Name: implantation of an isoproterenol mini-pump to induce heart failure in mice
Uploaded: 2019-10-03T13:00:00
Description: Watch this Scientific Journal Video about Implantation of an Isoproterenol Mini-Pump to Induce Heart Failure in Mice at JoVE.com

作者承认NIH K08 HL133491的资金支持。

该协议符合加州大学洛杉矶分校的动物护理指南（ARC 协议#2010-075）。建议读者遵守自己的IACUC批准的协议，因为近程序小鼠护理和麻醉管理可能是特定于机构的。
1. 制备异丙酮醇渗透泵
注：此程序已成功应用于9周以上雌性小鼠，体重超过18克，来自100多个近亲繁殖小鼠菌株，以及一个菌株子集的雄性小鼠。此过程没有最大重量限制。始终包括年龄匹配的?...

<ol>
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P., Singh, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+of+heart+failure.">Rodent models of heart failure.</a> Journal of Pharmacological and Toxicological Methods. 56 (1), 1-10 (2007).</li><li>Patten, R. D., Hall-Porter, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+animal+models+of+heart+failure:+development+of+novel+therapies,+past+and+present.">Small animal models of heart failure: development of novel therapies, past and present.</a> Circulation: Heart Failure. 2 (2), 138-144 (2009).</li><li>Huang, W. Y., Aramburu, J., Douglas, P. S., Izumo, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+expression+of+green+fluorescence+protein+can+cause+dilated+cardiomyopathy.">Transgenic expression of green fluorescence protein can cause dilated cardiomyopathy.</a> Nature Medicine. 6 (5), 482-483 (2000).</li><li>Breckenridge, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+failure+and+mouse+models.">Heart failure and mouse models.</a> Disease Model and Mechanism. 3 (3-4), 138-143 (2010).</li><li>Wittstein, I. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurohumoral+features+of+myocardial+stunning+due+to+sudden+emotional+stress.">Neurohumoral features of myocardial stunning due to sudden emotional stress.</a> New England Journal of Medicine. 352 (6), 539-548 (2005).</li><li>Templin, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+Features+and+Outcomes+of+Takotsubo+(Stress)+Cardiomyopathy.">Clinical Features and Outcomes of Takotsubo (Stress) Cardiomyopathy.</a> New England Journal of Medicine. 373 (10), 929-938 (2015).</li><li>Shao, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mouse+model+reveals+an+important+role+for+catecholamine-induced+lipotoxicity+in+the+pathogenesis+of+stress-induced+cardiomyopathy.">A mouse model reveals an important role for catecholamine-induced lipotoxicity in the pathogenesis of stress-induced cardiomyopathy.</a> European Journal of Heart Failure. 15 (1), 9-22 (2013).</li><li>Kudej, R. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+chronic+beta-adrenergic+receptor+stimulation+in+mice.">Effects of chronic beta-adrenergic receptor stimulation in mice.</a> Journal of Molecular Cell Cardiology. 29 (10), 2735-2746 (1997).</li><li>Zhuo, X. Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoproterenol+instigates+cardiomyocyte+apoptosis+and+heart+failure+via+AMPK+inactivation-mediated+endoplasmic+reticulum+stress.">Isoproterenol instigates cardiomyocyte apoptosis and heart failure via AMPK inactivation-mediated endoplasmic reticulum stress.</a> Apoptosis. 18 (7), 800-810 (2013).</li><li>El-Demerdash, E., Awad, A. S., Taha, R. M., El-Hady, A. M., Sayed-Ahmed, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Probucol+attenuates+oxidative+stress+and+energy+decline+in+isoproterenol-induced+heart+failure+in+rat.">Probucol attenuates oxidative stress and energy decline in isoproterenol-induced heart failure in rat.</a> Pharmacology Research. 51 (4), 311-318 (2005).</li><li>Rau, C. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+genetic+contributions+to+cardiac+pathology+induced+by+Beta-adrenergic+stimulation+in+mice.">Mapping genetic contributions to cardiac pathology induced by Beta-adrenergic stimulation in mice.</a> Circulation Cardiovascular Genetics. 8 (1), 40-49 (2015).</li><li>Berthonneche, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+response+to+beta-adrenergic+blockade+or+activation+in+23+inbred+mouse+strains.">Cardiovascular response to beta-adrenergic blockade or activation in 23 inbred mouse strains.</a> PLoS One. 4 (8), 6610 (2009).</li><li>Chang, S. C., Ren, S., Rau, C. D., Wang, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoproterenol-Induced+Heart+Failure+Mouse+Model+Using+Osmotic+Pump+Implantation.">Isoproterenol-Induced Heart Failure Mouse Model Using Osmotic Pump Implantation.</a> Methods Molecular Biology. 1816, 207-220 (2018).</li><li>Shusterman, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strain-specific+patterns+of+autonomic+nervous+system+activity+and+heart+failure+susceptibility+in+mice.">Strain-specific patterns of autonomic nervous system activity and heart failure susceptibility in mice.</a> American Journal of Physiology-Heart Circulation Physiology. 282 (6), 2076-2083 (2002).</li><li>Faulx, M. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strain-dependent+beta-adrenergic+receptor+function+influences+myocardial+responses+to+isoproterenol+stimulation+in+mice.">Strain-dependent beta-adrenergic receptor function influences myocardial responses to isoproterenol stimulation in mice.</a> American Journal of Physiology-Heart Circulation Physiology. 289 (1), 30-36 (2005).</li><li>Zhang, G. X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+oxidative+stress+in+acute+and+chronic+isoproterenol-infused+rats.">Cardiac oxidative stress in acute and chronic isoproterenol-infused rats.</a> Cardiovascular Research. 65 (1), 230-238 (2005).</li><li>Boluyt, M. O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoproterenol+infusion+induces+alterations+in+expression+of+hypertrophy-associated+genes+in+rat+heart.">Isoproterenol infusion induces alterations in expression of hypertrophy-associated genes in rat heart.</a> American Journal of Physiology. 269 (2), 638-647 (1995).</li><li>Hohimer, A. R., Davis, L. E., Hatton, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repeated+daily+injections+and+osmotic+pump+infusion+of+isoproterenol+cause+similar+increases+in+cardiac+mass+but+have+different+effects+on+blood+pressure.">Repeated daily injections and osmotic pump infusion of isoproterenol cause similar increases in cardiac mass but have different effects on blood pressure.</a> Canadian Journal of Physiology and Pharmacology. 83 (2), 191-197 (2005).</li><li>Cross, H. R., Murphy, E., Koch, W. J., Steenbergen, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Male+and+female+mice+overexpressing+the+beta(2)-adrenergic+receptor+exhibit+differences+in+ischemia/reperfusion+injury:+role+of+nitric+oxide.">Male and female mice overexpressing the beta(2)-adrenergic receptor exhibit differences in ischemia/reperfusion injury: role of nitric oxide.</a> Cardiovascular Research. 53 (2), 662-671 (2002).</li><li>Klingman, G. I., McKay, G., Ward, A., Morse, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+isoproterenol+treatment+of+mice:+effects+on+catecholamines+and+rectal+temperature.">Chronic isoproterenol treatment of mice: effects on catecholamines and rectal temperature.</a> Journal of Pharmaceutical Sciences. 62 (5), 798-801 (1973).</li></ol>

我们已经将这种方法应用于超过100多种近亲小鼠，以评估慢性β-肾上腺素刺激引起的心脏结果2，13。已知在小鼠菌株中存在异丙酮醇的易感性显著差异，可根据所需兴趣菌株进行定制16。这可能是由于小鼠菌株17之间β-肾上腺素受体功能的变化，自主神经系统活性16的菌株依赖性差异，β-肾上腺素受?...

心力衰竭与减少弹出分数（HFrEF）伴随着一个公认的补偿反应的同情神经系统，以保持心血管平衡1。慢性活化观察到血液动力学压力和对心脏和循环的有害影响。这些已成为当代心力衰竭药物治疗的基石，是心力衰竭和神经激素系统治疗对抗发展的重要机制1。
有几个小鼠模型可用于心脏衰竭的基本调查。基因模型对探索分子疗法和研究信号通路很有吸引力。然而，这些模型可能与常见的心力衰竭形式无关。其他常见模型包括左前下降 （LAD） 动脉结扎、 横端收缩 （TAC） 和异丙酮醇 （ISO），每个模型针对不同的病理病因2、3、4、5 ，6.LAD动脉结扎诱导前壁心肌梗死，从而创建缺血性心肌病的专用模型。TAC诱导急性压力过载，形成心力衰竭的高血压模型。虽然可以测量压力梯度，允许肥大的分层，但高血压的急性发病缺乏直接的临床相关性4。LAD 和 TAC 型号都需要高水平的外科专业知识才能执行。急性ISO心力衰竭模型模仿应激引起的心肌病，也称为Takotsubo病，其特征是卡他索明的明显增加和左心室的活性，模仿急性心肌梗死7， 8.相比之下，慢性ISO型心力衰竭模型存在晚期心力衰竭的症状特征，慢性高浓度的儿茶酚胺1。慢性 ISO 模型的优点是，它提供模仿晚期心力衰竭的慢性肾上腺素刺激，并且相对容易产生。研究者应该选择一个模型，最好地概括他们感兴趣的病理学。
这种方法的总体目标是诱导小鼠的心力衰竭，使用植入的微型泵，释放 ISO 持续模仿慢感活化发现心力衰竭患者 1 。该方法简单易用。虽然小鼠菌株之间有明显差异，但以30毫克/千克/天在30mg/kg/天下施用3至4周的ISO足以诱导大多数小鼠进行心脏重塑。具体来说，ISO在第1周导致亲肥大补偿阶段，随后在第2周和第32周出现壁变薄、心室扩张和收缩功能下降。ISO模型创建的成功可以通过连续的超声心动图在体内评估，证明肥大、扩张和心室功能障碍，以及通过组织学和分子评估采集的心脏组织进行肌萎缩性功能评估。脂质积累，纤维化，ER压力，凋亡，和基因表达9，10，11，12。

<table>
	<tbody>
		<tr>
			<td>Micro-Osmotic Pump System with Flow Moderator in Place</td>
			<td>Alzet</td>
			<td>Model 1004</td>
			<td>Includes filling tube, flow moderator and pump body</td>
		</tr>
		<tr>
			<td>(-)-Isoproterenol hydrochloride</td>
			<td>Sigma-Aldrich</td>
			<td>16504-1G</td>
			<td>(-)-Isoproterenol hydrochloride is a powder that needs to be stored at -20&#176;C.</td>
		</tr>
		<tr>
			<td>1 ml sterile syringe</td>
			<td>VWR</td>
			<td>BD309602</td>
			<td></td>
		</tr>
		<tr>
			<td>30 W LED Fiber optic O-ring light microscope illuminator</td>
			<td>AmScope</td>
			<td>SKU: LED-30WR</td>
			<td></td>
		</tr>
		<tr>
			<td>5-0 COATED VICRYL (polyglactin 910) Suture</td>
			<td>Ethicon</td>
			<td>J303H</td>
			<td>5-0, absorbable</td>
		</tr>
		<tr>
			<td>Fine Scissors - Sharp</td>
			<td>FST</td>
			<td>14060-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass beads</td>
			<td>FST</td>
			<td>18000-46</td>
			<td></td>
		</tr>
		<tr>
			<td>Hot bead sterilizers</td>
			<td>FST</td>
			<td>18000-50</td>
			<td></td>
		</tr>
		<tr>
			<td>Iris forceps</td>
			<td>WPI</td>
			<td>15915</td>
			<td></td>
		</tr>
		<tr>
			<td>Look Sharpoint 6-0, 18&#34; Black Nylon Monofilament Suture</td>
			<td>LOOK</td>
			<td>AA-2176</td>
			<td>6-0, non-absorbable</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>WPI</td>
			<td>15926</td>
			<td></td>
		</tr>
		<tr>
			<td>Normal Saline, 0.9% NaCl</td>
			<td>Fisher</td>
			<td>89167-772</td>
			<td></td>
		</tr>
	</tbody>
</table>

implantation of an isoproterenol mini-pump to induce heart failure in mice

Isoproterenol（ISO）是一种非选择性β-肾上腺素激动剂，广泛用于诱导小鼠心脏损伤。虽然急性模型模仿压力引起的心肌病，而通过渗透泵施用的慢性模型模仿人类晚期心力衰竭。所述协议的目的是使用植入的微型泵在小鼠中创建慢性ISO引起的心力衰竭模型。该协议已用于诱导100+系近亲繁殖小鼠的心力衰竭。手术泵植入技术被详细描述，可能与任何有兴趣在小鼠中创建心力衰竭模型的人相关。此外，还介绍了基于每个菌株的超声心动图参数和模型开发的预期时间，每周心脏重塑变化。总之，该方法简单易用。通过植入式微型泵在3至4周内连续施用ISO，足以诱导心脏重塑。最后，通过连续超声心动图显示肥大、心室扩张和功能障碍，可以评估ISO模型创建的成功。

在我们之前发表的研究中，我们通过渗透泵在105种混合小鼠多样性面板（HMDP）菌株2、13中施用ISO剂量为30mg/kg/d。我们使用在ISO治疗的基线第1、2和3周进行的超声心动图评估结果（图1）。与先前的研究类似，作者研究了异丙酮醇在23个小鼠菌株14中的影响，我们观察到超声心动图参数之间惊人的菌株间变异。根据?...

Watch this Scientific Journal Video about Implantation of an Isoproterenol Mini-Pump to Induce Heart Failure in Mice at JoVE.com

Implantation of an Isoproterenol Mini-Pump to Induce Heart Failure in Mice

通过植入的渗透泵长期给用异丙酮醇，已被广泛用于模拟小鼠的晚期心力衰竭。在这里，我们描述了手术微型泵植入在持续异丙酮醇给3周内的详细方法，以及成功创建模型的超声心动图评估。

植入异丙酮醇微型泵，以诱导小鼠心力衰竭

Isoproterenol (ISO), is a non-selective beta-adrenergic agonist, that is used widely to induce cardiac injury in mice. While the acute model mimics ...

通过渗透泵长期给给异丙醇已被广泛用于综述小鼠晚期心力衰竭的变化。这种方法很简单，需要比其他模型，如LAD和TAC更少的手术专业知识。王一斌博士实验室的研究科学家任文森博士将证明这一程序。在称量和记录每只小鼠的体重后，计算每种动物的适当量和异丙醇浓度，并在120微升无菌0.9%氯化钠中溶解异丙醇。彻底混合一分钟，完全溶解异丙醇，并称重并记录一个空渗透泵的重量，以及和没有其流量调节器，每只鼠标。连接一个 27 规格的钝尖灌装管，随渗透泵一起安装一个 1 毫升注射器，每只鼠标有 120 微升的异丙醇溶液。将泵保持直立位置，将灌装管插入顶部的泵开口，直到灌装管的尖端靠近泵储液罐的底部，然后缓慢地按下注射器柱塞，直到异丙醇溶液填充到泵开口。装满泵后，小心地拆下灌装管，并擦去多余的溶液。插入回流量调节器以关闭泵，并擦去多余的溶液。然后重新加压泵，确认超过 90% 的储液罐容量已加注。将鼠标放在加热垫上的上部位置，然后从下腹部去除空气。对于手术性渗透泵植入，确认麻醉小鼠对手趾捏缺乏反应，并在动物的眼睛上涂抹软膏。用合适的消毒剂清洁裸露的皮肤，并制作一厘米的中线皮肤切口。使用一把钝端剪刀仔细解剖下皮面壁的皮肤，并使用钳子将内皮壁从底层肠中拉出。使用细剪刀在腹膜壁上切割一个0.8厘米的孔，然后首先将渗透泵流量调节器端插入腹腔。当泵就位时，使用 5-0 可吸收的缝合线以中断的方式关闭内面壁上的孔，并使用 6-0 不可吸收的缝合线关闭皮肤。然后将鼠标放在一个专门的孵化器中，进行监控，直到完全保持，然后再将动物返回到其家庭笼子。此处，介绍了基于所选超声心动图参数的每周变化摘要以及建模的预期时间。虽然观察到小鼠菌株之间的变化，平均而言，在结束硅藻和分数缩短增加在第一周，但减少以后的时间点。左心室内径在端膜和左心室质量增加在三个星期内。此外，小鼠心脏在异丙醇下显示明显较大的左心室尺寸，以及与盐水处理的对控剂相关的纤维化量更大。在存活手术期间保持无菌手术环境至关重要，以尽量减少术后感染和发病率。成功的模型创建可以通过连续超声心动图评估体内的肥大、扩张和功能障碍，以及通过组织学和分子评估收获的心脏组织进行外活。我们已将这种方法应用于100多种近亲繁殖小鼠，以评估由于慢性β-肾上腺素刺激导致的心脏重塑特征。结果公布并随时可用。

Research

JoVE Journal

Medicine

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

压力控制通气诱导小鼠急性肺损伤

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged ...

科研

医学

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

缺血性心力衰竭在临床前模型的转基因

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

植入小鼠触发剪应力致动脉粥样硬化的颈袖

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

The CYP2D6 Animal Model: How to Induce Autoimmune Hepatitis in Mice

CYP2D6动物模型：如何诱导小鼠自身免疫性肝炎

Autoimmune hepatitis is a rare but life threatening autoimmune disease of the liver of unknown etiology1,2. In the past many attempts have been made to ...

Implantation of the Syncardia Total Artificial Heart

SynCardia全人工心脏植入术

With advances in technology, the use of mechanical circulatory support devices for end stage heart failure has rapidly increased. The vast majority of ...

Implantation of Total Artificial Heart in Congenital Heart Disease

的全人工心脏的先天性心脏病植入

In patients with end-stage heart failure (HF), a total artificial heart (TAH) may be implanted as a bridge to cardiac transplant. However, in congenital ...

Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

左冠状动脉前降支的小鼠结扎：对心肌梗死后重塑和心衰模型

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

A Magnetic Microbead Occlusion Model to Induce Ocular Hypertension-Dependent Glaucoma in Mice

A磁条微珠缺血模型，以诱导小鼠高眼压，青光眼相关

The use of rodent models of glaucoma has been essential to understand the molecular mechanisms that underlie the pathophysiology of this multifactorial ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

球囊损伤诱发小鼠腹主动脉 Myointimal 增生的研究

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

Tachycardia-Induced Cardiomyopathy As a Chronic Heart Failure Model in Swine

心动过速诱发的心肌病作为猪慢性心力衰竭模型的研究

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment ...