Name: induction and assessment of ischemia-reperfusion injury in langendorff-perfused rat hearts
Uploaded: 2015-07-27T15:00:00
Description: Watch this Scientific Journal Video about Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts at JoVE.com

Bu yayın Güney Carolina Tıp Üniversitesi akademik ev, NIH / NCATS Hibe Numarası UL1 TR000062 ile, South Carolina Klinik ve Translational Research (SCTR) Enstitüsü tarafından desteklenmiştir. Ek destek DRM VA liyakat ödülü BX002327-01 tarafından sağlandı. DJH NIH / NCATS Hibe Numarası TL1 TR000061 tarafından ve NIH Hibe Numarası T32 GM008716 tarafından desteklenmiştir. DENİZ NIH Hibe Numarası T32 HL07260 tarafından desteklenmiştir.

Burada listelenen tüm işlemler Güney Carolina Tıp Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından onaylanmıştır. Burada tarif edilen deneyler, akut, hayatta olmayan deneylerdir. Bu nedenle, var göz merhemi hiçbir kullanımı ve steril bir çalışma paketi gerekli değildir. Ötanazi kalbin hasat sırasında eksanguinasyon yoluyla elde edilir. 1. Deneysel hazırlanması <ol><li> Sabit bir basınç veya sabit akış Langendorff perfüzyon Aparatı 4 ...

<ol>
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(65), (2012).</li><li>Sill, B., Hammer, P. E., Cowan, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+mapping+of+langendorff-perfused+rat+hearts.">Optical mapping of langendorff-perfused rat hearts.</a> The Journal of Visualized Experiments. (30), (2009).</li><li>Ferrera, R., Benhabbouche, S., Bopassa, J. C., Li, B., Ovize, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=One+hour+reperfusion+is+enough+to+assess+function+and+infarct+size+with+TTC+staining+in+langendorff+rat+model.">One hour reperfusion is enough to assess function and infarct size with TTC staining in langendorff rat model.</a> Cardiovascular Drugs and Therapy. 23 (4), 327-331 (2009).</li><li>Sutherland, F. J., Shattock, M. J., Baker, K. E., Hearse, D. 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İzole perfüze sıçan kalp başarıyla iskemi reperfüzyon hasarı 9 kardiyak önkoşullama üzerindeki farmakolojik müdahalenin etkisini araştırmak için kullanılabilir. Ancak, tekrarlanabilir sonuçlar sağlamak için standardize edilmelidir prosedüre bazı temel adımlar vardır. Sistem içinde 37.4 ° C arasında bir sıcaklık muhafaza hatta hafif ve hipotermi hipertermi kalp önkoşullanma 10,11 neden olabilir, kritik öneme sahiptir. ketamin ile uzun süreli maruz kalp önkoşullama <...

Hem iskemi ve reperfüzyon kardiyak tepkiyi altında yatan olaylar izahı aort kros-sıkma 2. gerektiren miyokard infarktüsü 1 ve kardiyak cerrahi işlemlerin tedavi geliştirilmesi esastır. Iskemi reperfüzyon hasarının in vivo modelleri, çok kullanışlı bir son nokta analizi sağlarken, gerçek zamanlı olarak, akut iskemi reperfüzyon hasarının fonksiyonel etkilerinin araştırılması için etkili değildir. In vivo iskemi reperfüzyon modelleri genellikle reperfüzyon sırasında infarkt önemli değişkenlik ve kalbe ilacın doğrudan teslim üretmek içinde Ayrıca, zordur. iskemi reperfüzyon hasarının incelenmesi için bir Langendorff izole kalp sisteminin kullanımı, farmakolojik tedaviler, enfarkte doku tek biçimli alan ve doğrudan miyokarda ilacın anlık teslimat gerçek zamanlı fonksiyonel değerlendirmesi için izin verir. İlk b&#39;deki1895, 3 y Oscar Langendorff, Langendorff izole kalp son 40 yıl 4,5 için iskemi reperfüzyon araştırmalarında kullanılan edilerek, iskemi reperfüzyon hasarı incelemek için sağlam bir modeldir. Burada, bazı değişiklikler fonksiyonel analiz için izole kalbi optimize etmek için yapılır. Aortun yerinde kanülasyon kalp kalpte iskemi reperfüzyon çalışmaların 6 sonuçlarını değiştirecek iskemik önkoşullanma, tecrübe etmediğini garanti iken. Bunu kolaylaştırmak için, bir traketomi havalandırma sağlayan ve ameliyat sırasında sıçan fizyolojik istikrarın sağlanması gerçekleştirilir. Bundan sonra, kalp, Krebs Henseleit tamponu aort içine doğrudan retrograd perfüzyon yoluyla teslim edildiği bir cam su ceketli sarmal kolonuna bağlanmıştır. Bir tuzlu su dolu balon sol ventrikül içerisine eklenen ve ventrikül An içinden baskıların gerçek zamanlı ölçümüne izin veren bir basınç transdüktörüne, bağlı olduğuBirden fazla fonksiyonel parametreler d hesaplanması. Deneyin sonunda, kalp kasılma ve DNA, RNA ve protein seviyeleri alt analiz sağlamak için sıvı azot içinde dondurulmuş flaş tutulması için bir soğuk tuzlu su ile yıkanır. Böylece değiştirilmiş, Langendorff perfüze kalp iskemi reperfüzyon hasarı sırasında akut herhangi bir zamanda farmakolojik müdahalelerin fizyolojik etkisinin direkt izlenmesi için etkili bir sistem olarak hizmet vermektedir.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Sodium Chloride</td>
			<td>Sigma Aldrich</td>
			<td>S3014</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium Chloride</td>
			<td>Sigma Aldrich</td>
			<td>P9541</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium Sulfate</td>
			<td>Sigma Aldrich</td>
			<td align="right">203726</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium Phosphate Dibasic</td>
			<td>Sigma Aldrich</td>
			<td>RES20765-A7</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium Chloride Dihydrate</td>
			<td>Sigma Aldrich</td>
			<td>C8106</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Bicarbonate</td>
			<td>Sigma Aldrich</td>
			<td>S5761</td>
			<td></td>
		</tr>
		<tr>
			<td>D-Glucose</td>
			<td>Sigma Aldrich</td>
			<td>G8270</td>
			<td></td>
		</tr>
		<tr>
			<td>Octanoic Acid</td>
			<td>Sigma Aldrich</td>
			<td>C2875</td>
			<td></td>
		</tr>
		<tr>
			<td>2,3,5-triphenyltetrazolium chloride</td>
			<td>Sigma Aldrich</td>
			<td>T8877</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical Pressure Transducer</td>
			<td>MEMSCAP</td>
			<td>SP844</td>
			<td></td>
		</tr>
		<tr>
			<td>Masterflex Peristaltic Pump</td>
			<td>Cole Parmer</td>
			<td>EW-07521-40</td>
			<td></td>
		</tr>
		<tr>
			<td>Masterflex Easy Load Pump Head</td>
			<td>Cole Parmer</td>
			<td>EW-07518-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Heated circulating water bath</td>
			<td>Lauda</td>
			<td>M3</td>
			<td></td>
		</tr>
		<tr>
			<td>Tubing Flow Module</td>
			<td>Transonic</td>
			<td>TS410</td>
			<td></td>
		</tr>
		<tr>
			<td>Modular Research Console</td>
			<td>Transonic</td>
			<td>T402</td>
			<td></td>
		</tr>
		<tr>
			<td>Inline flow sensor</td>
			<td>Transonic</td>
			<td>ME2PXN</td>
			<td></td>
		</tr>
		<tr>
			<td>PowerLab Data Acquisition Device</td>
			<td>AD Instruments</td>
			<td>PL3508</td>
			<td></td>
		</tr>
		<tr>
			<td>LabChart data acquisition software</td>
			<td>AD Instruments</td>
			<td>MLU60/8</td>
			<td></td>
		</tr>
	</tbody>
</table>

induction and assessment of ischemia-reperfusion injury in langendorff-perfused rat hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for myocardial infarction and cardiac complications of thoracic surgery. The ability of certain drugs to precondition the myocardium against ischemia reperfusion injury has led to multiple clinical trials, with little success. The isolated heart model allows acute observation of the functional effects of ischemia reperfusion injury in real time, including the effects of various pharmacological interventions administered at any time-point before or within the ischemia-reperfusion injury window. Since brief periods of ischemia can precondition the heart against ischemic injury, in situ aortic cannulation is performed to allow for functional assessment of non-preconditioned myocardium. A saline filled balloon is placed into the left ventricle to allow for real-time measurement of pressure generation. Ischemic injury is simulated by the cessation of perfusion buffer flow, followed by reperfusion. The duration of both ischemia and reperfusion can be modulated to examine biochemical events at any given time-point. Although the Langendorff isolated heart model does not allow for the consideration of systemic events affecting ischemia and reperfusion, it is an excellent model for the examination of acute functional and biochemical events within the window of ischemia reperfusion injury as well as the effect of pharmacological intervention on cardiac pre- and postconditioning. The goal of this protocol is to demonstrate how to perform in situ aortic cannulation and heart excision followed by ischemia/reperfusion injury in the Langendorff model.

sol ventrikül balon aparatı müteahhitlik sol ventrikül (Şekil 1) tarafından geliştirilen basınç gerçek zamanlı izleme sağlar. Daha önce 7 tarif edildiği gibi, bu basınç izleme ventriküler fonksiyon parametrelerinin çoğu hesaplamak için kullanılabilir. Bu hesaplamalar, başlangıç ​​aşaması olarak reperfüzyon fazı yapılan her bir grup içinde birden fazla izleri üzerinden ortalaması alınmış, ve daha önce 9 yapmış gibi farmakolojik müdahale, ka...

Watch this Scientific Journal Video about Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts at JoVE.com

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts (Video) | JoVE

The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

Langendorff-perfüze sıçan Hearts indüksiyon ve İskemi-reperfüzyon Hasarı Değerlendirilmesi

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

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Medicine

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

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Bağışıklık Yanıtları intravital multiphoton Görüntüleme Fare Kulak Skin uyaran İskemi-reperfüzyon yaralanması

Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent ...

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

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

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