Name: renal ischaemia reperfusion injury: a mouse model of injury and regeneration
Uploaded: 2014-06-07T14:00:00.000+00:00
Duration: 12 min 27 s
Description: Watch this Scientific Journal Video about Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration at JoVE.com

The present study was supported by grants from Kidney Research UK (ST4/2011), the Cunningham Trust (CT11/14) and the Mrs EA Hogg's Charitable Trust.

注:动物实验已根据由动物（科学规程）1986年法令所规定的准则和法规执行的程序使用无菌（高温高压）手术器械和耗材进行。 15周，最适年龄感 - 虽然肾IRI的小鼠模型这里介绍的是在8周龄雄性它可以在各种不论男女通常7岁之间的小鼠品系的可再现进行的BALB / c小鼠进行共8周。在代表性的结果部分给出的数据是来自BALB / c和FVB小鼠获得的。温热生理盐水的应用是用来保持肠内和手术区域湿润，但它应仔细监测并保持在最低水平作为流体的过度应用可导致伪迹降低血清或血浆肌酸酐水平，这是一个重要的实验读数。 1，动物的制备与剖腹手术 &lt;醇&gt; <li>麻醉小鼠用氯胺酮盐酸盐（70毫克/千克）和盐酸美托咪定（1毫克/千克）腹膜内和由反射例如脚趾捏损失确认麻醉深度。所得到的麻醉平面的持续时间是4小时。这足以完成整个手术过程并没有补充麻醉是必需的。 </li><li>除去周围切口区头发上，确保该地区是免费的松散的头发，并用消毒稀洗必泰溶液的腹部皮肤。 </li><li>将鼠标上的外科加热垫的仰卧位置，并使用低粘性胶带固定在上肢和下肢的垫。确保上肢都保持在正常的位置，以防止肺压缩。整个手术过程中监控鼠标的热灼伤。如果可能的话，建议非电动热源使用。 </li><li>施用的镇痛盐酸丁丙诺啡（0.06毫克/千克）皮下和敷眼润滑剂涂抹到眼睛，以防止角膜干燥。止痛药是为了帮助手术后的恢复管理的预操作。 </li><li>采用组织分离剪刀或手术刀刀片做中线剖腹切口直言分离腹膜皮肤。这使得皮肤和腹膜待伤口愈合过程中分别缝合。无血管白线的切口，使进入腹膜腔。 </li><li>要建立手术区的清晰视图中插入一个拉钩，悬垂鼠标。 </li></ol> 2，输尿管司和右肾切除<ol><li>轻轻推入肠对使用蘸有生理盐水，暴露右肾及输尿管高压灭菌消毒棉签腹腔左侧。盖上湿润的布肠道，防止干燥。 </li><li>抬起右输尿管与角度的钳。结扎右输尿管采用6/0 SIL两次K-编织缝合。对于长期的实验中，使用可吸收缝合线的所有腹部手术。 </li><li>划分缝线之间的输尿管。虽然膀胱输尿管交界处，应防止尿液漏入腹膜从膀胱输尿管被结扎作为对在长期实验中泄漏任何手术后一种保障。 </li><li>为了使用户更容易访问进行右肾切除时，用沾湿的棉签轻轻推肝脏向上和向右并固定到位用湿润的纱布，暴露右肾动脉和静脉。 </li><li>说白了解剖结缔组织和脂肪沿右肾内侧方面尽可能的右肾动脉和静脉。 </li><li>建立肾动脉和静脉通过仔细滑动倾斜的镊子血管下方下方的通道。引导钳子下方的提示关闭，并轻轻地去除与提示，以便于信道的形成开路。</li><li>重复步骤2.6直到钳提示是可见的只是通过肾动脉和静脉上方的结缔组织。一旦可见轻轻揉搓钳提示对另一套角度的钳的尖端轻轻地打破结缔组织。 </li><li>与肾动脉和静脉明显地可访问他们现在可以遮挡。小心地将倾斜的镊子肾动脉下方，静脉与提示关闭。一旦到位开镊子的提示和引导技巧之间的止血夹子涂药，牢固应用止血夹到肾动脉和静脉靠近肾脏。另外肾动脉和静脉可以使用6/0丝线编织缝线结扎来。 </li><li>划分闭塞肾动脉和静脉靠近肾脏，这现在可以与任何剩余的贴壁结缔组织中删除。 </li><li>卸下用来保存肝脏和用棉签代替肠子任何纱布。 </li></ol><ol><li>轻轻推肠子朝右侧腹腔棉签暴露左肾及输尿管，并盖上湿润的毛巾。如果有必要的胰腺可以与湿润纱布被偏转，以允许更容易获得。 </li><li>使用钝性分离轻轻打破结缔组织前部和后部的左侧肾动脉和静脉，然后创建以类似的方式船只下方的通道，在输尿管司和右肾切除2.4描述的步骤。 </li><li>通过应用微serrafine夹到肾动脉和静脉引起左肾缺血。成功的缺血可以通过肾脏逐渐变黑统一进行目视确认。血管除了左肾动脉和静脉偶尔可以提供肾脏。如果存在的话，这些额外的血管还需要使用微serrafine片段，以便成功地被遮挡导致整个肾脏的缺血。 </li><li>更换肠子和再三保证，没有突然的曲折，可能会导致肠缺血妥协的结果。暂时与单个缝合关闭腹膜。 </li><li>局部缺血诱导马上将鼠标放在一个恒温动物毯与连接到一个控制单元，其将维持体温在37℃缺血的所需持续时间的直肠热敏电阻。定义必要缺血的适当长度来诱导的肾损伤和肾功能衰竭的期望程度，建议将每个菌株和手术操作者进行滴定。 </li><li>不久缺血期结束前重新打开腹膜，仔细定位肠子允许访问钳和查看肾脏。卷收器的插入，如视频所示，是没有必要的，是专为表象的目的而进行的。 </li><li>取出CL缺血期后放的结论。立刻取出后，在肾脏会迅速从一个深栗色改变颜色，以一个健康的深粉红色，表示成功的再灌注。 </li><li>再次确保肠子不关闭腹膜使用6/0编织丝线缝合一针毯之前，扭曲。然后关闭使用金​​属夹子皮肤的皮肤。 </li><li>为了尽量减少术后感染的风险，应用防腐剂如碘/酒精溶液至手术区。 </li></ol> 4，术后恢复和护理<ol><li>部分反向麻醉阿替美唑盐酸盐（2毫克/千克）经皮下给药和流体用皮下注射1毫升温热生理盐水，以防止脱水以下手术。 </li><li>仔细监测小鼠，直到他们恢复了意识，出现警报，并能正确自己。 </li><li>允许小鼠在加热箱恢复保持在29℃，禄ATED在一个安静的环境，24小时。小鼠将不得不温度调节因麻醉的能力受损，因此，至关重要的是，它们被安置在升高的温度，以使有效的恢复。湿润的食物也可以提供，以鼓励液和营养摄入。 </li><li>对于长期回收实验提供持续的止痛药和手术去除皮肤剪辑7天。 </li></ol> 5，评估功能和结构肾损伤和再生<ol><li>血可从尾静脉在实验过程中或通过心脏穿刺来收集在安乐死的时间在实验结束。通过血清肌酸酐测量肾功能，使用于生化分析仪离心一个基于肌酸方法，以及血液尿素氮（BUN），如先前描述的7。需要正常健康小鼠的肾功能建立肾功能衰竭的模型，因为这之前确定可以改变明显dependi纳克在使用如谢斐反应，并根据肌酸方法测量肌酐的分析方法给出不同的结果。 </li><li>经病理组织学上的石蜡包埋肾组织切片用苏木精和曙红（H＆E）或碘酸雪夫（PAS）检查肾功能结构。在200倍放大率的外髓（OSOM）对每只小鼠外条纹内10幅图像 - 拍摄之间5。评估在OSOM损伤的程度，因为这地区有伤在这个模型中，因为它是非常容易受到缺氧。 </li><li>请使用下面详述两个评分系统来衡量急性肾小管坏死（ATN）或再生。在这些系统中使用的分类的代表的形态示于图1。 </li><li>用一个简单的二进位制的得分ATN。对细胞的完整性和形态标记的基础和计数肾小管无论是作为可行的（完整的细胞形态）或坏死（细胞受损集成grity，细胞形态异常或细胞的损失）。 <ol><li>表达肾小管坏死的数量作为细管的总数（坏死的肾小管％）的百分比。 </li></ol></li><li>使用基于细胞的完整性，形态及核数另一个系统分类再生。马克和计数肾小管一样健康，坏死，受伤或按照以下步骤列出的标准中恢复过来。 <ol><li>由于与ATN评分系统（在步骤5.4详述）肾小管标志与完好正常细胞的健康，而小管出妥协细胞的完整性，细胞形态异常或明显的细胞损失应评价为坏死。 </li><li>分类小管和受伤，如果他们有一个包含几个原子核的薄壁细胞质中。相比之下，指定含有较多的细胞核较正常的细胞形态为肾小管恢复。 </li><li>表示每个小管分类为总肾小管数的百分比。 </li></ol></li></ol>

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The authors have no competing or conflicting interests to disclose.

肾IRI是AKI的一个重要原因，没有提供具体的治疗。肾脏IRI的实验研究已经在以前的工作表明巨噬细胞，树突细胞，淋巴细胞，调节性T细胞以及其他细胞和两者的急性损伤和愈合阶段5,8中感应介质的作用非常翔实- 16。此外，实验肾脏IRI已被用于评估各种治疗剂4,17-19的效果。 肾IRI这里详述的模型使用一个中线剖腹手术的方法来进行右肾切除，并用夹子?...

缺血再灌注损伤（IRI）是受伤的多个器官，包括肾脏，心脏和大脑的共同模式。肾IRI可能导致急性肾损伤（AKI）患者，并没有具体的治疗方法。 AKI作为IRI的结果具有复杂的发病机理涉及两个先天和适应性免疫应答1。肾IRI的实验模型提供解剖的关键细胞，并参与了AKI的发病机制，以及随后的肾再生随之而来的是在随后的日子里调解员的潜力。此外后疾病进程的新的治疗剂的效果进行评估。 这里所描述的肾IRI的实验模型的总体目标是诱导急性功能和结构的肾损伤。一些研究者已经利用涉及双边IRI 2的诱导的模型。虽然双侧肾IRI模型的使用，单方面仁人IRI模型有一个右肾切除正在开展在手术时间的优势。右侧肾切除的组织可作为有价值的对照组织中的研究涉及要么诱导或抑制的基因或蛋白质的表达的预处理步骤。例如，我们用这个模型来评估血红素精氨酸的预处理效果（HA）注射肾IRI 3 24小时前交回。成功诱导细胞保护酶血红素加氧酶-1（HO-1）由医管局IRI之前在右肾切除对照组织4中得到证实。房委会通过的HO-1依赖的机制减少肾IRI在老年小鼠中的一部分。类似地，我们已经使用了模型中巨噬细胞的耗竭研究，探讨巨噬细胞在肾IRI 5的作用。右侧肾切除组织的免疫组织化学分析可用于确定消融方法的疗效。右侧肾切除的组织，因此可以用来同时确认和量化i的平nduction或抑制在每个个体实验动物感兴趣分子的。这个模型将是感兴趣的谁正在使用的药物或其他化合物来调节肾IRI的诱导前基因或蛋白质等的表达研究。 其他研究已经使用侧面切口访问肾脏。这里所描述的模型使用一个单一的中线腹部手术同时执行的右肾切除，诱导左肾缺血再灌注损伤。这种手术方式提供了出色的外科领域，包括肾蒂和颜色的变化遵循肾蒂夹紧的可视化。我们的出版经验与模型4-6表明，小鼠很快从手术中恢复了接近100％的存活率。 最后，该模型在一个为期7天的动力学分析表明，该模型具有两个恢复肾功能和肾小管完整性，用显著肾小管细胞增殖。

<table><tbody><tr><td>Tissue scissors</td><td>Fine Science Tools</td><td>14072 - 10</td><td></td></tr><tr><td>Micro-Adson forceps (Rat toothed)</td><td>Fine Science Tools</td><td>11019 - 12</td><td></td></tr><tr><td>S&amp;amp;T JFA-5bTC Forceps - SuperGrip Angled</td><td>Fine Science Tools</td><td>00649-11</td><td></td></tr><tr><td>Colibri retractor</td><td>Fine Science Tools</td><td>17000 - 04</td><td></td></tr><tr><td>Micro clip applicator</td><td>Fine Science Tools</td><td>18057-14</td><td></td></tr><tr><td>Micro serrafines&amp;nbsp;</td><td>Fine Science Tools</td><td>18055-04</td><td></td></tr><tr><td>Olsen-Hegar needle holder</td><td>Fine Science Tools</td><td>12002 - 12</td><td></td></tr><tr><td>Hemoclip Plus Ligating Clips Small</td><td>Weck</td><td>533837</td><td></td></tr><tr><td>Autoclip Wound Clip System, 9mm</td><td>Harvard Apparatus</td><td>PY2 52-3748</td><td></td></tr><tr><td>Silk Black Braided Suture, Size 6-0</td><td>Harvard Apparatus</td><td>723288</td><td></td></tr><tr><td>Standard Heat Matt</td><td></td><td></td><td></td></tr><tr><td>Homeothermic Blanket &amp;amp; Control Unit</td><td>Harvard Apparatus</td><td></td><td></td></tr><tr><td>Lacri-Lube</td><td>Allergan</td><td></td><td></td></tr><tr><td>Vetasept Chlorhexidine&amp;nbsp;&amp;nbsp;</td><td>AnimalCare</td><td></td><td></td></tr><tr><td>Vetalar : Ketamine hydrochloride</td><td></td><td></td><td>100 mg/ml solution</td></tr><tr><td>Domitor : medetomidine hydrochloride&amp;nbsp;</td><td></td><td></td><td>1 mg/ml</td></tr><tr><td>Vetergesic : Buprenorphine hydrochloride&amp;nbsp;</td><td></td><td></td><td>0.3 mg/ml</td></tr><tr><td>Antisedan : Atipamezole hydrochoride</td><td></td><td></td><td>5 mg/ml</td></tr></tbody></table>

renal ischaemia reperfusion injury: a mouse model of injury and regeneration

肾移植术中肾缺血再灌注损伤（IRI）是急性肾损伤（AKI）患者和肾血流量的阻塞的常见原因是不可避免的。实验模型，准确和可重复地概括肾IRI在解剖AKI的病理生理和新的治疗药物的发展是至关重要的。这里介绍的是肾IRI小鼠模型，结果在重复性的AKI。这是通过对与一个切口允许两个右肾切除术，它提供控制组织和左肾蒂夹紧诱导左肾缺血的手术中线剖腹手术的方法来实现。通过在缺血期间仔细监测夹紧位置和体温的这种模式实现了可重复的功能和结构的损伤。小鼠处死24小时以下手术示范肾功能的丧失与血清或血浆肌酸酐水平的升高，以及结构郭晓丽肾损害与急性肾小管坏死明显。肾功能改善和肾IRI这样，该模型可用于研究肾再生7天期间的急性组织损伤消退。已动用肾IRI的这个模型，研究AKI的分子和细胞病理生理学，以及随后的肾再生的分析。

肾小管损伤和恢复可以由H＆E或评估的组织切片肾IRI PAS染色。位于OSOM内小管是根据细胞形态，完整性和核数（ 图1）划分为健康，受伤，坏死或恢复。在此模型中的功能和结构的损伤是依赖于缺血的持续时间。逐步增加肾功能不全的严重程度，由血浆肌酐和尿素氮评估，是明显的，因为缺血的持续时间增加2分钟的增量（ 图2）。结构肾损伤，从ATN得分推断的程度，如下更...

Watch this Scientific Journal Video about Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration at JoVE.com

Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration

The mouse model of renal ischaemia reperfusion injury described here comprises of a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia that results in acute kidney injury. This model uses a midline laparotomy approach with all steps performed via one incision.

肾缺血再灌注损伤:损伤的小鼠模型和再生

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable ...

renal-ischaemia-reperfusion-injury-mouse-model-injury

Research

JoVE Journal

Medicine

49.6K 次观看.  University of Edinburgh. The mouse model of renal ischaemia reperfusion injury described here comprises of a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia that results in acute kidney injury. This model uses a midline laparotomy approach with all steps performed via one incision.

视频: Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration

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

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 ...

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, ...

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 ...

Technical Refinement of a Bilateral Renal Ischemia-Reperfusion Mouse Model for Acute Kidney Injury Research

Cardiac arrest poses a large public health burden. Acute kidney injury (AKI) is an adverse marker in survivors of cardiac arrest following the return of spontaneous circulation (ROSC) after successful cardiopulmonary resuscitation. Conversely, recovery of kidney function from AKI is a predictor of favorable neurological outcomes and hospital discharge. However, an effective intervention to prevent kidney damage caused by cardiac arrest after ROSC is lacking, suggesting that additional therapeutic strategies are required. Renal hypoperfusion and reperfusion are two pathophysiological mechanisms that cause AKI after cardiac arrest. Animal models of ischemia-reperfusion-induced AKI (IR-AKI) of both kidneys are comparable with patients with AKI following ROSC in a clinical setting. However, IR-AKI of both kidneys is technically challenging to analyze because the model is associated with high mortality and wide variation in kidney damage, which may affect the analysis. Lightweight mice were chosen, placed under general anesthesia with isoflurane, subjected to surgery with a dorsolateral approach, and their body temperature maintained during operation, thereby reducing tissue damage and establishing a reproducible acute renal IR-AKI research protocol.

This study established a protocol focusing on the technical refinement of a mouse model of bilateral renal ischemia-reperfusion for acute kidney injury research.

用于急性肾损伤研究的双侧肾缺血再灌注小鼠模型的技术改进

Cardiac arrest poses a large public health burden. Acute kidney injury (AKI) is an adverse marker in survivors of cardiac arrest following the return of ...

Developing Murine Acute Kidney Injury Model Using Bilateral Ischemia-Reperfusion

Bilateral Renal Ischemia-Reperfusion Model for Acute Kidney Injury in Mice

Acute kidney injury (AKI) is defined as a rapid decline in renal function, in which persistent kidney dysfunction gradually progresses to chronic kidney disease (CKD) due to the irreversible loss of nephrons and their maladaptive repair. In recent years, the incidence of AKI has been increasing concerning diverse etiologies, including volume depletion, sepsis, nephrotoxicity, muscle injury, and major trauma, in which ischemia-reperfusion injury (IRI)&#160;accounts for most episodes. Development of the IRI model in mice is induced by surgical clamping of the renal pedicles, which provides powerful and controllable tools for preclinical models of AKI. Importantly, the IRI model is deployed at different stages of the AKI development, especially in the processes of AKI to CKD. Despite the IRI model being widely practiced in many laboratories, a series of variables still influence the results of this model. Here, we describe the procedure of IRI model development to provide a repeatable and reliable method for researchers to explore the underlying pathogenesis in the development of AKI and the progression of AKI to CKD.

The ischemia-reperfusion injury (IRI) model can be used at different stages of acute kidney injury (AKI) development, especially during AKI to chronic kidney disease (CKD) progression. Here, we describe the procedure for IRI model development in mice via a trans-abdominal approach, clamping renal pedicles via a vascular clip to induce ischemic injury.

Acute kidney injury (AKI) is defined as a rapid decline in renal function, in which persistent kidney dysfunction gradually progresses to chronic kidney ...

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.