的BKM是一个美国国立卫生研究所（NIH）/国家心肺和血液研究所（NHLBI）的补助金（R01 - HL085487）的支持。 MH是由美国心脏协会（AHA）的国家科学家开发格兰特（SDG）和老年心脏病学协会的支持。

1。经营面积的制备<ol><li>作业面是先准备消毒用70％乙醇面积。 </li><li>所有手术器械消毒珠手术前用热灭菌器（和个人之间的大鼠手术）。这些文书包括：（2）手术剪，钳（1），弯钳（1），持针器（2），和胸部rectractor。 </li><li>棉花，纱布和施药，可用以管理出血。 </li><li>调整温度的一个homeothermic毯系统（电热垫），用于动物的体温保持在37 ± 1℃，以防止体温急剧下降;梗死的大小是依赖于闭塞以及体温（Tarnavski 等 ）的持续时间。 </li><li>麻醉前，每个动物将接受手术前一天的剂量丁丙诺啡（0.1-2.5毫克/公斤SQ）和卡洛芬口服片剂（Rimadyl，Bioserv片剂1片）。当时立即手术之前，每个动物将再次收到剂量丁丙诺啡（0.1-2.5毫克/公斤SQ）。 </li><li>注：对于涉及过了一段时间的评估心脏功能和心脏重塑的研究（7 - 28天及以后），丁丙诺啡（0.1-2.5毫克/公斤SQ）的剂量，将管理，每6小时手术后24-48小时的手术和卡洛芬口服片剂后，将每天提供一次（见“手术后恢复”）。 </li></ol> 2。大鼠的制备及插管<ol><li>动物麻醉诱导前，将审查，以确定任何预先存在的条件，可能会复杂手术的结果，体检将包括目视检查和测量心率，呼吸率，体温和体重。 </li><li>如果动物是健康的，将250-300克（例如只Dawley）雄性大鼠称重和识别特征将被记录。 </li><li>然后用80毫克/公斤苯巴比妥麻醉大鼠施加压力的大鼠甲床（趾捏反射），并定期整个过程，验证正确的麻醉水平是通过肢体的戒断反应。 </li><li>丁丙诺啡（0.1-2.5毫克/公斤; SQ）将管理在手术时间。 </li><li>然后用电动剃须刀刮胡子的皮毛形式的颈部和胸部区域。注：对于涉及过了一段时间的评估心脏功能和心脏重塑的研究（7 - 28天及以后），毛皮，必须完全剃光从手术区保持无菌。 </li><li>剃光地区擦洗消毒，并用70％酒精擦拭面积优碘的解决方案;此优碘和酒精擦洗和消毒程序是重复三次。 </li><li>麻醉动物，然后放置在一个homeothermic毯系统（电热垫）在仰卧位，以保持体温。 </li><li>然后进行气管插管，首先要仔细暴露气管，提高插管的可视化，通过一个5毫米的中期颈部切口和肌肉组织回缩，略高于气管。 </li><li>这使得油管上的优势轻松通过喉部的入口斜面，更好地插入气管内管（聚乙烯大小90，PE 90）的可视化。 </li><li>气管导管插入气管，使用手术立体声显微镜计算机辅助可视化，通过精心操纵的舌头，一边欣赏气管和气管导管的入口处。 </li><li>然后实现肯特科学的呼吸机（TOPO双模）在80和​​潮气量每100克体重1.2毫升每分钟呼吸循环连接气管插管机械通气。 </li><li>一个无菌的悬垂性，然后放置在大鼠，离开只有手术区暴露出来，从而最大限度地减少在操作过程中的潜在污染。 </li><li>动物外科医生和助手都戴上口罩，隔离衣，手套，手套之间的大鼠手术改变。 </li></ol> 3。瞬态左前降支（LAD）动脉结扎<ol><li>一旦建立稳定的呼吸，胸部打开左侧第四肋间，使用止血或回合结束剪刀，以开放的空间，但不切割，使出血的危险性可减少组织。 </li><li>胸部拉钩然后是定位在第四肋间;以传播的肋骨，使左心室（LV）的暴露，最大小心不要损坏肺。 </li><li>然后打开心包，但如果可能的话地方离开。 </li><li>将确定近端左前降支（LAD）动脉（图1），与使用作为urgical立体显微镜，法援署是那么短暂结扎（或可绑一个活结）用了30分钟的缺血时间（图1）6-0普理灵缝合，但没有外置的心。 </li><li>为了让心脏再灌注损伤，显微剪刀用于切割结6-0普理灵缝合，30分钟后在LAD结扎，结扎（或释放活结）。 </li><li>在深水对照组，程序是相同的，除法援署不是短暂结扎。 </li><li>流出然后简要（1-2秒）捏呼吸器关闭，以允许重新通胀的肺部​​。 </li><li>胸部拉钩，然后和排骨一起绘制，使用了2-0普理灵缝线间断缝合模式。 </li><li>一旦肋骨被关闭，流出呼吸再次简要地（1-2秒）捏关闭，以确保正确的呼吸。 </li><li>皮肤是关闭使用6-0普理灵缝线连续缝合模式。 </li></ol> 4。确认成功结扎的LAD动脉<ol><li>通过观察心肌组织的漂白以及功能障碍的前壁缝合远端正确结扎法援署将得到确认;在瞬态LAD结扎观察。 </li><li>再灌注红色返回到心肌组织和一些前壁运动恢复的示范，将验证;观察后立即结扎瞬态法援署。 </li><li>注：对于涉及过了一段时间的评估心脏功能和心脏重塑的研究（7 - 28天及以后），超声心动图将进行24小时后法援署结扎，以确定程度的诱导心肌梗死。左室短轴缩短率（LV FS）&lt;20％，超声心动图测定，以及前壁运动功能障碍与梗死的所有动物，将确定并用于后续心脏功能和重塑研究。深水对照组大鼠预计将有40-45％之间正常的LV FS（蒙特等）。 </li></ol> 5。手术后的恢复<ol><li>手术完成后，无菌悬垂在动物将被删除。要监测体温（直肠温度计），呼吸频率（目测），心率（心悸），异常体征和疼痛，直到动物卧床。 </li><li>老鼠将被删除，脱离呼吸机，然后放置在动物笼，并允许从麻醉中恢复。在这个笼子里，笼子的一端将升温加热垫，让大鼠寻求温暖。此外，水将可在此观察期间大鼠。 </li><li>如果脱水迹象，观察手术后，将注入0.5毫升无菌生理盐水腹腔注射（IP）。 </li><li>丁丙诺啡（0.1-2.5毫克/公斤; SQ）将被立即手术后，并在手术后的恢复管理。 </li><li>注：对于涉及过了一段时间的评估心脏功能和心脏重塑的研究（7 - 28天及以后），丁丙诺啡（0.1-2.5毫克/公斤SQ）的剂量，将管理，每6小时手术后24-48小时的手术后。卡洛芬（Rimadyl，Bioserv片剂）口服片剂也将提供至少48小时的手术后（1片剂每日）。 </li></ol> 6。缺血再灌注损伤的组织学评价<ol><li> 4小时再灌注期间完成后，老鼠是被麻醉后再次用80毫克/公斤苯巴比妥和法援署再次是结扎（或活结可以retied）（黄等，Kaiser等）。 </li><li> 1毫升5％埃文的蓝（磷酸盐缓冲液），然后注入右心室腔，并允许灌注2分钟。 </li><li> 2分钟后，1毫升饱和氯化钾溶液，然后注入右心室腔，以停止跳动的心。 </li><li>心是那么迅速切除，2％琼脂糖磷酸盐缓冲液中嵌入，然后在1-2毫米的片切片。 </li><li>存活心肌，然后2％氯化三苯四氮唑（TTC）染色20分钟，每个切片的图像被捕获。的图像，然后量化，以确定心肌面积金额：（1）是不是风险（蓝色部分），（2）区域风险（AAR，红色和白色部分），和（3 ）是心肌梗死的面积，（白色的“不染”的部分）（图2）。 </li></ol> 7。代表性的成果对于涉及梗死面积的评估和风险面积（AAR），埃文蓝色染料和氯化三苯四氮唑（TTC）的，下面的4小时再灌注的研究，妥善结扎法援署可确诊通过观察远端缝合以及功能障碍的前壁心肌组织的漂白，在瞬态LAD结扎观察。再灌注可以验证由红色返回到心肌组织和一些前壁运动恢复的示范;瞬态法援署结扎后立即观察。对于涉及过了一段时间的评估心脏功能和心脏重塑的研究（7 - 28 - 天及以后），缺血再灌注的正确结扎也可以得到证实，热烫的组织，功能障碍的前壁和室壁运动恢复与一些红色的回报，再灌注损伤。此外，心脏功能受损，可确定正在执行的24小时后法援署结扎超声心动图，以确定诱发心肌梗死的程度。左室短轴缩短率（LV FS）&lt;20％，超声心动图测定，以及前壁运动功能障碍与梗死的所有动物，将确定并用于后续心脏功能和重塑研究。深水对照组大鼠预计将有40-45％之间正常的LV FS（蒙特）。 <img src="/files/ftp_upload/2464/2464fig1.jpg" alt="图1" /> 图1。心肌梗死和LAD结扎的示意图。 （1）心肌梗死，左心室前降（LAD）动脉缝合，用来模拟由鼠疫形成引起的心肌梗死（2）结扎的示意图。 <img src="/files/ftp_upload/2464/2464fig2.jpg" alt="图2" /> 图2大鼠急性心肌梗死心脏的形象代表。大鼠呈现蓝色（非闭塞冠状动脉灌注区）区的中心地带的形象代表，在红（闭塞冠脉灌注，表明风险面积梗死（AAR）区面积，白/黄（梗死区面积在AAR内）。

<ol>
	<li>Hunt, S. A., Baker, D. W., Chin, M. H., Cinquegrani, M. P., Feldmanmd, A. M., Francis, G. S., Ganiats, T. G., Goldstein, S., Gregoratos, G., Jessup, M. L., Noble, R. J., Packer, M., Silver, M. A., Stevenson, L. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SC.ACC/AHA+Guidelines+for+the+Evaluation+and+Management+of+Chronic+Heart+Failure+in+the+Adult:+Executive+Summary+A+Report+of+the+American+College+of+Cardiology/American+Heart+Association+Task+Force+on+Practice+Guidelines+(Committee+to+Revise+the+1995+Guidelines+for+the+Evaluation+and+Management+of+Heart+Failure).">SC.ACC/AHA Guidelines for the Evaluation and Management of Chronic Heart Failure in the Adult: Executive Summary A Report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Committee to Revise the 1995 Guidelines for the Evaluation and Management of Heart Failure).</a> Circulation. 104, 2996-3007 (2001).</li><li>Goldman, S., Raya, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+infarct+model+of+myocardial+infarction+and+heart+failure.">Rat infarct model of myocardial infarction and heart failure.</a> J Card Fail. 1, 169-177 (1995).</li><li>Tarnavski, O., McMullen, J. R., Schinke, M., Nie, Q., Kong, S., &amp;amp, I. z. u. m. o., S, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+cardiac+surgery:+comprehensive+techniques+for+the+generation+of+mouse+models+of+human+diseases+and+their+application+for+genomic+studies.">Mouse cardiac surgery: comprehensive techniques for the generation of mouse models of human diseases and their application for genomic studies.</a> Physiol Genomics. 16, 349-360 (2004).</li><li>Thygesen, K., Alpert, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Universal+Definition+of+Myocardial+Infarction.">Universal Definition of Myocardial Infarction.</a> Circulation. 116, 2634-2653 (2007).</li><li>Vasilyev, N., Williams, T., Brennan, M. L., Unzek, S., Zhou, X., Heinecke, J. W., Spitz, D. R., Topol, E. J., Hazen, S. L., Penn, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myeloperoxidase-generated+oxidants+modulate+left+ventricular+remodeling+but+not+infarct+size+after+myocardial+infarction.">Myeloperoxidase-generated oxidants modulate left ventricular remodeling but not infarct size after myocardial infarction.</a> Circulation. 112, 2812-2820 (2005).</li><li>McConnell, B. K., Popovic, Z., Mal, N., Lee, K., Bautista, J., Forudi, F., Schwartzman, R., Jin, J. P., Penn, M., Bond, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+protein+kinase+A+interaction+with+A-kinase-anchoring+proteins+in+the+heart+in+vivo:+effects+on+cardiac+contractility,+protein+kinase+A+phosphorylation,+and+troponin+I+proteolysis.">Disruption of protein kinase A interaction with A-kinase-anchoring proteins in the heart in vivo: effects on cardiac contractility, protein kinase A phosphorylation, and troponin I proteolysis.</a> J Biol Chem. 284, 1583-1592 (2009).</li><li>Monte, D. e. l., Butler, F., Boecker, K., Gwathmey, W., K, J., Hajjar, R. 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C., Cheng, J., Perin, E., Fujise, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergistic+Infarct+Size+Reduction+by+Combination+Therapy+with+Milrinone+and+Esmolol+at+Early+Reperfusion+during+Acute+Myocardial+Infarction.">Synergistic Infarct Size Reduction by Combination Therapy with Milrinone and Esmolol at Early Reperfusion during Acute Myocardial Infarction.</a> American Journal of Physiology. , .</li><li>Kaiser, R. A., Bueno, O. F., Lips, D. J., Doevendans, P. A., Jones, F., Kimball, T. F., Molkentin, J. 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大鼠急性心肌梗死（AMI），也称为缺血再灌注（IR），是常用来模仿人类的​​心室重构，导致心力衰竭。在动物模型（小鼠和大鼠）的左室功能不全和心力衰竭的进展是维持心肌梗死，生存和随后发展心力衰竭（高盛）在人类观察到的类似。此过程概述了这项技术的一个版本，还载有附加说明描述方法学的变化。急性心肌梗死（AMI）过程的更多的这种变化，包括缺血时间和再灌注的时间。另外?...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Ventilator</td>
<td>&nbsp;</td>
<td>Kent Scientific</td>
<td>TOPO</td>
<td>TOPO Dual Mode</td>
</tr>
<tr>
<td>Thermostatic Circulator E103</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>732802</td>
<td>&nbsp;</td>
<tr>
<td>Homeothermic Blanket System</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>55-7222F</td>
<td>&nbsp;</td>
<tr>
<td>Stereo Microscope</td>
<td>&nbsp;</td>
<td>Nikon Instruments</td>
<td>MMA36300</td>
<td>SMZ-645 Zoom Stereo Microscope</td>
</tr>
<tr>
<td>Hot Bead Sterilizer</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>610183</td>
<td>&nbsp;</td>
<tr>
<td>Electric Shaver</td>
<td>&nbsp;</td>
<td>General Electric</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Blades and Handles</td>
<td>&nbsp;</td>
<td>WelchAllyn</td>
<td>690047-1</td>
<td>Standard (Lamp) Laryngoscope Blades</td>
</tr>
<tr>
<td>Surgical Scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14014-17</td>
<td>&nbsp;</td>
<tr>
<td>Surgical Scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14072-10</td>
<td>&nbsp;</td>
<tr>
<td>Forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11028-15</td>
<td>&nbsp;</td>
<tr>
<td>Curved Forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11001-12</td>
<td>&nbsp;</td>
<tr>
<td>Needle Holder</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>12004-18</td>
<td>&nbsp;</td>
<tr>
<td>Needle Holder</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>91201-13</td>
<td>&nbsp;</td>
<tr>
<td>Chest Retractor</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>17008-07</td>
<td>&nbsp;</td>
<tr>
<td>Masks</td>
<td>&nbsp;</td>
<td>VWR </td>
<td>47730-640</td>
<td>&nbsp;</td>
<tr>
<td>Gauze Sponges</td>
<td>&nbsp;</td>
<td>Dermacea</td>
<td>441205</td>
<td>&nbsp;</td>
<tr>
<td>Sterile Gloves</td>
<td>&nbsp;</td>
<td>Cardinal Health</td>
<td>2D7203I</td>
<td>&nbsp;</td>
<tr>
<td>PE 90 tubing</td>
<td>&nbsp;</td>
<td>BD</td>
<td>427420</td>
<td>&nbsp;</td>
<tr>
<td>20 ml Syringe</td>
<td>&nbsp;</td>
<td>BD</td>
<td>301625</td>
<td>&nbsp;</td>
<tr>
<td>6-0 Prolene Suture</td>
<td>&nbsp;</td>
<td>ETHICON</td>
<td>8706H</td>
<td>&nbsp;</td>
<tr>
<td>2-0 Silk Suture</td>
<td>&nbsp;</td>
<td>ETHICON</td>
<td>K873H</td>
<td>&nbsp;</td>
<tr>
<td>Pentobarbital Sodium Injection</td>
<td>&nbsp;</td>
<td>OVATION</td>
<td>NDC 67386-501-55</td>
<td>&nbsp;</td>
<tr>
<td>Povidone-Iodine</td>
<td>&nbsp;</td>
<td>NOVAPLUS</td>
<td>V10-8204</td>
<td>&nbsp;</td>
<tr>
<td>Buprenorphine</td>
<td>&nbsp;</td>
<td>Pharmaceuticals Inc.</td>
<td>NDC 12496-0757-1</td>
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		</div>

acute myocardial infarction in rats

（亨特）在美国领导的死因与心脏衰竭，生物医学研究的基本推动心血管疾病的药物治疗。动物模型，模仿人类的​​心脏疾病，如心肌梗死（MI）和缺血再灌注（IR），导致心力衰竭以及压力超负荷（横向主动脉缩窄），引起心肌肥厚和心力衰竭（高盛和Tarnavski）是有用的模型来研究心血管疾病。特别是，心肌缺血（MI）是一个导致尽管控制一定的风险因素，如动脉硬化，并通过手术干预（Thygesen）治疗心血管疾病的发病率和死亡率。此外，心肌缺血（MI）的结果在急性心肌损失，增加负载条件下，诱导心室重塑和远程非梗死心肌梗死的边境地带。心肌细胞凋亡，坏死，并由此增加的血流动力学负载激活多种生化细胞内的信号，启动LV扩张，肥厚，心室的形状失真，和胶原瘢痕形成。这种病理重塑和未能正常化，增加墙强调，在逐步扩张，招聘的边境地区到瘢痕心肌，并最终在心肌收缩功能的恶化（即心力衰竭）的结果。维持一个大的心肌梗死，生存和随后的发展心力衰竭（高盛）的病人中观察到类似的左室功能不全和心力衰竭大鼠的进展。急性心肌梗死（AMI）大鼠模型一直被用来模仿人类的​​心血管疾病;专门用于研究心脏信号与心力衰竭，以及评估治疗心力衰竭的治疗策略的贡献相关的机制。在这份报告中所描述的方法是急性心肌梗死（AMI）大鼠模型。这种模式也称为其次再灌注（IR）;这是一种急性结扎左前降支（LAD）30分钟的时间内组织再灌注缺血诱导的急性缺血性心肌病或缺血释放的LAD结扎（瓦西里耶夫和McConnell）。此协议将重点和的面积，在，风险评估梗死面积（AAR）埃文的蓝色染料和三苯基四氮唑氯化四氮唑（TTC）再灌注4 -小时;走向评价心脏功能和重塑额外的意见修改再灌注期间，还赠送。总体而言，这一急性心肌梗死大鼠动物模型，研究了对心脏的病理生理和生理功能的心肌梗死的后果非常有用。

Watch this Scientific Journal Video about Acute Myocardial Infarction in Rats at JoVE.com

Acute Myocardial Infarction in Rats

急性心肌梗死（AMI）大鼠模型是有用的研究对心脏的病理生理及生理功能的心肌梗死的后果。

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 ...

Research

JoVE Journal

Medicine

52.6K 次观看.  University of Texas Medical Branch. 急性心肌梗死（AMI）大鼠模型是有用的研究对心脏的病理生理及生理功能的心肌梗死的后果。

视频: Acute Myocardial Infarction in Rats

Induction of Myocardial Infarction in Adult Zebrafish Using Cryoinjury

The mammalian heart is incapable of significant regeneration following an acute injury such as myocardial infarction1. By contrast, urodele amphibians and teleost fish retain a remarkable capacity for cardiac regeneration with little or no scarring throughout life2,3. It is not known why only some non-mammalian vertebrates can recreate a complete organ from remnant tissues4,5. To understand the molecular and cellular differences between regenerative responses in different species, we need to use similar approaches for inducing acute injuries.
In mammals, the most frequently used model to study cardiac repair has been acute ischemia after a ligation of the coronary artery or tissue destruction after cryoinjury6,7. The cardiac regeneration in newts and zebrafish has been predominantly studied after a partial resection of the ventricular apex2,3. Recently, several groups have established the cryoinjury technique in adult zebrafish8-10. This method has a great potential because it allows a comparative discussion of the results obtained from the mammalian and non-mammalian species.
Here, we present a method to induce a reproducible disc-shaped infarct of the zebrafish ventricle by cryoinjury. This injury model is based on rapid freezing-thawing tissue, which results in massive cell death of about 20% of cardiomyocytes of the ventricular wall. First, a small incision was made through the chest with iridectomy scissors to access the heart. The ventricular wall was directly frozen by applying for 23-25 seconds a stainless steel cryoprobe precooled in liquid nitrogen. To stop the freezing of the heart, fish water at room temperature was dropped on the tip of the cryoprobe. The procedure is well tolerated by animals, with a survival rate of 95%.
To characterize the regenerative process, the hearts were collected and fixed at different days after cryoinjury. Subsequently, the specimen were embedded for cryosectioning. The slides with sections were processed for histological analysis, in situ hybridization and immunofluorescence. This undertaking enhances our understanding of the factors that are required for the regenerative plasticity in the zebrafish, and provide new insights into the machinery of cardiac regeneration. A conceptual and molecular understanding of heart regeneration in zebrafish will impact both developmental biology and regenerative medicine.

Zebrafish represents a valuable model to study the mechanisms of heart regeneration in vertebrates. Here, we present a protocol for induction of a heart infarct in adult zebrafish using cryoinjury. This method results in massive cell death within 20% of the ventricular wall, similar to that observed in mammalian infarcts.

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 ...

科研

医学

MRI and PET in Mouse Models of Myocardial Infarction

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 ideal model for testing novel therapeutic strategies.
In vivo magnetic resonance imaging (MRI) gives excellent views of the heart noninvasively with clear anatomical detail, which can be used for accurate functional assessment. Contrast agents can provide basic measures of tissue viability but these are nonspecific. Positron emission tomography (PET) is a complementary technique that is highly specific for molecular imaging, but lacks the anatomical detail of MRI. Used together, these techniques offer a sensitive, specific and quantitative tool for the assessment of the heart in disease and recovery following treatment.
In this paper we explain how these methods are carried out in mouse models of acute myocardial infarction. The procedures described here were designed for the assessment of putative protective drug treatments. We used MRI to measure systolic function and infarct size with late gadolinium enhancement, and PET with fluorodeoxyglucose (FDG) to assess metabolic function in the infarcted region. The paper focuses on practical aspects such as slice planning, accurate gating, drug delivery, segmentation of images, and multimodal coregistration. The methods presented here achieve good repeatability and accuracy maintaining a high throughput.

We describe how to perform MRI and PET imaging of the mouse heart. The protocol is tailored to assess treatment efficacy in models of myocardial infarction and heart failure.

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

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One important prerequisite is a good understanding of all safety and efficacy aspects obtained in a large animal model that validly reflect the human scenario of myocardial infarction (MI). Pigs are widely used in this regard since their cardiac size, hemodynamics, and coronary anatomy are close to that of humans. Here, we present an effective protocol for using the porcine MI model using a closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. This approach is based on 90 min of myocardial ischemia, inducing large left ventricle infarction of the anterior, septal and inferoseptal walls. Furthermore, we present protocols for various measures of outcome that provide a wide range of information on the heart, such as cardiac systolic and diastolic function, hemodynamics, coronary flow velocity, microvascular resistance, and infarct size. This protocol can be easily tailored to meet study specific requirements for the validation of novel cardioregenerative biologics at different stages (i.e. directly after the acute ischemic insult, in the subacute setting or even in the chronic MI once scar formation has been completed). This model therefore provides a useful translational tool to study MI, subsequent adverse remodeling, and the potential of novel cardioregenerative agents.

This protocol describes the porcine myocardial infarction (MI) model using a 90 min closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. Furthermore, the protocol for several outcome parameters, such as cardiac function, hemodynamics, microvascular resistance, and infarct size, are also presented.

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 ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last decades. For clarification of the underlying mechanisms and the development of new therapeutic strategies, the availability of valid animal models are mandatory. Since we need new insights into pathomechanisms of cardiovascular diseases under in vivo conditions to combat myocardial infarction, the validity of the animal model is a crucial aspect. However, protection of animals are highly relevant in this context. Therefore, we establish a minimally invasive and simple model of myocardial infarction in mice, which assures a high reproducibility and survival rate of animals. Thus, this models fulfils the requirements of the 3R principle (Replacement, Refinement and Reduction) for animal experiments and assure the scientific information needed for further developing of therapeutical strategies for cardiovascular diseases.

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

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 ...

Primary Outcome Assessment in a Pig Model of Acute Myocardial Infarction

Mortality after acute myocardial infarction remains substantial and is associated with significant morbidity, like heart failure. Novel therapeutics are therefore required to confine cardiac damage, promote survival and reduce the disease burden of heart failure. Large animal experiments are an essential part in the translational process from experimental to clinical therapies. To optimize clinical translation, robust and representative outcome measures are mandatory. The present manuscript aims to address this need by describing the assessment of three clinically relevant outcome modalities in a pig acute myocardial infarction (AMI) model: infarct size in relation to area at risk (IS/AAR) staining, 3-dimensional transesophageal echocardiography (TEE) and admittance-based pressure-volume (PV) loops. Infarct size is the main determinant driving the transition from AMI to heart failure and can be quantified by IS/AAR staining. Echocardiography is a reliable and robust tool in the assessment of global and regional cardiac function in clinical cardiology. Here, a method for three-dimensional transesophageal echocardiography (3D-TEE) in pigs is provided. Extensive insight into cardiac performance can be obtained by admittance-based pressure-volume (PV) loops, including intrinsic parameters of myocardial function that are pre- and afterload independent. Combined with a clinically feasible experimental study protocol, these outcome measures provide researchers with essential information to determine whether novel therapeutic strategies could yield promising targets for future testing in clinical studies.

Reliable and accurate outcome assessment is the key for translation of preclinical therapies into clinical treatment. The current paper describes how to assess three clinically relevant primary outcome parameters of cardiac performance and damage in a pig acute myocardial infarction model.

急性心肌梗死的猪模型主要成果评估

Mortality after acute myocardial infarction remains substantial and is associated with significant morbidity, like heart failure. Novel therapeutics are ...

Myocardial Infarction in Neonatal Mice, A Model of Cardiac Regeneration

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 regeneration, and to define new targets for therapeutics. For decades, models of complete heart regeneration existed in amphibians and fish, but a mammalian counterpart was not available. The recent discovery of a postnatal window during which mice possess regenerative capabilities has led to the establishment of a mammalian model of cardiac regeneration. A surgical model of mammalian cardiac regeneration in the neonatal mouse is presented herein. Briefly, postnatal day 1 (P1) mice are anesthetized by isoflurane and placed on an ice pad to induce hypothermia. After the chest is opened, and the left anterior descending coronary artery (LAD) is visualized, a suture is placed around the LAD to inflict myocardial ischemia in the left ventricle. The surgical procedure takes 10-15 min. Visualizing the coronary artery is crucial for accurate suture placement and reproducibility. Myocardial infarction and cardiac dysfunction are confirmed by triphenyl-tetrazolium chloride (TTC) staining and echocardiography, respectively. Complete regeneration 21 days post myocardial infarction is verified by histology. This protocol can be used to as a tool to elucidate mechanisms of mammalian cardiac regeneration after myocardial infarction.

This protocol describes a highly reproducible model of cardiac regeneration by surgical induction of myocardial infarction in the left ventricle of postnatal day 1 mice. The method involves induction of hypothermic anesthesia and ligation of the left anterior descending coronary artery.

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 ...

Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any potential therapeutic approach directed against acute left ventricular (LV) dysfunction. Current imaging modalities (e.g., echocardiography and magnetic resonance imaging) have several limitations since data on LV pressure cannot directly be measured. LV catheterization in mice undergoing coronary artery occlusion could serve as a novel method for a real-time evaluation of LV function.
At the beginning of the procedure, mice were anesthetized followed by endotracheal intubation. For LV catheterization, the right carotid artery was exposed via middle-neck incision. The catheter was introduced and placed into the LV cavity. Left thoracotomy was conducted and the left main coronary artery (LCA) was ligated. To induce reperfusion, the suture was released after 45 min. Pressure-volume data was recorded at all times.
Ligation of the LCA caused a decrease in LV systolic function as evidenced by a 30% reduction in stroke volume, LV ejection fraction (EF), and cardiac output. Maximum dP/dt as a parameter for LV contractility was also significantly reduced and diastolic function was severely impaired (minimum dP/dt -40%). Reperfusion over a period of 20 min did not lead to a complete recovery of LV function.
Real-time pressure-volume analysis served as a valid procedure for monitoring cardiac function during acute myocardial infarction in mice. Maintaining stable anesthesia and a standardized surgical approach was crucial to ensure valid results. As the early phase of acute myocardial infarction is critical for morbidity and mortality, the delineated method could be beneficial for preclinical evaluation of new strategies for cardioprotection.

Acute myocardial infarction in mice induces acute but incompletely characterized changes in left ventricular (LV) function. LV catheterization in mice undergoing coronary artery occlusion serves as a novel method for a real-time evaluation of LV function.

小鼠急性心肌梗死的实时压力容积分析

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any ...

Digital PCR for Quantifying Circulating MicroRNAs in Acute Myocardial Infarction and Cardiovascular Disease

Circulating serum microRNAs (miRNAs) have shown promise as biomarkers for the cardiovascular disease and acute myocardial infarction (AMI), being released from the cardiovascular cells into the circulation. Circulating miRNAs are highly stable and can be quantified. The quantitative expression of specific miRNAs can be linked to the pathology, and some miRNAs show high tissue and disease specificity. Finding novel biomarkers for cardiovascular diseases is of importance for medical research. Quite recently, digital polymerase chain reaction (dPCR) has been invented. dPCR, combined with fluorescent hydrolysis probes, enables specific direct absolute quantification. dPCR exhibits superior technical qualities, including a low variability, high linearity, and high sensitivity compared to the quantitative polymerase chain reaction (qPCR). Thus, dPCR is a more accurate and reproducible method for directly quantifying miRNAs, particularly for the use in large multi-center cardiovascular clinical trials. In this publication, we describe how to effectively perform digital PCR in order to assess the absolute copy number in serum samples.

Circulating microRNAs have shown promise as biomarkers for cardiovascular diseases and acute myocardial infarctions. In this study, we describe a protocol for miRNA extraction, reverse transcription, and digital PCR for the absolute quantification of miRNAs in the serum of patients with cardiovascular disease.

数字 PCR 在急性心肌梗死和心血管疾病中量化循环 MicroRNAs 的临床观察

Circulating serum microRNAs (miRNAs) have shown promise as biomarkers for the cardiovascular disease and acute myocardial infarction (AMI), being released ...

A Cryoinjury Model to Study Myocardial Infarction in the Mouse

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we demonstrate a murine cryoinjury infarct model that generates precise infarct sizes with high reproducibility and replicability. In brief, after intubation and sternotomy of the animal, the heart is lifted from the thorax. The probe of a handheld liquid nitrogen delivery system is applied onto the myocardial wall to induce cryoinjury. Impaired ventricular function and electrical conduction can be monitored with echocardiography or optical mapping. Transmural myocardial remodeling of the infarcted area is characterized by collagen deposition and loss of cardiomyocytes. Compared to other models (e.g., LAD-ligation), this model utilizes a handheld liquid nitrogen delivery system to generate more uniform infarct sizes.

This article demonstrates a model to study cardiac remodeling after myocardial cryoinjury in mice.

研究小鼠心肌梗死的低温损伤模型

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we ...

Semi-Minimal Invasive Method to Induce Myocardial Infarction in Rats and the Assessment of Cardiac Function by an Isolated Working Heart System

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily and highly reproducible MI induction procedure is required to obtain further insight and better understanding of the underlying pathological changes. This procedure can also be used to evaluate the effects or potency of new and promising treatments (as drugs or interventions) in acute MI, subsequent remodeling and heart failure (HF). After intubation and pre-operative preparation of the animal, an anesthetic protocol with isoflurane was performed, and the surgical procedure was conducted&#160;quickly. Using a minimally invasive approach, the left anterior descending artery (LAD) was located and occluded by a ligature. The occlusion can be performed acutely for subsequent reperfusion (ischemia/reperfusion injury). Alternatively, the vessel can be ligated permanently to investigate the development of chronic MI, remodeling or HF. Despite common pitfalls, the drop-out rates are minimal. Various treatments such as remote ischemic conditioning can be examined for their cardioprotective potential pre-, peri- and post-operatively. The post-operative recovery was quick as the anesthesia was precisely controlled and the duration of the operation was short. Post-operative analgesia was administered for three days. The minimally invasive procedure reduces the risk of infection and inflammation. Furthermore, it facilitates rapid recovery. The &#8220;working heart&#8221; measurements were performed ex vivo and enabled precise control of preload, afterload and flow. This procedure requires specific equipment and training for adequate performance. This manuscript provides a detailed step-by-step introduction for conducting these measurements.

This article presents an efficient method to perform myocardial ischemia and subsequent chronic reperfusion in rats using a minimally invasive approach. In addition, left ventricular hemodynamic function of rats is assessed by echocardiography and isolated working heart methods.&#160;

诱导大鼠心肌梗死的半微创方法及独立工作心脏系统对心脏功能的评估

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily ...