Leducq基金会通过Caerus网络（RJH），支持这项工作是由美国国立卫生研究院R01 HL093183，HL088434，HL071763，HL080498，HL083156和P20HL100396（RJH）。 DL是由德国研究基金会的支持。我们要感谢她在这个项目的技术援助，劳伦Leonardson。

1。心肌梗死的创作<ol><li>女约克夏猪，动物生物技术产业（多伊尔斯敦，PA）（20KG）使用Telazol（tiletamine / zolazepam）（8.0毫克/公斤）和丁丙诺啡（0.6毫克）的前药。 </li><li>动物插管和100％的氧气，然后通风。整个过程中，全身麻醉维持异丙酚（5-8mg/kg/hr）。 </li><li>股的网站是准备用70％异丙醇，碘伏酒精。一个经皮穿刺提供访问动脉鞘的位置。鞘插入后，3000-5000 IU的肝素是管理四，维持活化凝血时间250-300秒。 </li><li>一个胺碘酮静脉推注（75毫克）的剂量肌肉注射。开始静脉注射胺碘酮（75毫克），阿托品（2毫克），醋酸钾（20毫克当量）1000毫升生理盐水袋和维护的整个过程中使用150毫升/小时的速度。 </li><li> 7神父曲棍球棒导管（Cordis公司;迈阿密，佛罗里达州）是先进的左冠状动脉和冠状动脉造影执行。一个0.014英寸的导丝（雅培，公园，IL）是先进到左冠状动脉前降支（LAD）。 8毫米长，4.0毫米旅行者过线的气球（雅培，公园，IL）是先进的，定位于近端的小伙子，和冠状动脉气球膨胀到3个大气压。法援署闭塞血管造影证实。 </li><li>随着连续的心电图和血流动力学监测，闭塞是保持了90分钟。恶性室性心律失常，偶尔会出现在第40分钟，在这种情况下，，直接从200 J 300焦耳的电流冲击即时交付管理。通常有两种心律失常的高峰时间帧，2至10分钟，球囊扩张后的25至35分钟。勤奋的监测和立即除颤心肌梗死成功创造的关键，因为有帧显示的时间内进入心室颤动的发生率在95％以上。如果执行的即时及适当的除颤，存活率非常高。如果在80毫米汞柱的血压下降，容积负荷是通过快速的静脉注射生理盐水输液。 </li><li>是瘪的气球和所有设备都将被删除。 </li><li>从股动脉鞘中删除，麻醉是功不可没的，恢复过程中的监控和动物。与胺碘酮，阿托品和醋酸钾输液静脉注射生理盐水50毫升/小时下降，是在一夜之间。 Nitroglycerline和Furoscemide肌肉注射给药。然后，动物被允许恢复和小心心电监护，持续至少3个小时。 </li><li>阿司匹林（200毫克）PO，将给予5天。 </li></ol>心肌梗死的创建过程，期间和之后的常见的并发症是冠状动脉解剖，穿刺部位血肿，心律失常，心力衰竭。一个完善的技术和全面监测，预防和管理这些并发症。 2。病毒注射<ol><li>动物准备，如心肌梗死创造steps1 - 3如上所述。应给予相同剂量的肝素，以防止血栓形成在病毒注射。 </li><li>一个5Fr曲棍球棒导管（Cordis公司;迈阿密，佛罗里达州）是先进的左冠状动脉。血管造影后，0.014英寸的导丝是先进的，到法援署到左回旋冠状动脉（LCX），以确定导管的位置。 </li><li>动脉血（-20毫升）是来自导管。为了防止缺血输液过程中，一半是用来稀释血液的解决方案。该病毒是用10毫升血混合，并用生理盐水稀释至20 ml。剩下的10毫升稀释至20 ml，并使用提供的导管管腔内残留的病毒。 </li><li>经耳缘静脉静脉硝酸甘油（20微克/分）开始。 </li><li>导管尖端放置在近端的左主道。通过导管的压力进行监控，以确保导管楔入。 </li><li>该病毒混合血（15毫升）是通过导管注入超过15分钟到左冠状动脉使用输液泵。 15分钟后，注射器交换刷新，和输液是持续10分钟。 </li><li>电线和导管固定在右冠状动脉，左冠状动脉的相同的方式。 </li><li>余下的5毫升血液病毒混合注射入右冠状动脉5分钟，冲洗10分钟。 </li></ol> 3。具有代表性的案例图1显示了在1个月后，心肌梗死创作的左心室。左室壁运动明显受损的射血分数（EF）31％。图2显示了在1个月的左心室的横截面。看到一个大的疤痕，从基地到前壁的心尖。在这个模型的心肌梗死后1个月的平均EF为36％和平均舒张末容积为83毫升，而在同一大小的动物无心肌梗死者的平均分别为68％和42毫升。图3显示了在左心室边境地带面积的表达β-半乳糖苷酶的腺相关病毒（2 × 10 12 DNA酶CMV LAC - Z的rAAV1 resitant颗粒）注射后1个月。增强的表达被发现的病毒注入面积。 <img src="/files/ftp_upload/2778/2778fig1.jpg" alt="图1" /> 图1。射血分数从左边心室心肌梗死创造后1个月。左心室扩张到92.5毫升和前壁的议案明显受损。 <img src="/files/ftp_upload/2778/2778fig2.jpg" alt="图2" /> 图2。氯化三苯四氮唑染色，在心肌梗死创造后1个月的左心室截面划定瘢痕区。看到一个大的疤痕，从基地到前壁的心尖。 <img src="/files/ftp_upload/2778/2778fig3.jpg" alt="图3" /> 图3β-半乳糖苷酶表达的腺相关病毒（2 × 10 12 DNA酶rAAV1 CMV LAC - Z的耐药颗粒）诱导。在边境地带面积梗死和冠状动脉内的病毒注射后的正常组织，1个月之间找到一个表达增强。右下图为阴性对照，只有生理盐水注入冠状动脉，使用相同的协议。

<ol>
	<li>Perez de Prado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Closed-chest+experimental+porcine+model+of+acute+myocardial+infarction-reperfusion.">Closed-chest experimental porcine model of acute myocardial infarction-reperfusion.</a> J Pharmacol Toxicol Methods. 60, 301-306 (2009).</li><li>Angeli, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Left+ventricular+remodeling+after+myocardial+infarction:+characterization+of+a+swine+model+on+beta-blocker+therapy.">Left ventricular remodeling after myocardial infarction: characterization of a swine model on beta-blocker therapy.</a> Comp Med. 59, 272-279 (2009).</li><li>Urdaneta, F., Lobato, E. B., Kirby, D. S. &. a. m. p. ;. a. m. p., Sidi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treating+myocardial+stunning+randomly,+with+either+propofol+or+isoflurane+following+transient+coronary+occlusion+and+reperfusion+in+pigs.">Treating myocardial stunning randomly, with either propofol or isoflurane following transient coronary occlusion and reperfusion in pigs.</a> Ann Card Anaesth. 12, 113-121 (2009).</li><li>Jaski, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium+upregulation+by+percutaneous+administration+of+gene+therapy+in+cardiac+disease+(CUPID+Trial),+a+first-in-human+phase+1/2+clinical+trial.">Calcium upregulation by percutaneous administration of gene therapy in cardiac disease (CUPID Trial), a first-in-human phase 1/2 clinical trial.</a> J Card Fail. 15, 171-181 (2009).</li></ol>

所有的作者有没有商业协会可能构成利益冲突，在手稿。

在各种原因引起的心力衰竭，缺血是由远在最常见的病因。为了克服这一严重的疾病，各种治疗方法正在制定中。需要仔细评估的疗法之前，这些有前途的疗法是可以应用到诊所。因此，建立的缺血性心力衰竭模型表示患者的临床情况，心力衰竭治疗的发展是至关重要的。我们的猪心肌梗死模型满足这些翻译的要求。 要通过创建一个心肌梗死诱发心力衰竭，心肌梗死面积的?...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>Telazol</td>
<td>&nbsp;</td>
<td>Fort Dodge Animal Health</td>
<td>0856-9050-49</td>
<td>&nbsp;</td>
<tr>
<td>Propofol</td>
<td>&nbsp;</td>
<td>APP Pharmaceuticals</td>
<td>63323-270-20</td>
<td>&nbsp;</td>
<tr>
<td>Heparin</td>
<td>&nbsp;</td>
<td>APP Pharmaceuticals</td>
<td>63323-540-11</td>
<td>&nbsp;</td>
<tr>
<td>Amiodarone</td>
<td>&nbsp;</td>
<td>APP Pharmaceuticals</td>
<td>63323-616-03</td>
<td>&nbsp;</td>
<tr>
<td>Atropin Sulfate</td>
<td>&nbsp;</td>
<td>Med-Pharmex</td>
<td>54925-063-10</td>
<td>&nbsp;</td>
<tr>
<td>Potassium acetate</td>
<td>&nbsp;</td>
<td>American Regent, Inc.</td>
<td>2053-25</td>
<td>&nbsp;</td>
<tr>
<td>Nitroglycerine</td>
<td>&nbsp;</td>
<td>American Regent, Inc.</td>
<td>4810-25 </td>
<td>&nbsp;</td>
<tr>
<td>Salix (Furosemide)</td>
<td>&nbsp;</td>
<td>Intervet Inc.</td>
<td>710461</td>
<td>&nbsp;</td>
<tr>
<td>5 Fr Hockey stick catheter</td>
<td>&nbsp;</td>
<td>Cordis</td>
<td>556-278-0L</td>
<td>&nbsp;</td>
<tr>
<td>7 Fr Hockey stick catheter</td>
<td>&nbsp;</td>
<td>Cordis</td>
<td>778-278-00</td>
<td>&nbsp;</td>
<tr>
<td>Coronary wire</td>
<td>&nbsp;</td>
<td>Abbott Laboratories</td>
<td>1044588</td>
<td>&nbsp;</td>
<tr>
<td>Coronary balloon</td>
<td>&nbsp;</td>
<td>Abbott Laboratories</td>
<td>1009447-08</td>
<td>&nbsp;</td>
<tr>
<td>8Fr sheath</td>
<td>&nbsp;</td>
<td>TERUMO</td>
<td>RSS801</td>
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gene transfer for ischemic heart failure in a preclinical model

各种新兴的技术正在发展为心衰患者。建立良好的临床前评估是必要的，以确定其疗效和安全性。 使用病毒载体的基因治疗是治疗心脏疾病的最有前途的的的方法之一。提供各种不同的基因的病毒载体基因通过改变具有不可估量的治疗潜力。 在这段视频中，基因转移创造心力衰竭的动物模型的全过程，提出了利用猪模型。第一，阻断近端的左冠状动脉前降支建立心肌梗死。慢性心功能不全心肌重塑的结果。我们的模型的独特之处是一个相当大的疤痕，真正体现严重心力衰竭的病人谁需要积极治疗取得积极成果。心肌梗死后瘢痕组织的创建和发展，同时硝化甘油输液内注射病毒表现。我们的注射方法提供了简单而有效的基因转移，基因表达增强。心肌梗死冠脉内传播病毒的猪模型的结合已被证明是一个一致的和可重复性的方法，这不仅有助于个人基因测试效果，但也比较的许多基因治疗候选人的疗效。

Watch this Scientific Journal Video about Gene Transfer for Ischemic Heart Failure in a Preclinical Model at JoVE.com

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

一个基因转染对缺血性心力衰竭的治疗方法是用一个猪心肌梗死模型。我们的简单和可重复性的方法，使我们能够很容易用一个非常简单和重复性的方式评估各种基因转移的疗效。

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 ...

gene-transfer-for-ischemic-heart-failure-in-a-preclinical-model

Research

JoVE Journal

Medicine

12.8K 次观看.  Mount Sinai School of Medicine. 一个基因转染对缺血性心力衰竭的治疗方法是用一个猪心肌梗死模型。我们的简单和可重复性的方法，使我们能够很容易用一个非常简单和重复性的方式评估各种基因转移的疗效。

视频: Gene Transfer for Ischemic Heart Failure in a Preclinical Model

A Simplified Technique for Producing an Ischemic Wound Model

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work well in non-ischemic wounds may not work in human diabetic wounds because vasculopathy is one major factor that hinders healing of these wounds. We published an article in 2007 describing a rabbit ear ischemic wound model created by a minimally invasive surgical technique. Since then, we have further simplified the procedure for easier operation. On one ear, three small skin incisions were made on the vascular pedicles, 1-2 cm from the ear base. The central artery was ligated and cut along with the nerve. The whole cranial bundle was cut and ligated, leaving only the caudal branch intact. A circumferential subcutaneous tunnel was made through the incisions, to cut subcutaneous tissues, muscles, nerves, and small vessels. The other ear was used as a non-ischemic control. Four wounds were made on the ventral side of each ear. This technique produces 4 ischemic wounds and 4 non-ischemic wounds in one animal for paired comparisons. After surgery, the ischemic ear was cool and cyanotic, and showed reduced movement and a lack of pulse in the ear artery. Skin temperature of the ischemic ear was 1-10 &deg;C lower than that on the normal ear and this difference was maintained for more than one month. Ear tissue high-energy phosphate contents were lower in the ischemic ear than the control ear. Wound healing times were longer in the ischemic ear than in the non-ischemic ear when the same treatment was used. The technique has now been used on more than 80 rabbits in which 23 were diabetic (diabetes time ranging from 2 weeks to 2 years). No single rabbit has developed any surgical complications such as bleeding, infection, or rupture in the skin incisions. The model has many advantages, such as little skin disruption, longer ischemic time, and higher success rate, when compared to many other models. It can be safely used in animals with reduced resistance, and can also be modified to meet different testing requirements.

We have developed a minimally invasive technique to create a rabbit ischemic ear wound model by dividing the central artery and nerve and the cranial neurovascular bundle. A subcutaneous tunnel then cuts all subcutaneous tissues. This procedure causes minimal skin disruption and can be safely used in diabetic animals.

一个生产缺血性损伤模型的简化技术

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work ...

科研

医学

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

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 is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

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 ...

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves focal endothelial-selective transgenesis, thereby allowing an investigator to determine the biological roles of endothelial-expressed transgenes and to quantify the in vivo transcriptional activity of DNA sequences in large artery endothelial cells. The method uses surgical isolation of rabbit common carotid arteries and an arteriotomy to deliver a transgene-expressing viral vector into the arterial lumen. A short incubation period of the vector in the lumen, with subsequent aspiration of the lumen contents, is sufficient to achieve efficient and durable expression of the transgene in the endothelium, with no detectable transduction or expression outside of the isolated arterial segment. The method allows assessment of the biological activities of transgene products both in normal arteries and in models of human vascular disease, while avoiding systemic effects that could be caused either by targeting gene delivery to other sites (e.g. the liver) or by the alternative approach of delivering genetic constructs to the endothelium by germ line transgenesis. Application of the method is limited by the need for a skilled surgeon and anesthetist, a well-equipped operating room, the costs of purchasing and housing rabbits, and the need for expertise in gene-transfer vector construction and use. Results obtained with this method include: transgene-related alterations in arterial structure, cellularity, extracellular matrix, or vasomotor function; increases or reductions in arterial inflammation; alterations in vascular cell apoptosis; and progression, retardation, or regression of diseases such as intimal hyperplasia or atherosclerosis. The method also allows measurement of the ability of native and synthetic DNA regulatory sequences to alter transgene expression in endothelial cells, providing results that include: levels of transgene mRNA, levels of transgene protein, and levels of transgene enzymatic activity.

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

This method is to introduce a transgene into the endothelium of rabbit carotid arteries. Introduction of the transgene allows the assessment of the biological role of the transgene product either in normal arteries or disease models. The method is also useful for measuring activity of DNA regulatory sequences.

体内兔颈总动脉内皮的基因转移

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves ...

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

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment methods. Here, we present such a model by tachycardia-induced cardiomyopathy, which can be produced by rapid cardiac pacing in swine.
A single pacing lead is introduced transvenously into fully anaesthetized healthy swine, to the apex of the right ventricle, and fixated. Its other end is then tunneled dorsally to the paravertebral region. There, it is connected to an in-house modified heart pacemaker unit that is then implanted in a subcutaneous pocket. 
After 4 - 8 weeks of rapid ventricular pacing at rates of 200 - 240 beats/min, physical examination revealed signs of severe heart failure - tachypnea, spontaneous sinus tachycardia, and fatigue. Echocardiography and X-ray showed dilation of all heart chambers, effusions, and severe systolic dysfunction. These findings correspond well to decompensated dilated cardiomyopathy and are also preserved after the cessation of pacing.
This model of tachycardia-induced cardiomyopathy can be used for studying the pathophysiology of progressive chronic heart failure, especially hemodynamic changes caused by new treatment modalities like mechanical circulatory supports. This methodology is easy to perform and the results are robust and reproducible.

Here, we present a protocol to produce tachycardia-induced cardiomyopathy in swine. This model represents a potent way to study the hemodynamics of progressive chronic heart failure and the effects of applied treatment.

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 ...

Isolation of Atrial Cardiomyocytes from a Rat Model of Metabolic Syndrome-related Heart Failure with Preserved Ejection Fraction

In this article, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes (ACMs) from a rat model of metabolic syndrome (MetS)-related heart failure with preserved ejection fraction (HFpEF). The prevalence of MetS-related HFpEF is rising, and atrial cardiomyopathies associated with atrial remodeling and atrial fibrillation are clinically highly relevant as atrial remodeling is an independent predictor of mortality. Studies with isolated single-cell cardiomyocytes are frequently used to corroborate and complement in vivo findings. Circulatory vessel rarefication and interstitial tissue fibrosis pose a potentially limiting factor for the successful single-cell isolation of ACMs from animal models of this disease.
We have addressed this issue by employing a device capable of manually regulating the intraluminal pressure of cardiac cavities during the isolation procedure, substantially increasing the yield of morphologically and functionally intact ACMs. The acquired cells can be used in a variety of different experiments, such as cell culture and functional Calcium imaging (i.e., excitation-contraction-coupling).
We provide the researcher with a step-by-step protocol, a list of optimized solutions, thorough instructions to prepare the necessary equipment, and a comprehensive troubleshooting guide. While the initial implementation of the procedure might be rather difficult, a successful adaptation will allow the reader to perform state-of-the-art ACM isolations in a rat model of MetS-related HFpEF for a broad spectrum of experiments.

Here, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes from a rat model of metabolic syndrome-related heart failure with preserved ejection fraction. A manual regulation of intraluminal pressure of cardiac cavities is implemented to yield functionally intact myocytes suitable for excitation-contraction-coupling studies.

代谢综合征心力衰竭模型大鼠心房心肌细胞的分离与保留射血分数的关系

In this article, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes (ACMs) from a rat model of ...

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The morphological and functional phenotyping of the right ventricle is of particular importance in the context of hemodynamic compromise in patients with ARHF. Here, we describe a method to induce ARHF in a previously described large animal model of chronic PH, and to phenotype, dynamically, right ventricular function using the gold standard method (i.e., pressure-volume PV loops) and with a non-invasive clinically available method (i.e., echocardiography). Chronic PH is first induced in pigs by left pulmonary artery ligation and right lower lobe embolism with biological glue once a week for 5 weeks. After 16 weeks, ARHF is induced by successive volume loading using saline followed by iterative pulmonary embolism until the ratio of the systolic pulmonary pressure over systemic pressure reaches 0.9 or until the systolic systemic pressure decreases below 90 mmHg. Hemodynamics are restored with dobutamine infusion (from 2.5 &#181;g/kg/min to 7.5 &#181;g/kg/min). PV-loops and echocardiography are performed during each condition. Each condition requires around 40 minutes for induction, hemodynamic stabilization and data acquisition. Out of 9 animals, 2 died immediately after pulmonary embolism and 7 completed the protocol, which illustrates the learning curve of the model. The model induced a 3-fold increase in mean pulmonary artery pressure. The PV-loop analysis showed that ventriculo-arterial coupling was preserved after volume loading, decreased after acute pulmonary embolism and was restored with dobutamine. Echocardiographic acquisitions allowed to quantify right ventricular parameters of morphology and function with good quality. We identified right ventricular ischemic lesions in the model. The model can be used to compare different treatments or to validate non-invasive parameters of right ventricular morphology and function in the context of ARHF.

We present a protocol to induce and phenotype an acute right heart failure in a large animal model with chronic pulmonary hypertension. This model can be used to test therapeutic interventions, to develop right heart metrics&#160;or to improve the understanding of acute right heart failure pathophysiology.

慢性血栓栓塞性肺动脉高压大型动物模型中急性右心衰竭的诱导和表型分析

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

糖尿病家兔欣德肢体缺血的临床前模型

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...

Establishing a Swine Model of Post-myocardial Infarction Heart Failure for Stem Cell Treatment

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major causes of mortality and morbidity around the world. Cell-based therapies for cardiac repair and improvement of left ventricular function after MI have attracted considerable attention. Accordingly, the safety and efficacy of these cell transplantations should be tested in a preclinical large animal model of HF prior to clinical use. Pigs are widely used for cardiovascular disease research due to their similarity to humans in terms of heart size and coronary anatomy. Therefore, we sought to present an effective protocol for the establishment of a porcine chronic HF model using closed-chest coronary balloon occlusion of the left circumflex artery (LCX), followed by rapid ventricular pacing induced with pacemaker implantation. Eight weeks later, the stem cells were administered by intramyocardial injection in the peri-infarct area. Then the infarct size, cell survival, and left ventricular function (including echocardiography, hemodynamic parameters, and electrophysiology) were evaluated. This study helps establish a stable preclinical large animal HF model for stem cell treatment.

We sought to establish a swine model of heart failure induced by left circumflex artery blockage and rapid pacing to test the effect and safety of intramyocardial administration of stem cells for cell-based therapies.

建立干细胞治疗心肌梗塞心力衰竭的猪模型

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major ...

A Preclinical Model of Exertional Heat Stroke in Mice

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, whereas exertional heat stroke (EHS) occurs during physical activity. EHS differs from CHS in etiology, clinical presentation, and sequelae of multi-organ dysfunction. Until recently, only models of CHS have been well established. This protocol aims to provide guidelines for a refined preclinical mouse model of EHS that is free from major limiting factors such as the use of anesthesia, restraint, rectal probes, or electric shock. Male and female C57Bl/6 mice, instrumented with core temperature (Tc) telemetric probes were utilized in this model. For familiarization with the running mode, mice undergo 3 weeks of training using both voluntary and forced running wheels. Thereafter, mice run on a forced wheel inside a climatic chamber set at 37.5 &#176;C and 40%-50% relative humidity (RH) until displaying symptom limitation (e.g., loss of consciousness) at Tc of 42.1-42.5 &#176;C, although suitable results can be obtained at chamber temperatures between 34.5-39.5 &#176;C and humidity between 30%-90%. Depending on the desired severity, mice are removed from the chamber immediately for recovery in ambient temperature or remain in the heated chamber for a longer duration, inducing a more severe exposure and a higher incidence of mortality. Results are compared with sham-matched exercise controls (EXC) and/or na&#239;ve controls (NC). The model mirrors many of the pathophysiological outcomes observed in human EHS, including loss of consciousness, severe hyperthermia, multi-organ damage as well as inflammatory cytokine release, and acute phase responses of the immune system. This model is ideal for hypothesis-driven research to test preventative and therapeutic strategies that may delay the onset of EHS or reduce the multi-organ damage that characterizes this manifestation.

The protocol describes the development of a standardized, repeatable, preclinical model of exertional heat stroke (EHS) in mice free from adverse external stimuli such as electric shock. The model provides a platform for mechanistic, preventative, and therapeutic studies.

小鼠劳力性中暑的临床前模型

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, ...

Large Animal Model for Evaluating the Efficacy of the Gene Therapy in Ischemic Heart

Coronary artery disease is one of the significant causes of mortality and morbidity worldwide. Despite the progression of current therapeutics, a considerable proportion of coronary artery disease patients remain symptomatic. Gene therapy-mediated therapeutic angiogenesis offers a novel therapeutic method for improving myocardial perfusion and relieving symptoms. Gene therapy with different angiogenic factors has been studied in few clinical trials. Due to the novelty of the method, the progress of myocardial gene therapy is a continuous path from bench to bedside. Therefore, large animal models are needed for evaluating the safety and efficacy. The more the large animal model identifies the original disease and the endpoints used in clinics, the more predictable outcomes are from clinical trials. Here, we introduce a large animal model for evaluating the efficacy of the gene therapy in the ischemic porcine heart. We use clinically relevant imaging methods such as ultrasound imaging and 15H2O-PET. For targeting the gene transfers into the desired area, electroanatomical mapping is used. The aim of this method is: (1) to mimic chronic coronary artery disease, (2) to induce therapeutic angiogenesis at hypoxic areas of the heart, and (3) to evaluate the safety and efficacy of the gene therapy by using relevant endpoints.

Myocardial gene therapy for ischemic heart disease holds great promise for future therapeutics. Here, we introduce a large animal model for evaluating the efficacy of gene therapy in the ischemic heart.