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).

Procedures waarbij proefdieren zijn goedgekeurd door de Nederlandse Centrale Commissie Dierproeven en de Animal Care Comite op Universitair Medisch Centrum Groningen (NL). Zowel Wistar en Lewis ratten met gewichten tussen 180 en 300 g werden gebruikt. 1. Huisvesting en acclimatisatie <ol><li> Na aankomst bij de centrale dier faciliteit, huis ratten in groepen van 5 per kooi. Gedurende een 7-dagen acclimatisatie periode wennen de ratten humane behandeling, maar geen experimentele procedures...

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Deze methode beschrijft de chirurgische procedure van een aorto-caval shunt bij ratten voorbehandeld met MCT tot stromingsgeïnduceerde PAK en de technieken om het beginsel van hemodynamische en histopathologische eindpunten dat PAK en dit model te karakteriseren beoordelen creëren. Kritische stappen in het protocol en problemen oplossen Chirurgie en na de operatie. Tijdens de aorto- cavale shunt operatie, de meest kritische stap is de dissectie van de aorta en ven...

Het doel van deze methode is om een ​​reproduceerbaar model voor ernstige stromingsgeïnduceerde pulmonale arteriële hypertensie in ratten maken en dat in beginsel hemodynamische en histopathologische eindpunten meten. Pulmonale arteriële hypertensie (PAH) is een klinisch syndroom dat een geleidelijke verhoging van de pulmonale vasculaire weerstand leidt tot rechter ventrikel falen en dood omvat. Binnen het overkoepelende ziektespectrum van pulmonaire hypertensieve ziekten (PH), PAH is de ernstigste vorm en die zonder behandeling 1 blijft. De onderliggende arteriopathie in PAH wordt gekenmerkt door een typische vorm van vasculaire hermodellering dat het vatlumen afsluit. Muscularisatie van normale niet-gemusculariseerd schepen en hypertrofie van de mediale lagen vat worden beschouwd reeds ziekteverschijnselen in PAH ook gezien in andere vormen van PH 2 en 3 worden gedacht omkeerbaar zijn. Als PAH eendvances, de intima-laag begint te verbouwen, uiteindelijk de vorming van karakteristieke neointima laesies 2. Nieuwe intima-type vasculaire longremodellering is exclusief voor PAH en wordt momenteel beschouwd onomkeerbaar 4 te zijn. Als PAH is een zeldzame ziekte, de vooruitgang in de pathobiological begrip en de ontwikkeling van nieuwe therapieën hebben zwaar op diermodellen ingeroepen. De monocrotalin (MCT) model in ratten een single hit model is, en nog steeds vaak gebruikt. MCT is een toxine dat de schade toebrengt aan de pulmonale arteriolen en regionale ontsteking 5. 60 mg / kg MCT leidt tot een verhoging van de gemiddelde pulmonale arteriële druk (mPAP), pulmonale vasculaire weerstand (PVR) en rechterventrikelhypertrofie (RVH) na 3-4 weken 6. De histomorphology wordt gekenmerkt door geïsoleerde mediale hypertrofie zonder neointima laesies 5. de MCTrattenmodel vormt derhalve een matige vorm van PH en niet PAK, hoewel het gewoonlijk wordt voorgesteld als de laatste. Bij kinderen met PAH geassocieerd met congenitale links naar rechts shunt (PAH-CHD), verhoogde pulmonaire bloedstroom wordt beschouwd als de essentiële trigger voor de ontwikkeling van neointima laesies 7, 8, 9. Bij ratten kunnen verhoogde pulmonaire bloedstroom worden geïnduceerd door het creëren van een shunt tussen de abdominale aorta en vena cava, een techniek die het eerst beschreven in 1990 10. Alternatieven voor verhoogde pulmonale stroom te creëren zijn door eenzijdige pneumonectomie of subclavia naar longslagader anastomose 11. Conceptuele nadelen van deze modellen bestaan ​​potentiële compenserende groei van de overgebleven long en adaptieve pathway activatie geïnduceerd door de pneumonectomie of iatrogene schade van de pulmonaire vasculatuur te wijtenom longslagader anastomose, zowel verstorende effecten van de toegenomen pulmonale bloedstroom. Wanneer een aorto- cavale shunt gecreëerd en verhoogde pulmonale bloedstroom wordt geïnduceerd als tweede hit in MCT behandelde ratten, origineel neointima laesies ontstaan ​​en een ernstige vorm van PAH en bijbehorende rechterkamer falen (RVF) ontwikkelen 3 weken na de toegenomen stromen 12. De hemodynamische progressie van PAK&#39;s in dit model kan in vivo worden beoordeeld door echocardiografie en rechter hartkatheterisatie. Het vasculaire histomorphology, wanddikte van het drukvat, de mate van arteriolaire occlusie, en parameters voor de rechter ventrikel falen vormen de pijlers van het ex vivo karakterisering van PAH. Deze methode beschrijft gedetailleerde protocollen voor de aorto-cavale shunt (AC-shunt) chirurgie, rechts hartkatheterisatie, en de kwalitatieve en kwantitatieve beoordeling van vasculaire histomorphology.

<table border="0" cellpadding="0" cellspacing="0" width="835">
	<tbody>
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			<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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		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Scalple blade</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;">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.

Representatieve resultaten zijn weergegeven in figuur 4. De gepresenteerde resultaten tonen kenmerken van MCT + FLOW in Lewis ratten in de volgende groepen: Controle (n = 3), MF8 (n = 5), MF14 (n = 5), MF28 (n = 5) en MF-RVF ( n = 10). Statistische analyses werden uitgevoerd met de eenzijdige ANOVA met Bonferroni-correctie. 60 mg / kg MCT en verhoogde pulmonaire bloedstroom leiden tot een gemiddelde stijging v...

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.

Shunt Chirurgie, Right hartkatheterisatie, en Vasculaire Morphometry in een diermodel voor Flow-geïnduceerde pulmonale arteriële hypertensie

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

Femorale arteriële en veneuze catheterisatie voor bloedafname, Drug Administration en bewust Bloeddruk en hartslag metingen

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

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

Chirurgische ingrepen voor een Rat Model van deelspoelen orthotope levertransplantatie met hepatische arteriële reconstructie

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

Echocardiografische Beoordeling van de rechter hart in Muizen

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

Chronische trombo-embolische pulmonale hypertensie en beoordeling van rechter ventrikel functie in de 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

Hemodynamische karakterisatie van diermodellen van pulmonale arteriële hypertensie

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

Inductie en fenotypering van acuut rechterhartfalen in een groot diermodel van chronische trombo-embolische pulmonale hypertensie

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

Linker atriale stenose geïnduceerde pulmonale veneuze Arterialization en groep 2 pulmonale hypertensie in Rat

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

Invasieve hemodynamische beoordeling voor het rechter ventrikel systeem en hypoxie-geïnduceerde pulmonale arteriële hypertensie bij muizen

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

Een Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury met Periadventitial Therapeutic Application

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