Name: induction of myocardial infarction and myocardial ischemia-reperfusion injury in mice
Uploaded: 2022-01-19T13:30:00
Description: Watch this Scientific Journal Video about Induction of Myocardial Infarction and Myocardial Ischemia-Reperfusion Injury in Mice at JoVE.com

这项工作得到了中国国家自然科学基金（82070317,81700390林继斌，8210021880吕冰杰，82000428王博元）和国家重点研发计划（2017YFA0208000给何少林）的支持。

动物研究已获得华中科技大学（中国武汉）动物护理和利用委员会的批准。
注：雄性C57BL / 6J小鼠（8-10周）用作模型。小鼠可以自由获取食物和水，并在特定的无病原体条件下繁殖。房间保持在受控温度（22°C±2°C）和湿度（45%-65%）下。根据该机构制定的指南，将小鼠暴露在同济医学院（中国武汉）动物护理设施的12小时光/暗环境中。使用无菌显微手术器械和手术用品。在整...

<ol>
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近年来，MI和MIRI模型在临床和科学研究中的创建发展迅速20,21。然而，仍有一些问题需要解决，例如作用机制和如何改善MI/MIRI。在这里，描述了用于建立MI和MIRI小鼠模型的修改方案。必须仔细考虑几个关键点。
第一个关键点是气管插管。一些程序6,9 涉及切开颈椎皮肤、组织分离，然后暴...

急性心肌梗死 （AMI） 是全球常见的心血管疾病，死亡率很高1.技术的进步使AMI患者可以进行早期有效的血运重建。在一些患者进行这些治疗后，可能会发生心肌缺血再灌注损伤 （MIRI）2。因此，了解MI/MIRI的作用机制和改善MI/MIRI具有重要意义。小鼠因其成本低、繁殖时间短且易于进行基因改变而被广泛用作模型3.学者们开发了不同的方法来模拟动物4、5、6、7、8、9 的 MIRI 和 MI。这种策略促进了研究，但采用的不同标准和方法使研究团队对结果的解释变得复杂。
在小鼠中，异丙肾上腺素 10、冷冻损伤11,12 或烧灼13 诱导了心肌梗死。异丙肾上腺素易诱发心肌梗死，但病理生理过程与临床心肌梗死不同。 冷冻损伤诱发的心肌梗死一致性差，引起左冠状动脉前降支（left anterior lower coronary artery，LAD）周围过度心肌损伤，易诱发心律失常。烧灼诱发的心肌梗死与心肌梗死的自然过程有很大不同，烧灼部位的炎症反应更强烈;此外，手术方法存在技术困难。此外，还有一些实验室14 通过介入技术使用球囊阻塞或栓塞或血栓形成方法在小型猪中开发 MI 模型。所有这些方法都可能直接导致冠状动脉闭塞，但需要冠状动脉造影设备，尤其是太薄的小鼠冠状动脉使这些手术不切实际。对于MIRI，不同模型之间的差异非常小，例如是否使用呼吸器/显微操作5,6。
在这里，描述了一种简单可靠的方法可以诱导MI和MIRI模型，该模型改编自先前发表的方法4,5,6,7,8,9,15。该方法可以通过结扎直接阻断LAD来模拟病理生理过程。此外，通过解除结扎，该模型还可以模拟再灌注损伤。在该协议中，解剖显微镜用于LAD可视化。然后，研究人员可以很容易地识别 LAD。随后，LAD的准确结扎导致可重复和可预测的血液闭塞和心室缺血。此外，除了在显微镜下观察到的 LAD 颜色变化外，心电图 （ECG） 变化还可用于确认缺血和再灌注。这种策略导致更短的手术时间，更低的手术并发症风险，以及更少的实验小鼠。还描述了肌钙蛋白-T 试验、心脏超声和氯化三苯基四唑 （TTC） 染色的方法。总体而言，该方案可用于MI / MIR机制的研究以及药物发现。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9 % sodium chloride solution</td>
			<td>Kelun Industry Group,China</td>
			<td></td>
			<td>-</td>
		</tr>
		<tr>
			<td>4% paraformaldehyde fixing solution</td>
			<td>Servicebio,China</td>
			<td>G1101</td>
			<td>-</td>
		</tr>
		<tr>
			<td>4-0 silk suture</td>
			<td>Shanghai Pudong Jinhuan Medical Products,China</td>
			<td>C412</td>
			<td>-</td>
		</tr>
		<tr>
			<td>8-0 suture</td>
			<td>Shanghai Pudong Jinhuan Medical Products,China</td>
			<td>H801</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>IsoReag,China</td>
			<td>IR-11190</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Camera</td>
			<td>Canon,Japan</td>
			<td>EOS 80D</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Depilatory cream</td>
			<td>Veet,French</td>
			<td></td>
			<td>-</td>
		</tr>
		<tr>
			<td>Elecsys Troponin T hs STAT</td>
			<td>Roche,Germany</td>
			<td></td>
			<td>-</td>
		</tr>
		<tr>
			<td>Electrochemical luminescence immunoanalyzer</td>
			<td>Roche,Germany</td>
			<td>Elecsys 2010</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Evans blue</td>
			<td>Sigma,America</td>
			<td>E2129</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Eye scissors</td>
			<td>Shanghai Medical Instruments,China</td>
			<td>JC2303</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Haemostatic forceps</td>
			<td>Shanghai Medical Instruments,China</td>
			<td>J31020</td>
			<td>-</td>
		</tr>
		<tr>
			<td>High frequency in vivo imaging systems</td>
			<td>Visualsonics,Canada</td>
			<td>Vevo2100</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Ibuprofen</td>
			<td>PerFeMiKer,China</td>
			<td>CLS-12921</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Intravenous catheter</td>
			<td>Introcan,Germany</td>
			<td>4254090B</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Sigma-Aldrich,America</td>
			<td>&#160;K2753</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Medical alcohol</td>
			<td>Huichang ,China</td>
			<td></td>
			<td>-</td>
		</tr>
		<tr>
			<td>Microneedle holders</td>
			<td>Shanghai Medical Instruments,China</td>
			<td>WA2040</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Microscopic shears</td>
			<td>Shanghai Medical Instruments,China</td>
			<td>WA1040</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Microsurgical forceps</td>
			<td>Shanghai Medical Instruments,China</td>
			<td>WA3020</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Mouse electrocardiograph</td>
			<td>Techman,China</td>
			<td>BL-420F</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Needle holders</td>
			<td>Shanghai Medical Instruments,China</td>
			<td>JC3202</td>
			<td>-</td>
		</tr>
		<tr>
			<td>operating floor</td>
			<td>Chico,China</td>
			<td>ZK-HJPT</td>
			<td>-</td>
		</tr>
		<tr>
			<td>PE-10 tube</td>
			<td>Huamei,China</td>
			<td></td>
			<td>-</td>
		</tr>
		<tr>
			<td>Pentobarbital</td>
			<td>Merck,America</td>
			<td>1030001</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Rodent Ventilator</td>
			<td>Shanghai Alcott Biotech,China</td>
			<td>ALC-V8S-P</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Stereo microscope</td>
			<td>Aomei Industry,China</td>
			<td>SZM0745-STL3-T3</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Surgical thermostatic heating pad</td>
			<td>Globalebio, China</td>
			<td>GE0-20W</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Triphenyltetrazolium chloride</td>
			<td>Servicebio,China</td>
			<td>G1017</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Huamaike Biochemicals and Life Science Research Prouducts,China</td>
			<td>323004</td>
			<td>-</td>
		</tr>
	</tbody>
</table>

induction of myocardial infarction and myocardial ischemia-reperfusion injury in mice

急性心肌梗死是一种常见的心血管疾病，死亡率很高。心肌再灌注损伤可以抵消心脏回流的有益作用，诱发继发性心肌损伤。简单且可重复的心肌梗死和心肌缺血再灌注损伤模型是研究人员的良好工具。在这里，描述了一种通过显微操作精确结扎左前降支冠状动脉 （LAD） 来创建心肌梗死 （MI） 模型和 MIRI 的可定制方法。LAD 的准确且可重复的结扎定位有助于获得一致的心脏损伤结果。ST 段更改有助于识别模型准确性。血清心肌肌钙蛋白T（cTnT）水平用于评估心肌损伤，心脏超声用于评估心肌收缩功能，Evans-Blue/三苯基氯化四氮唑染色用于测量梗死面积。一般来说，该协议减少了手术时间，确保了可控的梗死大小，并提高了小鼠的存活率。

实验工作流程如图 1A所示。研究开始后，研究者可以根据实验设计安排时间节点。LAD结扎的持续时间取决于研究目的。对于心肌梗死，研究可以忽略再灌注步骤。心脏超声可在研究的不同阶段进行，因为它是非侵入性的，而 Evans-Blue/TTC 染色只能在小鼠被处死时进行。对于专注于纤维化和心室重塑的研究，观察时间要长得多。
部分实验过程的典型图像如?...

Watch this Scientific Journal Video about Induction of Myocardial Infarction and Myocardial Ischemia-Reperfusion Injury in Mice at JoVE.com

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

在这里，我们描述了一种简单且可重复的方法，该方法可以通过显微操作精确结扎左前降支冠状动脉来诱导小鼠心肌梗死或心肌缺血再灌注损伤。

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 ...

本研究描述了一种可定制的方法，通过显微操作精确结扎左前降支冠状动脉，在 Miro 中创建心肌功能模型。该协议减少了手术时间，确保了可控的梗死大小，并提高了小鼠的存活率。在整个手术过程中，使用带有预热至 37 摄氏度的加热垫的矩形 OT。手术前，用紫外线和70%酒精对电路板进行消毒。首先将鼠标放在 OT 上，仰卧位，头部下方有纱布，以免眼睛过热。将眼药膏涂抹在眼睛上，以防止眼睛干燥。在胸部使用毛发去除霜，用无菌棉签均匀按摩一分钟。用纱布擦拭多余的毛皮。使用聚维酮碘，然后使用70%酒精清洁该区域，并用纱布覆盖胸部。在上切牙下方使用 4-O 缝合线并将其固定在锚点上，以保持嘴巴略微张开并便于插管。拉动尾巴以保持身体笔直，并用胶带将其固定在 OT 上。用胶带固定四肢并将它们拧紧在锚点上。不要过度伸展前肢，以免呼吸功能受损。用镊子打开下颌并抬起舌头。然后使用照明器观察喉咙和声门。轻轻地将一根 22 号插管和一根钝的截短针穿过口腔插入气管，在喉咙下方约 1 厘米处。用一只手握住舌头，用钝镊子稍微向上移动。同时用另一只手将管子轻轻插入气管。小心不要将管子插入食道。小心地取下针头。在连接到呼吸机之前，通过将管子放入水中以形成气泡来检查插管。将气管插管连接到设置为每分钟 120 次的呼吸机，并将潮气量调节为 250 微升。通过检查双侧对称性胸廓扩张来验证插管。然后用胶带固定与 OT 的连接，以免其脱落。将心电图电极放在爪子上并将它们连接到心电图记录仪。在整个手术过程中监测心脏电生理学。取下胸部的纱布。用 70% 酒精对切口区域进行三次擦洗循环消毒。为了减少手术部位的污染，用无菌手术单覆盖小鼠，在手术区域上有一个孔。用手术刀做斜皮肤切口后，通过对皮下组织进行钝性解剖来暴露肋骨。注意不要伤害血管、肋骨和肺部。使用无菌棉涂抹器止血。确定第三肋间隙，切开约 6 至 8 毫米的切口。然后通过对肋间隙中的组织进行钝性解剖来打开胸腔。注意不要伤到胸内动脉。使用镊子跨越肋间隙。将预先消毒的自制牵开器插入胸腔，然后向后拉，将切口扩散到大约六毫米宽。用橡皮筋将牵开器连接到 OT 上。小心地切除周围的组织，使心脏完全暴露。用弯曲的镊子轻轻拉出心包，而不会伤害心脏。使用解剖显微镜并引导聚焦且适当的光线进行 LAD 可视化。轻轻按压所选结扎位置下方的部位以暂时扩大 LAD，然后重新检查 LAD。在解剖显微镜下，使用锥形针通过 8-0LAD下方的丝绸结扎。确保针头不够深而无法进入左心室，也不要太浅，以免损坏 LAD。用松散的双结系紧结扎。将一根 2 到 3 毫米的 PE-10 管放入平行于动脉的环中。轻轻收紧结扎环，直到它绕过动脉和管道。然后用滑结固定环。注意不要因过度收紧压力而损坏心肌壁。结扎后左心室前壁苍白，证实LAD血流停止。如果需要永久结扎，请按照文本手稿中的说明进行。从切口上取下牵开器。然后用斗牛犬夹暂时闭合伤口。确保鼠标继续连接到呼吸机。当缺血期结束时，取下斗牛犬夹并插入牵开器以打开切口并暴露心脏和结扎部位。解开滑结并取下 PE-10 管。苍白的颜色变为粉红色，证实了血流恢复。同时，为了检测再灌注，观察心电图是否有ST段抬高的潜在溶解。离开 8-0如果进行随后的埃文斯蓝和 TTC 染色，则原位结扎。否则，请在此步骤中移除缝合线。取出牵开器，用4-O丝线缝合第三和第四肋骨，闭合切口。注意不要伤到肺部。轻轻按压胸部，同时系上缝合结，以排出可能滞留在胸腔中的空气。用连续的4-O丝线缝合肌肉层。从气管插管、皮肤切开、开胸手术、LAD鉴定和结扎到再灌注，均获得实验过程的标准图像。代表性心电图图像证实了结扎后心肌缺血和再灌注，并伴有显著的ST段抬高。滑结解开后ST段抬高的溶解。开胸手术前，保持气道畅通非常重要，最重要的是确保LAD结扎牢固，避免损伤。该程序之后将进行超声心动图以评估心脏功能以及 TTC 以评估心肌损伤。

Research

JoVE Journal

Medicine

Acute Myocardial Infarction in Rats

大鼠急性心肌梗死

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular ...

科研

医学

Induction of Myocardial Infarction in Adult Zebrafish Using Cryoinjury

在成年斑马鱼心肌梗死的感应使用Cryoinjury

The mammalian heart is incapable of significant regeneration following an acute injury such as myocardial infarction1. By contrast, urodele amphibians and ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

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 ...

MRI and PET in Mouse Models of Myocardial Infarction

MRI和PET心肌梗死小鼠模型

Myocardial infarction is one of the leading causes of death in the Western world. The similarity of the mouse heart to the human heart has made it an ...

Myocardial Infarction and Functional Outcome Assessment in Pigs

心肌梗死和功能预后评估的猪

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One ...

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

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 ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

在诱导小鼠心肌梗死的微创手术过程

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

诱导和缺血再灌注损伤评估中的Langendorff灌注鼠心

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

Myocardial Infarction in Neonatal Mice, A Model of Cardiac Regeneration

心肌梗死新生小鼠，心肌再生的模型

Myocardial infarction induced by coronary artery ligation has been used in many animal models as a tool to study the mechanisms of cardiac repair and ...

Confirmation of Myocardial Ischemia and Reperfusion Injury in Mice Using Surface Pad Electrocardiography

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 ...