的组织。 </li></ol>

The authors declare no conflict of interest

肠道缺血再灌注损伤的小鼠模型的发展大大提高了组织损伤机制的理解和潜在的治疗策略的发展援助，以尽量减少组织损伤7,9,11,34。此协议的关键步骤是微血管剪辑的正确定位，缺血再灌注损伤的缺血和适当的组织学评价的正确时机。 缺血的持续时间是为后续上皮损伤是至关重要的。以诱导可重复的IR损伤无发病率和实验小鼠的死亡率通常所需的时间是45-60分钟，随?...

肠道是引起缺血和上皮损伤的血流量的中断非常敏感。缺血再灌注提供了组织的复氧，并能进一步促进病理。因此，肠缺血再灌注损伤是具有广泛的病状，包括坏死性小肠结肠炎，在小肠移植同种异体移植物排斥，腹主动脉瘤手术，心肺旁路和炎性肠病1,2-并发症相关联。肠缺血再灌注损伤，尤其是急性肠系膜缺血，是造成发病率和死亡率3危及生命的疾病。 虽然了解甚少，肠缺血再灌注（IR）的损伤被认为是与肠道菌群的变化以及产生活性氧和炎性细胞因子和趋化因子1,4-6相关联。这导致在这两种激活内特和促进炎症和组织损伤1,7,8适应性免疫的机制。 动物模型对于理解IR损伤的机制的关键，因为它们允许容易GAIN-和失功能遗传实验。已开发红外几种动物模型，其中包括完整的血管闭塞，低流量缺血和分段血管闭塞（在最近的一次全面回顾总结9）。引起肠系膜上动脉（SMA）的完整的血管闭塞肠缺血是IR的大型动物和啮齿动物9-11一个简单而常用的模式。然而，肠道的不同区域有不同的易感性伤害。此外，各种不同的麻醉剂，镇痛药，动脉阻塞技术，以及在可变程度的损伤缺血性损伤和恢复结果的持续时间不一致混杂我们的红外跨多个studie的生物学的理解秒。 表1展示了在小鼠IR研究这些不一致。从使用较短的缺血时间（30-45分钟），最大的缺点是针对在可观察的病例与对照组之间差异明显复苏的窗口。轻度损伤的上皮可以解决再灌注后一小时，因此专门病理指标可能需要找到在上皮恢复原状的差异。与此相反，过大的损坏，由100分钟缺血损伤所看到可能导致上皮，恢复原状不再可能的完整denudement，增加死亡率，和恢复时间。因此，在这里我们描述肠道IR小鼠的详细程序导致重复性的伤害死亡率不鼓励在我们领域这一技术的标准化。肠IR损伤的该模型可用于研究损伤和再生的细胞和分子机制。

<table><tbody><tr><td>Heated Pad</td><td>Sunbeam</td><td>E12107-819</td><td>Alternative: Braintree Scientific heated pad</td></tr><tr><td>Table top research anesthesia Machine</td><td>Vasco</td><td>UCAP 0001-0000171</td><td>Alternative: Parkland Scientific, V3000PS</td></tr><tr><td>Nose Cone</td><td>Parkland Scientific</td><td>ARES500</td><td></td></tr><tr><td>Scavenger canister and replacement cartridge</td><td>Parkland Scientific</td><td>80000, 80120</td><td></td></tr><tr><td>Induction Chamber</td><td>Surgivet</td><td>V711802</td><td></td></tr><tr><td>Isoflurane</td><td>Piramal Healthcare</td><td>NDC 66794-013-10</td><td>Controlled substance, contact IACUC</td></tr><tr><td>Animal clipper&amp;nbsp;</td><td>Oster&amp;nbsp;</td><td>Oster Golden A5 078005-050-003</td><td></td></tr><tr><td>Ophthalmic ointment</td><td>Webster</td><td>8804604</td><td></td></tr><tr><td>Buprenorphine</td><td>McKesson</td><td>562766</td><td>Controlled substance,contact IACUC</td></tr><tr><td>Ketaset (Ketamine HCl)</td><td>Pfizer</td><td>NADA 45-290</td><td>Controlled substance, contact IACUC</td></tr><tr><td>Cotton tips</td><td>Puritan medical products</td><td>806-WC</td><td>Autoclave before use</td></tr><tr><td>Betadine</td><td>Purdue Products</td><td>67618-150-17</td><td>10% Povidone-Iodine</td></tr><tr><td>Sterile saline solution</td><td>Aspen</td><td>46066-807-60</td><td>Adjust to room temperature before use</td></tr><tr><td>IR rodent thermometer</td><td>BIOSEB</td><td>BIO-IRB153</td><td></td></tr><tr><td>Micro vascular clips, 70 g</td><td>Roboz Surgical&amp;nbsp;</td><td>RS5424, RS5435</td><td>Alternative: WPI 14121, for SMA occlusion</td></tr><tr><td>Micro vascular clips, 40 g</td><td>Roboz Surgical&amp;nbsp;</td><td>RS6472</td><td>Alternative:WPI 14120, for collateral vessels occlusion</td></tr><tr><td>Clip applying forceps</td><td>World Precision Instruments</td><td>14189</td><td>Alternative: Roboz #RS-5410 or&amp;nbsp; #RS-5440</td></tr><tr><td>Gill&#039;s 3 hematoxylin</td><td>Thermo Scientific</td><td>14-390-17</td><td></td></tr><tr><td>Surgical staples, Reflex 9 mm</td><td>Cell Point Scientific</td><td>201-1000</td><td></td></tr><tr><td>Autoclip applier</td><td>Beckton Dickinson</td><td>427630</td><td></td></tr><tr><td>Byopsy foam pad</td><td>Simport</td><td>M476-1</td><td></td></tr><tr><td>Tissue cassette</td><td>Fisher Healthcare</td><td>15182701A</td><td>Histosette II combination lid and base</td></tr><tr><td>10% buffered formalin</td><td>Fisher Scientific</td><td>245-684</td><td></td></tr><tr><td>Surgical iris scissors</td><td>World Precision Instruments</td><td>501263-G SC</td><td>Alternative: Roboz RS6816</td></tr><tr><td>Operating scissors</td><td>World Precision Instruments</td><td>501219-G</td><td>Alternative: Roboz RS6814</td></tr><tr><td>Dressing forceps</td><td>Roboz Surgical&amp;nbsp;</td><td>RS-5228, RS-8122</td><td>Alternative: World Precision Instruments 1519-G</td></tr><tr><td>Heparin, endotoxin free, 300 USP units/vial, 50 mg</td><td>Sigma</td><td>2106</td><td></td></tr><tr><td>Reflex wound clip removing forceps</td><td>Roboz Surgical&amp;nbsp;</td><td>RS-9263</td><td>Alternative: World Precision Instruments: 500347</td></tr><tr><td>Mice C57BL/6J mice&amp;nbsp;</td><td>Jackson Laboratory</td><td>Stock No 0664</td><td></td></tr><tr><td>Telfa non-adherent dressings, 3 x 4, sterile</td><td>Coviden</td><td>1050</td><td></td></tr><tr><td>Fisherbrand transfer pipets</td><td>Fischer Scientific</td><td>13-711-5AM</td><td>Use pipets to dropwise add saline</td></tr></tbody></table>

murine model of intestinal ischemia-reperfusion injury

肠缺血是具有广泛的临床病症，包括动脉粥样硬化，血栓形成，低血压，坏死性小肠结肠炎，肠移植，创伤和慢性炎症有关的威胁生命的情况。肠缺血再灌注（IR）的损伤为急性肠系膜缺血的结果，通过肠系膜血管引起的血流量不足，导致肠损伤。再灌注缺血可能进一步加剧肠道损伤。 IR损伤的机制是复杂的和理解甚少。因此，实验小动物模型是了解缺血再灌注损伤的病理生理和新疗法的发展至关重要。 这里，我们描述急性肠道IR损伤，提供了小肠的重现性损伤而不死亡率的小鼠模型。这是通过诱导缺血远端回肠的区域由在时间上的occludin实现克用微血管夹子60分钟肠系膜上动脉的周边和终端侧枝。再灌注1小时后，或在通过组织学分析研究了肠的可再现伤伤结果后2小时。微血管剪辑的正确位置的程序是至关重要的。因此，视频剪辑提供这一技术的详细目视一步一步的说明。肠IR损伤的该模型可用于研究损伤和再生的细胞和分子机制。

我们优化的IR手术的实验方案获得小鼠回肠的重现性IR损伤的作用。代表性的结果证明了在此部分。 图1示出微血管剪辑位置的例子以诱导回肠的缺血。黑色箭头显示夹闭肠系膜上动脉一级分支的主要片段的位置。绿色箭头表示的附加片段的位置，以阻止来自侧支血管的血液供应。注意闭塞血管远端片段位置和intest...

Watch this Scientific Journal Video about Murine Model of Intestinal Ischemia-reperfusion Injury at JoVE.com

Murine Model of Intestinal Ischemia-reperfusion Injury

肠缺血再灌注损伤的小鼠模型

这里，我们描述肠缺血再灌注的小鼠中的详细过程，这导致在重现的损伤而不死亡率鼓励跨字段这种技术的标准化。肠缺血再灌注损伤的该模型可用于研究损伤和再生的细胞和分子机制。

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

murine-model-of-intestinal-ischemia-reperfusion-injury

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

19.4K 次观看.  Trudeau Institute. 这里，我们描述肠缺血再灌注的小鼠中的详细过程，这导致在重现的损伤而不死亡率鼓励跨字段这种技术的标准化。肠缺血再灌注损伤的该模型可用于研究损伤和再生的细胞和分子机制。 

视频: Murine Model of Intestinal Ischemia-reperfusion Injury

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 ...

科研

医学

Murine Model of Metastatic Liver Tumors in the Setting of Ischemia Reperfusion Injury

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce multiple tissue and organ damage. Despite recent advances and therapeutic approaches, the overall morbidity has remained unsatisfactory especially in patients with underlying parenchymal abnormalities. In the context of aggressive cancer growth and metastasis, surgical I/R is suspected to be the promoter regulating tumor recurrence. This article aims to describe a clinically relevant murine model of liver I/R and colorectal liver metastasis. In doing so, we aim to assist other investigators in establishing and perfecting this model for their routine research practice to better understand the effects of liver I/R on promoting liver metastases.

We describe in detail a clinically relevant colorectal cancer liver metastases (CRLM) tumor model and the influence of liver ischemia reperfusion (I/R) in tumor growth and metastasis. This model can help to better understand the mechanisms underlying surgery-induced promotion of liver metastatic growth.

缺血再灌注损伤中转移性肝肿瘤的模样

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce ...

癌症研究

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 ...

Intestinal Stem Cell Isolation and Culture in a Porcine Model of Segmental Small Intestinal Ischemia

Intestinal ischemia remains a major cause of morbidity and mortality in human and veterinary patients. Many disease processes result in intestinal ischemia, when the blood supply and therefore oxygen is decreased to the intestine. This leads to intestinal barrier loss and damage to the underlying tissue. Intestinal stem cells reside at the base of the crypts of Lieberk&#252;hn and are responsible for intestinal renewal during homeostasis and following injury. Ex vivo cell culture techniques have allowed for the successful study of epithelial stem cell interactions by establishing culture conditions that support the growth of three-dimensional epithelial organ-like systems (termed "enteroids" and "colonoids" from the small and large intestine, respectively). These enteroids are composed of crypt and villus-like domains and mature to contain all of the cell types found within the epithelium. Historically, murine models have been utilized to study intestinal injury. However, a porcine model offers several advantages including similarity of size as well as gastrointestinal anatomy and physiology to that of humans. By utilizing a porcine model, we establish a protocol in which segmental loops of intestinal ischemia can be created within a single animal, enabling the study of differing time points of ischemic injury and repair in vivo. Additionally, we describe a method to isolate and culture the intestinal stem cells from the ischemic loops of intestine, allowing for the continued study of epithelial repair, modulated by stem cells, ex vivo.

This protocol will enable readers to successfully establish a porcine model of segmental intestinal ischemia and subsequently isolate and culture intestinal stem cells for the study of epithelial repair following injury.

猪节段性小肠缺血模型中肠道干细胞的分离与培养

Intestinal ischemia remains a major cause of morbidity and mortality in human and veterinary patients. Many disease processes result in intestinal ...

Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse model is useful for investigating the pathophysiology of cerebral ischemia-reperfusion and determining potential therapeutic strategies for patients with stroke. Variations in the anatomy of the circle of Willis of C57BL/6 mice affects their infarct volume after cerebral-ischemia-induced injury. Studies have indicated that distal MCA occlusion (MCAO) can overcome this problem and result in a stable infarct size. In this study, we establish a two-vessel occlusion mouse model of cerebral ischemia-reperfusion through the interruption of the blood flow to the right MCA. We distally ligate the right MCA and right common carotid artery (CCA) and restore blood flow after a certain period of ischemia. This ischemia-reperfusion injury induces an infarct of stable size and a behavioral deficit. Peripheral immune cells infiltrate the ischemic brain within the 24 h infiltration period. Additionally, the neuronal loss in the cortical area is less for a longer reperfusion duration. Therefore, this two-vessel occlusion model is suitable for investigating the immune response and neuronal recovery during the reperfusion period after cerebral ischemia.

A mouse model of cerebral ischemia-reperfusion is established to investigate the pathophysiology of stroke. We distally ligate the right middle cerebral artery and right common carotid artery and restore blood flow after 10 or 40 min of ischemia.

脑缺血再灌注的双血管闭塞小鼠模型

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse ...

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 ...

An Effective Mouse Model of Unilateral Renal Ischemia-Reperfusion Injury

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models are the only available resources that mimic the complexities of the IRI-associated damage encountered in vivo. This paper describes an effective mouse model of unilateral renal IRI that delivers highly reproducible data. Ischemia is induced by occluding the right renal pedicle for 30 min followed by reperfusion. In addition to the surgical procedure, a sequential overview of the expected physiological and histopathological changes following renal IRI will be provided by comparing data from seven different reperfusion times (4 h, 8 h, 16 h, 1 day, 2 days, 4 days, and 7 days). Critical data for planning experiments ahead, such as mean surgical time, average anesthetic consumption, and body weight changes over time, will be shared. This work will help researchers implement a reliable renal IRI model and select the appropriate reperfusion time that aligns with their intended investigative goals.

Renal ischemia-reperfusion injury is associated with high morbidity and mortality in hospitalized patients. Here, we present a simple and effective mouse model of unilateral renal ischemia-reperfusion injury and provide a sequential overview of representative pathological changes observed in the kidney.

单侧肾缺血再灌注损伤的有效小鼠模型

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models ...

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 ...

Rodent Model of Intestinal Ischemia-Reperfusion Injury via Occlusion of the Superior Mesenteric Artery

Intestinal ischemia-reperfusion injury (IRI) is associated with a myriad of conditions in both veterinary and human medicine. Intestinal IRI conditions, such as gastric dilatation volvulus (GDV), mesenteric torsion, and colic, are observed in animals such as dogs and horses. An initial interruption of blood flow causes tissues to become ischemic. Although necessary to salvage viable tissue, subsequent reperfusion can induce further injury. The main mechanism responsible for IRI is free radical formation upon reperfusion and reintroduction of oxygen into damaged tissue, but there are many other components involved. The resulting local and systemic effects often impart a poor prognosis. 
Intestinal IRI has been the subject of extensive research over the past 50 years. An in vivo rodent model in which the base of the superior mesenteric artery (SMA) is temporarily ligated is currently the most common method used to study intestinal IRI. Here, we describe a model of intestinal IRI utilizing isoflurane anesthesia in 21% O2 medical air that yields reproducible injury, as demonstrated by consistent histopathology of the small intestines. Tissue injury was also assessed in the colon, liver, and kidneys.

We describe how to generate a widely used surgical model of intestinal ischemia-reperfusion injury (IRI) in rodents. The procedure involves occlusion of the superior mesenteric artery followed by the restoration of blood flow. This model is useful for studies investigating occlusive causes of intestinal IRI in both veterinary and human medicine.

肠系膜上动脉闭塞的肠缺血再灌注损伤的啮齿动物模型

Intestinal ischemia-reperfusion injury (IRI) is associated with a myriad of conditions in both veterinary and human medicine. Intestinal IRI conditions, ...

研究肺缺血再灌注损伤的小鼠模型

Murine Left Pulmonary Hilar Clamp Model of Lung Ischemia Reperfusion Injury

Ischemia reperfusion injury (IRI) during lung transplantation is a major risk factor for post-transplant complications, including primary graft dysfunction, acute and chronic rejection, and mortality. Efforts to study the underpinnings of IRI led to the development of a reliable and reproducible mouse model of left lung hilar clamping. This model involves a surgical procedure performed in an anesthetized and intubated mouse. A left thoracotomy is performed, followed by careful lung mobilization and dissection of the left pulmonary hilum. The hilar clamp involves reversible suture ligation of the pulmonary hilum with a slipknot, which stops the arterial inflow, venous outflow, and airflow through the left mainstem bronchus. Reperfusion is initiated by careful removal of the suture. Our laboratory uses 30 min of ischemia and 1 h of reperfusion for the experimental model in the current investigations. However, these time periods can be modified depending on the specific experimental question. Immediately prior to sacrifice, arterial blood gas can be obtained from the left ventricle after a 4 min period of right hilar clamping to ensure that the PaO2 values obtained are attributed to the injured left lung alone. We also describe a method to measure cell extravasation with flow cytometry, which involves intravenous injection of a fluorochrome-labeled antibody specific for the cell(s) to be studied prior to sacrifice. The left lung can then be harvested for flow cytometry, frozen or fixed, paraffin-embedded immunohistochemistry, and quantitative polymerase chain reaction. This hilar clamp technique allows for&#160;detailed study of the cellular and molecular mechanisms underlying IRI. Representative results reveal decreased left lung oxygenation and histologic evidence of lung injury following hilar clamping. This technique can be readily learned and reproduced by personnel with and without microsurgical experience, leading to reliable and consistent results and serving as a widely adoptable model for studying lung IRI.

作者聚焦:缺血再灌注对肺移植影响的见解

The protocol outlines a feasible, reliable, and reproducible method of left pulmonary hilar clamping that can be used to study lung ischemia-reperfusion injury in mouse models.