Name: shunt surgery, right heart catheterization, and vascular morphometry in a rat model for flow-induced pulmonary arterial hypertension
Uploaded: 2017-02-11T13:00:00
Description: Watch this Scientific Journal Video about Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension at JoVE.com

This study was supported by the Netherlands Cardiovascular Research Initiative, the Dutch Heart Foundation, the Dutch Federation of University Medical Centers, the Netherlands Organization for Health Research and Development, and the Royal Netherlands Academy of Sciences (CVON nr. 2012-08, PHAEDRA, The Sebald fund, Stichting Hartekind).

Procedimentos envolvendo indivíduos animais foram aprovados pelo Comité Central Holandês para Experimentação Animal e do Comité de Cuidados com Animais da Universidade Medical Center Groningen (NL). foram usados ​​tanto em ratos Wistar e Lewis com pesos entre 180 e 300 g. 1. Habitação e aclimatação <ol><li> Depois da chegada ao biotério central, ratos de casas em grupos de 5 por gaiola. Durante um período de aclimatação de 7 dias, acostumar os ratos a manipulação humana...

<ol>
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Este método descreve o procedimento cirúrgico de shunt aorto-cava em ratos pré-tratados com MCT para criar HAP induzida pelo fluxo e as técnicas para avaliar o princípio hemodinâmica e pontos finais histopatológicos que caracterizam HAP e este modelo. Passos críticos dentro do Protocolo e solução de problemas Cirurgia e pós-cirurgia. Durante a cirurgia de derivação aorto-cava, a etapa mais crítica é a dissecção da aorta e veia cava. As membranas que...

O objetivo desse método é criar um modelo reproduzível para a hipertensão arterial pulmonar grave, induzida por fluxo em ratos e medir o seu princípio hemodinâmica e pontos finais histopatológicos. A hipertensão arterial pulmonar (HAP) é uma síndrome clínica que engloba um aumento progressivo na resistência vascular pulmonar, levando à falência ventricular direita e morte. Dentro do espectro da doença subordinador de doenças de hipertensão pulmonar (PH), HAP é a forma mais grave e uma que permanece sem cura 1. A arteriopatia subjacente na HAP é caracterizada por uma forma típica de remodelação vascular que provoca a oclusão do lúmen do vaso. Muscularização de navios não muscularização normais e hipertrofia da camada navio medial são considerados fenômenos precoce da doença em HAP, também são vistos em outras formas de PH 2, e são pensados para ser reversível 3. Conforme a HAP umdvances, a camada íntima começa a remodelar, eventualmente formando lesões neointimais característicos 2. -Type neointimal remodelamento vascular pulmonar é exclusivo para PAH e está actualmente a ser considerada irreversível 4. Conforme a HAP é uma doença rara, os avanços na sua compreensão pathobiological e desenvolvimento de novas terapias têm se baseou fortemente em modelos animais. O modelo monocrotalin (MCT) em ratos é um simples modelo de hit single que tem sido, e ainda é, frequentemente utilizado. MCT é uma toxina que causa lesão aos arteríolas pulmonares e inflamação regionais 5. 60 mg / kg de MCT leva a um aumento da pressão arterial pulmonar média (PAP), a resistência vascular pulmonar (RVP), e a hipertrofia ventricular direita (HVD) depois de 3 - 4 semanas 6. O histomorfologia é caracterizada por hipertrofia medial isolada, sem lesões neointimais 5. o MCTmodelo de rato representa, assim, uma forma moderada de PH, e não HAP, embora seja normalmente apresentado como o último. Em crianças com HAP associada a um shunt congênita da esquerda para a direita (HAP-CHD), aumento do fluxo sanguíneo pulmonar é considerado como o gatilho essencial para o desenvolvimento de lesões neointimais 7, 8, 9. Em ratos, o aumento do fluxo sanguíneo pulmonar pode ser induzida através da criação de uma derivação entre a aorta abdominal e a veia cava, uma técnica descrita pela primeira vez em 1990 10. Alternativas para criar aumento do fluxo pulmonar são por pneumonectomia unilateral ou subclávia à anastomose da artéria pulmonar 11. desvantagens conceituais desses modelos consistem em potencial de crescimento compensatório do pulmão remanescente e ativação da via de adaptação induzida pela pneumonectomia, ou de lesão iatrogênica da vasculatura pulmonar devidoa anastomose da artéria pulmonar, tanto confundindo os efeitos do aumento do fluxo sanguíneo pulmonar. Quando um shunt aorto-cava é criado e aumento do fluxo sanguíneo pulmonar é induzida como um segundo hit em ratos tratados com MCT, lesões neointimais característicos ocorrem, e uma forma grave de HAP e falência do ventrículo direito associada (RVF) Desenvolver 3 semanas após o aumento fluir 12. A progressão hemodinâmica da HAP neste modelo pode ser avaliada in vivo pelo ecocardiograma e cateterismo cardíaco direito. O histomorfologia vascular, espessura da parede do vaso, o grau de oclusão arteriolar, e os parâmetros para a falência do ventrículo direito formam os pilares da caracterização ex vivo da HAP. Este método descreve protocolos detalhados para a cirurgia de derivação aorto-cava (AC-shunt), cateterismo cardíaco direito e avaliação qualitativa e quantitativa de histomorfologia vascular.

<table border="0" cellpadding="0" cellspacing="0" width="835">
	<tbody>
		<tr height="25">
			<td height="25" style="height:25px;width:307px;">
			Shunt Surgery
			</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td style="width:196px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sterile surgical gloves</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Duratears Eye ointment</td>
			<td style="width:151px;">Alcon</td>
			<td style="width:183px;">10380</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Chloride-Hexidine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Cotton swabs</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Histoacryllic tissue glue</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td>1050052</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Silkam 5-0 sutures black non-resorbable</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td>F1134027</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Safil 4-0 sutures violet resorbable</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">18 G needle&#160;</td>
			<td style="width:151px;">Luer</td>
			<td>NN1838R BD</td>
			<td>tip bent in 45 degrees orifice to the outside</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Gauzes 10x10 cm</td>
			<td>Paul Hartmann</td>
			<td>407825</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Temgesic Buprenorphine</td>
			<td style="width:151px;">RB Pharmaceuticals</td>
			<td style="width:183px;">5429</td>
			<td>subcutaneous injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sodium Chloride 0.9 %</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Ventilation mask Rat</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:307px;">Scalple blade</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Biemer clamp 18 mm, 5 mm opening&#160;</td>
			<td><a href="https://www.google.nl/url?sa=i&amp;rct=j&amp;q=&amp;esrc=s&amp;source=images&amp;cd=&amp;cad=rja&amp;uact=8&amp;ved=0ahUKEwitqICpkITNAhWiL8AKHe6iCC8QjhwIBQ&amp;url=http%3A%2F%2Fwww.agnthos.se%2Fquote%2Fquote%2Fcaturl%2Fid%2F242%2FBiemer&amp;psig=AFQjCNEnfRXuMQ6Doi4_-2KMYu3qeRovMA&amp;ust=1464777707579746">AgnTho</a></td>
			<td>64-562</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Heat mat</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Kocher Clamp</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:307px;">Shaving machine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Microscope</td>
			<td style="width:151px;">Leica</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:307px;">
			Right Heart Catheterization
			</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Name</td>
			<td style="width:151px;">Company</td>
			<td style="width:183px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sterile surgical gloves</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Eye ointment</td>
			<td style="width:151px;">Duratears</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Chloride-Hexidine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Cotton swabs</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Gauzes 10x10 cm</td>
			<td>Paul Hartmann</td>
			<td>407825</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Silkam 5-0 sutures black non-resorbable</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td>F1134027</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Needle 20 G</td>
			<td style="width:151px;">Luer</td>
			<td style="width:183px;"></td>
			<td>Tip slightly bent to the inside</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Cannula 20 G</td>
			<td style="width:151px;">Luer</td>
			<td style="width:183px;"></td>
			<td>to introduce catheter, tip pre-formed in 20 degrees</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Silastic Catheter 15 cm long</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td>0.5 mm ball 2 mm from tip</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Pressure transducer</td>
			<td style="width:151px;">Ailtech</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Bedside monitor Cardiocap/5</td>
			<td style="width:151px;">Datex-Ohmeda</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Shaving machine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">10mL Syringe</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sodium Chloride 0.9 %</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td>for flushing</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:307px;">
			Vascular Morphology
			</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Name</td>
			<td style="width:151px;">Company</td>
			<td style="width:183px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">50ml Syringe</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">4 % Formaldehyde</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">18 G cannula with tube</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Verhoef staining kit</td>
			<td style="width:151px;">Sigma-Aldrich</td>
			<td><a href="http://www.sigmaaldrich.com/catalog/product/sigma/ht254?lang=en&amp;region=NL">HT254</a></td>
			<td>http://www.sigmaaldrich.com/catalog/product/sigma/ht254?lang=en&#38;region=US</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Digital slide scanner</td>
			<td style="width:151px;">Hamamatsu</td>
			<td style="width:183px;">C9600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Image-J</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Elastic (Connective Tissue Stain)&#160;</td>
			<td style="width:151px;">Abcam</td>
			<td style="width:183px;">ab150667</td>
			<td>http://www.abcam.com/elastic-connective-tissue-stain-ab150667.html</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;"></td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td>http://www.abcam.com/ps/products/150/ab150667/documents/ab150667-Elastic%20Stain%20Kit%20(website).pdf</td>
		</tr>
	</tbody>
</table>

shunt surgery, right heart catheterization, and vascular morphometry in a rat model for flow-induced pulmonary arterial hypertension

In this protocol, PAH is induced by combining a 60 mg/kg monocrotalin (MCT) injection with increased pulmonary blood flow through an aorto-caval shunt (MCT+Flow). The shunt is created by inserting an 18-G needle from the abdominal aorta into the adjacent caval vein. Increased pulmonary flow has been demonstrated as an essential trigger for a severe form of PAH with distinct phases of disease progression, characterized by early medial hypertrophy followed by neointimal lesions and the progressive occlusion of the small pulmonary vessels. To measure the right heart and pulmonary hemodynamics in this model, right heart catheterization is performed by inserting a rigid cannula containing a flexible ball-tip catheter via the right jugular vein into the right ventricle. The catheter is then advanced into the main and the more distal pulmonary arteries. The histopathology of the pulmonary vasculature is assessed qualitatively, by scoring the pre- and intra-acinar vessels on the degree of muscularization and the presence of a neointima, and quantitatively, by measuring the wall thickness, the wall-lumen ratios, and the occlusion score.

Os resultados representativos são apresentados na Figura 4. Os resultados apresentados mostram características do MCT + FLUXO em ratos Lewis nos seguintes grupos: Controle (n = 3), MF8 (n = 5), MF14 (n = 5), MF28 (n = 5) e MF-RVF ( n = 10). As análises estatísticas foram realizadas utilizando o one-way ANOVA com correção de Bonferroni. 60 mg / kg MCT e aumentou a vantagem fluxo sanguíneo pulmonar a um a...

Watch this Scientific Journal Video about Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension at JoVE.com

Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension

This protocol describes a surgical procedure to create a model for flow-induced pulmonary arterial hypertension (PAH) in rats and the procedures to analyze the principle hemodynamic and histological end-points in this model.

A cirurgia de derivação, coração direito Cateterismo e Vascular morfometria em um modelo de rato para induzida Fluxo de Hipertensão Arterial Pulmonar

In this protocol, PAH is induced by combining a 60 mg/kg monocrotalin (MCT) injection with increased pulmonary blood flow through an aorto-caval shunt ...

Research

JoVE Journal

Medicine

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

Cateterismo arterial e venoso femoral para coleta de sangue, Drug Administration e Pressão Arterial Consciente e Medidas de freqüência cardíaca

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

Medicina

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Procedimentos cirúrgicos para um modelo de rato de Transplante de Fígado parcial ortotópico com reconstrução arterial hepática

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Echocardiographic Assessment of the Right Heart in Mice

Avaliação ecocardiográfica do coração direito em Ratos

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

Chronic Thromboembolic Pulmonary Hypertension and Assessment of Right Ventricular Function in the Piglet

Tromboembólica crônica Hipertensão Pulmonar e Avaliação da Função Ventricular Direita no Piglet

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. ...

Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension

Hemodinâmica Caracterização de roedores modelos de Hipertensão Arterial Pulmonar

Pulmonary arterial hypertension (PAH) is a rare disease of the pulmonary vasculature characterized by endothelial cell apoptosis, smooth muscle ...

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

Indução e Fenotipagem de Insuficiência Cardíaca Aguda Direita em um Grande Modelo Animal de Hipertensão Pulmonar Tromboembolic Crônica

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

Left Atrial Stenosis Induced Pulmonary Venous Arterialization and Group 2 Pulmonary Hypertension in Rat

Estenose Atrial Esquerdo induzida Arterialization venoso pulmonar e hipertensão pulmonar do grupo 2 no rato

The mechanism of mitral stenosis-induced pulmonary venous arterialization and group 2 pulmonary hypertension (PH) is unclear. There is no rodent model of ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Avaliação Hemodinâmica Invasiva para o Sistema Ventricular Direito e hipertensão arterial pulmonar induzida por hipóxia em camundongos

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Uma lesão de balão segmental controlada por pressão da artéria de rato com aplicação terapêutica periadventicional

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...