Name: hemodynamic characterization of rodent models of pulmonary arterial hypertension
Uploaded: 2016-04-11T14:00:00
Description: Watch this Scientific Journal Video about Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension at JoVE.com

SR is supported by NIH K08HL114643, Gilead Research Scholars in Pulmonary Arterial Hypertension and a Burroughs Wellcome Fund Career Award for Medical Scientists.

Tutte le procedure descritte seguono le linee guida per la cura degli animali della Duke University School of Medicine. 1. Prima di iniziare la procedura Nota: prima di qualsiasi procedura di animali, in modo che l&#39;autorizzazione istituzionale adeguato è stato ottenuto. Come per tutte le procedure, utilizzare appropriate antidolorifici per assicurare che non ci sia la sofferenza degli animali. <ol><li> cateteri a filo con soluzione salina sterile eparinizz...

<ol>
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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=Bisoprolol+delays+progression+towards+right+heart+failure+in+experimental+pulmonary+hypertension.">Bisoprolol delays progression towards right heart failure in experimental pulmonary hypertension.</a> Circ Heart Fail. 5, 97-105 (2012).</li><li>de Man, F. 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=Dysregulated+renin-angiotensin-aldosterone+system+contributes+to+pulmonary+arterial+hypertension.">Dysregulated renin-angiotensin-aldosterone system contributes to pulmonary arterial hypertension.</a> Am J Respir Crit Care Med. 186, 780-789 (2012).</li><li>Pritts, C. D., Pearl, R. 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The protocols outlined here describe a comprehensive characterization of hemodynamics and right ventricular function in rodent models of pulmonary hypertension. While right heart catheterization as described here is a terminal procedure, the mortality associated with echocardiography is minimal, which allows for screening and follow-up of disease progression. However, similar to patients with PH having markedly increased mortality with anesthesia17, in our experience, rats with severe PH do not tolerate anesth...

L&#39;ipertensione arteriosa polmonare (PAH) è una malattia del sistema vascolare polmonare associata a infiltrazione di cellule infiammatorie, la proliferazione della muscolatura liscia e l&#39;apoptosi delle cellule endoteliali. Questi cambiamenti si traducono in obliterazione delle arteriole polmonari, successivamente portando a ventricolo destro (RV) disfunzione e insufficienza cardiaca. Al fine di comprendere la fisiopatologia sottostante PAH e insufficienza RV in PAH, sono stati sviluppati una serie di diversi modelli, tra cui modelli genetici e farmacologici, per lo studio di questa malattia (recensito altrove 1,2). Di questi modelli, i più popolari sono indotta da ipossia (Hx) PAH nel topo e la Monocrotalina (MCT) e modelli SU5416-ipossia (SuHx) nel ratto. Nel modello Hx mouse, i topi sono esposti a 4 settimane di ipossia (sia normobariche o ipobarica, corrispondente ad un&#39;altitudine di 18.000 piedi con una FiO2 0,10), con lo sviluppo risultante proliferazione mediale, maggiore RV systpressioni Olic e lo sviluppo di ipertrofia ventricolare 3. MCT ad una singola dose di 60 mg / kg risultati in lesioni alle cellule endoteliali polmonari attraverso un meccanismo chiaro che poi si traduce nello sviluppo di PAH 4. SU5416 è un inibitore delle vascolari recettori del fattore di crescita endoteliale (VEGFR) 1 e 2 bloccante, e il trattamento con una singola iniezione sottocutanea di 60 mg / kg seguita da esposizione a ipossia cronica per 3 settimane risultati in ipertensione polmonare permanente con alterazioni patologiche simili a quello visto nella malattia umana, con la formazione di lesioni vascolari obliterante 5. Negli ultimi anni, sono stati sviluppati diversi modelli di topi transgenici per l&#39;ipertensione polmonare. Questi includono eliminazione diretta e le mutazioni del recettore proteina morfogenetica dell&#39;osso 2 (BMPR2), come BMPR2 mutazioni del gene sono presenti in entrambe le forme familiari e idiopatiche di PAH, eme ossigenasi-1 KO e IL-6 sovraespressione (recensione altrove 1,2). Questi diversi modelli di roditori di PH hanno diversi livelli di ipertensione polmonare, ipertrofia ventricolare e insufficienza RV. Mentre i vari modelli di topi transgenici ipossia e si traducono in PAH molto più mite rispetto al modello di ratto o 1, lo fa consentire la sperimentazione di diverse mutazioni genetiche e dei loro percorsi di segnalazione associate molecolari. Il modello MCT non causare gravi PAH, anche se MCT sembra essere tossico per le cellule endoteliali in diversi tessuti 4. Il modello SuHx è caratterizzata da modificazioni vascolari più simile a quella vista in PAH idiopatica negli esseri umani, anche se richiede sia esposizione manipolazione e ipossia farmacologica. Inoltre, in tutti questi modelli, ci può essere una disconnessione tra le variazioni istopatologiche, pressione polmonare e funzione ventricolare associati allo sviluppo di PAH. Questo è in contrasto con la malattia umana, in cui vi è solitamente un rapporto proporzionale tra cambiamenti istopatologici, la gravità della pulmonipertensione ary e il grado di insufficienza RV. Quindi, è necessario una caratterizzazione completa di questi modelli di roditori di PH, e implica valutazioni di funzione ventricolare destra (tipicamente mediante ecocardiografia), l&#39;emodinamica (di cateterismo cardiaco) e istopatologia del cuore e dei polmoni (dalla raccolta dei tessuti). In questo protocollo, si descrivono le tecniche di base utilizzate per la caratterizzazione di modelli emodinamica PAH nel ratto e nel topo. Queste tecniche generali possono essere applicati a qualsiasi studio del ventricolo destro e vasi polmonari e non è limitata a modelli di PAH. Visualizzare il RV mediante ecocardiografia è relativamente semplice nei ratti, ma è più impegnativo nei topi causa delle loro dimensioni e la geometria complessa del RV. Inoltre, alcuni surrogati utilizzati per la funzione RV quantificare, come TAPSE, arteria polmonare (PA) tempo di accelerazione e PA Doppler forma d&#39;onda dentellatura, non sono ben convalidati negli esseri umani e correlano solo debolmente con la valutazione di puipertensione lmonary e funzione ventricolare destra da emodinamica invasiva. Determinazione delle emodinamica RV è meglio farlo con un chiuso del torace, per mantenere gli effetti di una pressione negativa intratoracica con l&#39;ispirazione, anche se aperto cateterizzazione petto con un catetere impedenza per il calcolo di pressione-volume (PV) loop e una caratterizzazione emodinamico più dettagliata . Come con qualsiasi procedura, sviluppando l&#39;esperienza con le procedure è fondamentale per il successo sperimentale.

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		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Vevo 2100 Imaging System (120V)&#160;</td>
			<td style="width:183px;">VisualSonics, inc.&#160;</td>
			<td style="width:191px;">VS-11945</td>
			<td style="width:167px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Vevo 2100 Imaging Station&#160;</td>
			<td style="width:183px;">VisualSonics, inc.&#160;</td>
			<td style="width:191px;"></td>
			<td></td>
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		<tr height="27">
			<td height="27" style="height:27px;width:443px;">High-frequency Mechanical Transducers</td>
			<td style="width:183px;">VisualSonics, inc.&#160;</td>
			<td style="width:191px;">MS250, MS550D, MS400</td>
			<td></td>
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		<tr height="25">
			<td height="25" style="height:25px;width:443px;">Ultrasound Gel Parker&#160;</td>
			<td style="width:183px;">Laboratories Inc.&#160;</td>
			<td style="width:191px;">01-08</td>
			<td></td>
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			<td height="21" style="height:21px;">PowerLab 4/35</td>
			<td>ADInstruments</td>
			<td>ML765</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Labchart 8</td>
			<td>ADInstruments</td>
			<td></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">BP transducer with stopcock and cable</td>
			<td>ADInstruments</td>
			<td>MLT1199</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:443px;">BP transducer calibration kit</td>
			<td style="width:183px;">ADInstruments</td>
			<td style="width:191px;">MLA1052</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Mikro-Tip Pressure Catheter for mouse</td>
			<td>Millar</td>
			<td>SPR-1000</td>
			<td>Alternative catheter available from Scisense FT111B (mouse) and FT211B (rat)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Mikro-Tip Pressure Catheter for rat</td>
			<td style="width:183px;">Millar</td>
			<td style="width:191px;">SPR-513</td>
			<td>Alternative catheter available from Scisense FT111B (mouse) and FT211B (rat)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Millar Mikro-Tip&#160;ultra-miniature PV loop catheter for mice</td>
			<td>Millar</td>
			<td>PVR-1035</td>
			<td>Alternative catheter available from Scisense FT112 (mouse)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Millar Mikro-Tip ultra miniature PV loop catheter for rats</td>
			<td style="width:183px;">Millar</td>
			<td style="width:191px;">SPR-869</td>
			<td>Alternative catheter available from Scisense FT112 (mouse)</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Millar PV system MPVS-300&#160;</td>
			<td>Millar</td>
			<td>MPVS-300</td>
			<td></td>
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			<td height="21" style="height:21px;">4-0 Silk Black Braid 100 Yard Spool</td>
			<td>Roboz Surgical</td>
			<td>SUT-15-2</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">6-0 Silk Black Braid 100 Yard Spool</td>
			<td>Roboz Surgical</td>
			<td>SUT-14-1</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Iris Scissors, Delicate, Integra Miltex</td>
			<td>VWR</td>
			<td>21909-248</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">VWR Dissecting Scissors, Sharp/Blunt Tip</td>
			<td style="width:183px;">VWR</td>
			<td style="width:191px;">82027-588</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:443px;">VWR Delicate Scissors, 4 1/2&#34;</td>
			<td style="width:183px;">VWR</td>
			<td style="width:191px;">82027-582</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Two star Hemostats, Excelta</td>
			<td style="width:183px;">VWR</td>
			<td style="width:191px;">63042-090</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Neutral-buffered formalin</td>
			<td style="width:183px;">VWR</td>
			<td style="width:191px;">89370-094</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Crotaline</td>
			<td style="width:183px;">Sigma</td>
			<td style="width:191px;">C2401</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:443px;">SU5416</td>
			<td style="width:183px;">Tocris Biosciences</td>
			<td align="right" style="width:191px;">3037</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3.5X-45X Boom Stand Trinocular Zoom Stereo Microscope&#160;</td>
			<td>AmScope</td>
			<td>SM-3BX</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">PE (Polyethylene Tubing)-10</td>
			<td>Braintree Scientific Inc</td>
			<td style="width:191px;">PE10 36 FT</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PE (Polyethylene Tubing)-50</td>
			<td>Braintree Scientific Inc</td>
			<td style="width:191px;">PE50 36 FT</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PE (Polyethylene Tubing)-60</td>
			<td style="width:183px;">Braintree Scientific Inc</td>
			<td style="width:191px;">PE60 36 FT</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Tabletop Isoflurane Anesthesia Unit</td>
			<td>Kent Scientific</td>
			<td style="width:191px;">ACV-1205S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgisuite multi-functional surgical platform</td>
			<td>Kent Scientific</td>
			<td style="width:191px;">Surgisuite</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Retractor set</td>
			<td>Kent Scientific</td>
			<td>SURGI-5002</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Anesthesia induction chamber</td>
			<td>VetEquip</td>
			<td align="right" style="width:191px;">941443</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Anesthesia Gas filter canister</td>
			<td style="width:183px;">Kent Scientific</td>
			<td style="width:191px;">ACV-2001</td>
			<td></td>
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			<td height="21" style="height:21px;width:443px;">Rodent nose cone</td>
			<td style="width:183px;">VetEquip</td>
			<td align="right" style="width:191px;">921431</td>
			<td></td>
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hemodynamic characterization of rodent models of pulmonary arterial hypertension

Pulmonary arterial hypertension (PAH) is a rare disease of the pulmonary vasculature characterized by endothelial cell apoptosis, smooth muscle proliferation and obliteration of pulmonary arterioles. This in turn results in right ventricular (RV) failure, with significant morbidity and mortality. Rodent models of PAH, in the mouse and the rat, are important for understanding the pathophysiology underlying this rare disease. Notably, different models of PAH may be associated with different degrees of pulmonary hypertension, RV hypertrophy and RV failure. Therefore, a complete hemodynamic characterization of mice and rats with PAH is critical in determining the effects of drugs or genetic modifications on the disease. 
Here we demonstrate standard procedures for assessment of right ventricular function and hemodynamics in both rat and mouse PAH models. Echocardiography is useful in determining RV function in rats, although obtaining standard views of the right ventricle is challenging in the awake mouse. Access for right heart catheterization is obtained by the internal jugular vein in closed-chest mice and rats. Pressures can be measured using polyethylene tubing with a fluid pressure transducer or a miniature micromanometer pressure catheter. Pressure-volume loop analysis can be performed in the open chest. After obtaining hemodynamics, the rodent is euthanized. The heart can be dissected to separate the RV free wall from the left ventricle (LV) and septum, allowing an assessment of RV hypertrophy using the Fulton index (RV/(LV+S)). Then samples can be harvested from the heart, lungs and other tissues as needed.

Come giusto cateterizzazione cardiaca nei roditori è in genere una procedura terminale che non è applicabile a follow-up longitudinale, l&#39;ecocardiografia è un&#39;ottima alternativa non invasiva per lo screening e il follow-up 12. Mentre polmonare sistolica pressione arteriosa in PAH umana sulla ecocardiografia è di solito derivato da rigurgito della tricuspide che di solito è semplice da ottenere nella vista apicale, una tale visione non è affidabile ottenuto in roditori, impedendo la stima della p...

Watch this Scientific Journal Video about Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension at JoVE.com

Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension

Pulmonary arterial hypertension (PAH) is a disease of pulmonary arterioles that leads to their obliteration and the development of right ventricular failure. Rodent models of PAH are critical in understanding the pathophysiology of PAH. Here we demonstrate hemodynamic characterization, with right heart catheterization and echocardiography, in the mouse and rat.

Emodinamica Caratterizzazione di roditori modelli di ipertensione arteriosa polmonare

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

Research

JoVE Journal

Medicine

Magnetic Resonance Imaging Quantification of Pulmonary Perfusion using Calibrated Arterial Spin Labeling

Magnetic Resonance Imaging Quantificazione di perfusione polmonare mediante calibrati Etichettatura arteriosa Spin

This demonstrates a MR imaging method to measure the spatial distribution of pulmonary blood flow in healthy subjects 
during normoxia (inspired O2, ...

Ricerca

Medicina

Increasing Pulmonary Artery Pulsatile Flow Improves Hypoxic Pulmonary Hypertension in Piglets

Aumentare polmonare Arteria flusso pulsatile Migliora ipossica Ipertensione polmonare nei suinetti

Pulmonary arterial hypertension (PAH) is a disease affecting distal pulmonary arteries (PA). These arteries are deformed, leading to right ventricular ...

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

Tromboembolica cronica Ipertensione polmonare e valutazione dei ventricolare destra Funzione nel Maialino

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

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

Chirurgia Shunt, Destra cuore cateterizzazione, e Vascolare Morfometria in un modello di ratto per Flow-indotta ipertensione arteriosa polmonare

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 ...

Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats

Emodinamico invasivo caratterizzazione della sindrome Portal-ipertesi in ratti cirrotici

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. ...

Invasive Hemodynamic Monitoring of Aortic and Pulmonary Artery Hemodynamics in a Large Animal Model of ARDS

Monitoraggio emodinamico invasivo dell'emodinamica dell'arteria aortica e polmonare in un modello animale grande di ARDS

One of the leading causes of morbidity and mortality in patients with heart failure is right ventricular (RV) dysfunction, especially if it is due to ...

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

Induzione e fenotipizzazione dell'insufficienza cardiaca destra acuta in un modello animale di ipertensione polmonare tromboembolica cronica

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

Stenosi di sinistra atriale indotta arterializzazione venosa polmonare e l'ipertensione polmonare gruppo 2 nel ratto

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

The Left Pneumonectomy Combined with Monocrotaline or Sugen as a Model of Pulmonary Hypertension in Rats

La pneumonectomia sinistra combinato con Monocrotaline o Sugen come un modello di ipertensione polmonare in ratti

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats ...

Induction and Characterization of Pulmonary Hypertension in Mice using the Hypoxia/SU5416 Model

Induzione e caratterizzazione dell'ipertensione polmonare nei topi utilizzando il modello Ipossia/SU5416

Pulmonary Hypertension (PH) is a pathophysiological condition, defined by a mean pulmonary arterial pressure exceeding 25 mm Hg at rest, as assessed by ...