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 border="0" cellpadding="0" cellspacing="0" width="1160">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Heated Pad</td>
			<td style="width:191px;">Sunbeam</td>
			<td style="width:249px;">E12107-819</td>
			<td style="width:379px;">Alternative: Braintree Scientific heated pad</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Table top research anesthesia Machine</td>
			<td>Vasco</td>
			<td>UCAP 0001-0000171</td>
			<td>Alternative: Parkland Scientific, V3000PS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nose Cone</td>
			<td>Parkland Scientific</td>
			<td>ARES500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scavenger canister and replacement cartridge</td>
			<td>Parkland Scientific</td>
			<td>80000, 80120</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Induction Chamber</td>
			<td>Surgivet</td>
			<td>V711802</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Piramal Healthcare</td>
			<td>NDC 66794-013-10</td>
			<td>Controlled substance, contact IACUC</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Animal clipper&#160;</td>
			<td>Oster&#160;</td>
			<td>Oster Golden A5 078005-050-003</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ophthalmic ointment</td>
			<td>Webster</td>
			<td>8804604</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenorphine</td>
			<td>McKesson</td>
			<td>562766</td>
			<td>Controlled substance,contact IACUC</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ketaset (Ketamine HCl)</td>
			<td>Pfizer</td>
			<td>NADA 45-290</td>
			<td>Controlled substance, contact IACUC</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cotton tips</td>
			<td>Puritan medical products</td>
			<td>806-WC</td>
			<td>Autoclave before use</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Betadine</td>
			<td>Purdue Products</td>
			<td>67618-150-17</td>
			<td>10% Povidone-Iodine</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile saline solution</td>
			<td>Aspen</td>
			<td>46066-807-60</td>
			<td>Adjust to room temperature before use</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile cotton gauze pad</td>
			<td>Fisher Healthcare</td>
			<td>22-415-468</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Non-adherent pad prepack</td>
			<td>Telfa</td>
			<td>1238</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">IR rodent thermometer</td>
			<td>BIOSEB</td>
			<td>BIO-IRB153</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro vascular clips, 70g</td>
			<td>Roboz Surgical&#160;</td>
			<td>RS5424, RS5435</td>
			<td>Alternative: WPI 14121, for SMA occlusion</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro vascular clips, 40g</td>
			<td>Roboz Surgical&#160;</td>
			<td>RS6472</td>
			<td>Alternative:WPI 14120, for collateral vessels occlusion</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Clip applying forceps</td>
			<td>World Precision Instruments</td>
			<td>14189</td>
			<td>Alternative: Roboz #RS-5410 or&#160; #RS-5440</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gill&#39;s 3 hematoxylin</td>
			<td>Thermo Scientific</td>
			<td>14-390-17</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical staples, Reflex 9 mm</td>
			<td>Cell Point Scientific</td>
			<td>201-1000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Autoclip applier</td>
			<td>Beckton Dickinson</td>
			<td>427630</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Byopsy foam pad</td>
			<td>Simport</td>
			<td>M476-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tissue cassette</td>
			<td>Fisher Healthcare</td>
			<td>15182701A</td>
			<td>Histosette II combination lid and base</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10% buffered formalin</td>
			<td>Fisher Scientific</td>
			<td>245-684</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical iris scissors</td>
			<td>World Precision Instruments</td>
			<td>501263-G SC</td>
			<td>Alternative: Roboz RS6816</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Operating scissors</td>
			<td>World Precision Instruments</td>
			<td>501219-G</td>
			<td>Alternative: Roboz RS6814</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dressing forceps</td>
			<td>Roboz Surgical&#160;</td>
			<td>RS-5228, RS-8122</td>
			<td>Alternative: World Precision Instruments 1519-G</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Heparin, endotoxin free, 300 USP units/vial, 50mg</td>
			<td>Sigma</td>
			<td>2106</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:343px;">Reflex wound clip removing forceps</td>
			<td>Roboz Surgical&#160;</td>
			<td>RS-9263</td>
			<td>Alternative: World Precision Instruments: 500347</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Mice C57BL/6J mice&#160;</td>
			<td>Jackson Laboratory</td>
			<td>Stock No 0664</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Telfa non-adherent dressings, 3x4, sterile</td>
			<td>Coviden</td>
			<td>1050</td>
			<td></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

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

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 ...

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, ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

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

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

大鼠心肌缺血再灌注后冠状动脉流动储备评价

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

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

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

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

重建显微外科缺血再灌注损伤研究的临床前大鼠模型

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

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

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

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

在穆林模型中缺血再灌注损伤后干细胞的心肌内传递延迟

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

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

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

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

器官缺血-输液损伤模拟大鼠肝脏移植模型的血动力学变化

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

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 ...

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 ...