Name: ultrasound based assessment of coronary artery flow and coronary flow reserve using the pressure overload model in mice
Uploaded: 2015-04-13T13:00:00
Description: Watch this Scientific Journal Video about Ultrasound Based Assessment of Coronary Artery Flow and Coronary Flow Reserve Using the Pressure Overload Model in Mice at JoVE.com

We thank Fred Roberts for exemplary technical support and also appreciate the help from the histology core in Beth Israel Hospital. We thank Brigham Women&#8217;s Hospital Cardiovascular Physiology Core for providing with the instrumentation and the funds for this work. This work was supported in part by a Department of Medicine Sundry Fund.

OPMERKING: Alle procedures werden uitgevoerd bij muizen in overeenstemming met American Veterinary Medical Association (AVMA) richtlijnen en goedgekeurd Institutional Animal Care en gebruik Comités (IACUC) protocollen. 1. Studie Ontwerp <ol><li> Gebruik 8-10 weken oude mannelijke C57BL / 6 muizen (BW ~ 25 g) in de studie. </li><li> Willekeurige de muizen (n = 11) in twee groepen, de studiegroep geselecteerd aorta banding (n = 8) en de controlegroep (n = 3) sham geopereerd via thoracotomie...

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In deze ultrageluid gebaseerd onderzoek, werd niet-invasieve evaluatie van coronaire stroming reproduceerbaar onmiddellijk plaats, over dagen, in levende proefmuizen; voorts het protocol aangetoond potentieel kransslagader stoornis die in een vroeg stadium was en is geassocieerd met deficiëntie myocardiale perfusie detecteren. Deze methode zou uiteindelijk kunnen worden ingezet als klinisch hulpmiddel voor cardiovasculaire risicostratificatie en / of beoordelen van respons op therapeutische interventie. <p class="j...

Klinische aortastenose (AS) is algemeen bekend om een ​​progressieve toename in linker ventrikel (LV) afterload promoten. Om dit te compenseren chronisch stijgende hemodynamische belasting, LV hypertrofie (LVH) ontstaat als een adaptieve respons 1,2. De ontwikkeling van LVH wordt vaak geassocieerd met abnormaliteiten in coronaire microcirculatie. Men denkt dat microvasculaire disfunctie bijdraagt ​​chronische ischemie bij deze patiënten 5. Naast coronaire stroom 3,4, coronaire stroming reserve (CFR) vertegenwoordigt functionele wijziging van kransslagaders 1,3 en wordt gedefinieerd als de verhouding van de maximale stroomsnelheid in hyperemie uitgangswaarde stroomsnelheid of resting stroomsnelheid 4,6,7. CFR Verlagen van remodeling 1-3,5-9 en wordt gebruikt als een index van de mate van functionele ernst van coronaire disfunctie 1,10,17. Het is bekend verminderen bij vele vormen van gedilateerde cardiomyopathie 10 alsook coronaire stenosis 6. CFR is ook een prognostische marker voor slechte klinische resultaten 12. LV remodeling in de setting van cardiale disfunctie zoals ischemie of LVH gaat tevens gepaard met uitgebreide fibrose, veranderingen in coronaire microcirculatie en verdikking van de kransslagaders 1,2. Als gevolg van deze veranderingen in coronaire fysiologie, er waarschijnlijk remodelleren van de kransslagaders. Dit helpt de effecten van lage diffusie zuurstof en LV diastolische dysfunctie die kunnen leiden tot vatbaarheid voor myocardischemie 1,2,13 verzachten. Genetisch gemodificeerde muizen zijn nu een grote schaal voorkomt onderzoek tool voor het nabootsen van menselijke omstandigheden ziekten zoals coronaire atherosclerose 5,7,10,12,17. Vooral de druk overbelasting model bij muizen is op grote schaal bestudeerd 14,17. De trans-aorta vernauwing model (TAC) blijkt geassocieerd met uitgebreide fibrose en Coronary stenose resulteert gedeeltelijk uit mediale verdikking van kransslagaders en bijbehorende veranderingen in coronaire stromingspatronen 1,11,17,19 vergelijkbaar met wat wordt gezien in de omgeving van LVH bij mensen. Hoewel bekend is dat langdurige drukoverbelasting leidt tot hartdecompensatie in ongeveer 4-8 weken, de effecten op coronaire miek en stroomreserve in deze modellen, vroeg stadium van de ziekte, en in verschillende stadia na banding, nog duidelijk worden afgebakend. Talrijke stammen van muizen zijn momenteel beschikbaar voor onderzoeksdoeleinden, met inbegrip van goed gekarakteriseerde LDLR - / - of ApoE - / - muizen 10-12, en deze zijn ontwikkeling van gevoelige technieken gevraagd voor de beoordeling van de cardiovasculaire functie en morfologie in levende muizen 11-15. Dergelijke technieken omvatten MRI, PET, contrast CT hoogfrequente ultrasone en elektronenbundel tomografie 2,9,17,19, die alle voorzien veelbelovende alternatieven voor invasievemethoden zoals cardiale catheterisatie en coronaire angiografie 12. In muizen met zeer kleine afmetingen van de coronaire slagaders en hoge hartslag (HR), beeldvorming van coronaire circulatie vormt nog een technische uitdaging voor vele momenteel beschikbare technieken 4,12. Interessant is er een exponentiële toename van technische ontwikkelingen op het gebied van transthoracale Doppler echocardiografie (TTDE), waaronder de ontwikkeling van hoogfrequente matrix scan hoofden met centrale frequenties van 15 tot 50 MHz waardoor axiale resoluties van ongeveer 30-100 urn is, op een diepte van 8-40 mm, en frame rates van meer dan 400 frames gevangen / sec. Op hun beurt zijn TTDE-gebaseerde technieken ontwikkeld tot een potentieel krachtig instrument voor groter 2 of zelfs kleinere vaartuigen afbeelden zoals kransslagaders 5,12. Een andere kritische voorhand dat heeft toegestaan ​​onderzoekers aan diagnostische beeldvorming studies van het vaatstelsel te voeren in kleine eennimals is het zorgvuldig gecontroleerde gebruik van anesthetica die het hart en de ademhaling van de dieren tijdens het afbeelden 11 te behouden. Gecontroleerde anesthesie onderhoud is bijzonder belangrijk voor studies in verband met vasodilatatie bij muizen, en het effect van anesthesie moet ook verder worden onderzocht in de context 10,11. Bij mensen daarentegen, TTDE afgeleide CFR metingen een meer gebruikelijke hulpmiddel voor de beoordeling van vernauwde en niet belemmerd epicardiale coronaire slagaders, overwegend linker voorste dalende (LAD) coronaire arterie 5,16. Echter, de prognostische rol van CFR en coronaire stroming veranderingen in asymptomatische patiënten en muizen met een behouden LV systolische functie in rust minder onderzocht 16. Daarom was het doel van de studie was om eerst een duidelijke stap-voor-stap protocol vast, veranderingen in coronaire stroom met TTDE een drukoverbelasting muismodel evalueren; tweede, deze studie onderzocht de prognostische tekenificance van CFR en coronaire stroom veranderingen in reactie op overbelasting van stress druk in deze muizen. Onze hypothese was dat TTDE gebaseerde beoordeling van CFR en coronaire doorstroming nuttig in de vroege opsporing van coronaire dysfunctie dat LV dysfunctie kan voorafgaan kunnen zijn.

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ultrasound based assessment of coronary artery flow and coronary flow reserve using the pressure overload model in mice

Transthoracic Doppler echocardiography (TTDE) is a clinically useful, noninvasive tool for studying coronary artery flow velocity and coronary flow reserve (CFR) in humans. Reduced CFR is accompanied by marked intramyocardial and pericoronary fibrosis and is used as an indication of the severity of dysfunction. This study explores, step-by-step, the real-time changes measured in the coronary flow velocity, CFR and systolic to diastolic peak velocity (S/D) ratio in the setting of an aortic banding model in mice. By using a Doppler transthoracic imaging technique that yields reproducible and reliable data, the method assesses changes in flow in the septal coronary artery (SCA), for a period of over two weeks in mice, that previously either underwent aortic banding or thoracotomy. 
During imaging, hyperemia in all mice was induced by isoflurane, an anesthetic that increased coronary flow velocity when compared with resting flow. All images were acquired by a single imager. Two ratios, (1) CFR, the ratio between hyperemic and baseline flow velocities, and (2) systolic (S) to diastolic (D) flow were determined, using a proprietary software and by two independent observers. Importantly, the observed changes in coronary flow preceded LV dysfunction as evidenced by normal LV mass and fractional shortening (FS). 
The method was benchmarked against the current gold standard of coronary assessment, histopathology. The latter technique showed clear pathologic changes in the coronary artery in the form of peri-coronary fibrosis that correlated to the flow changes as assessed by echocardiography. 
The study underscores the value of using a non-invasive technique to monitor coronary circulation in mouse hearts. The method minimizes redundant use of research animals and demonstrates that advanced ultrasound-based indices, such as CFR and S/D ratios, can serve as viable diagnostic tools in a variety of investigational protocols including drug studies and the study of genetically modified strains.

Van de 11 muizen die werden bestudeerd (banded, n = 8 en sham, n = 3) werden voldoende en reproduceerbare beelden verkregen door een enkele waarnemer op verschillende tijdstippen: bij aanvang (D-1), D2, D6 en D13 . Ook de stroomsnelheid bij de constrictieve plaats werd gemeten als 2225 ± 110,9 mm / s, in vergelijking met 277,5 ± 10,51 mm / s in de sham muizen op de dag na de operatie (p &lt;0,05). De toename van de snelheid was de verificatie van de succesvolle oprichting van de druk overbelasting model. De SCA stroom...

Watch this Scientific Journal Video about Ultrasound Based Assessment of Coronary Artery Flow and Coronary Flow Reserve Using the Pressure Overload Model in Mice at JoVE.com

Ultrasound Based Assessment of Coronary Artery Flow and Coronary Flow Reserve Using the Pressure Overload Model in Mice

Coronary flow reserve (CFR) is useful for assessment of myocardial oxygen demand and evaluation of cardiovascular risk. This study establishes a step-by-step transthoracic Doppler echocardiographic (TTDE) method for longitudinal monitoring of the changes in CFR, as measured from coronary artery in mice, under the experimental pressure overload of aortic banding.

Echografie gebaseerde beoordeling van coronaire Flow en Coronaire Flow Reserve Met behulp van de druk overbelasting Model in Muizen

Transthoracic Doppler echocardiography (TTDE) is a clinically useful, noninvasive tool for studying coronary artery flow velocity and coronary flow ...

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Echografie Beoordeling van endotheel-afhankelijke Flow-gemedieerde vaatverwijding van de arteria brachialis in Clinical Research

The vascular endothelium is a monolayer of cells that cover the interior of blood vessels and provide both structural and functional roles. The ...

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

Permanente Ligatie van de linker anterior Aflopend Kransslagader in Muizen: Een model van Post-myocardinfarct Remodelling en hartfalen

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 ...

Murine Left Anterior Descending (LAD) Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

Murine Left Anterior Descending LAD Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

Murine Left Anterior Descending (LAD) coronaire Ligatie: een verbeterde en vereenvoudigd model voor myocardinfarct

Ischemic heart disease (IHD), or acute coronary syndrome (ACS), is one of the leading causes of death in the United States. IHD is characterized by ...

Direct Re-implantation of Left Coronary Artery into the Aorta in Adults with Anomalous Origin of Left Coronary Artery from the Pulmonary Artery (ALCAPA)

Direct Re-implantation of Left Coronary Artery into the Aorta in Adults with Anomalous Origin of Left Coronary Artery from the Pulmonary Artery ALCAPA

Direct Re-implantatie van linker kransslagader in de aorta in Volwassenen met Abnormale Origin of linker kransslagader van de longslagader (ALCAPA)

Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital anomaly which is one of leading causes of myocardial ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

Isometrische Contractility meting van de muis mesenterische slagader met behulp van draad Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Evaluatie van coronaire stroom reserve na myocardiale ischemie reperfusie bij ratten

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

Murine Myocardial Infarction Model using Permanent Ligation of Left Anterior Descending Coronary Artery

Murine myocardinfarct model met behulp van permanente Ligatie van linker anterieure aflopende coronaire slagader

Myocardial infarction (MI) and acute coronary diseases are among the most prominent causes of death in population with western lifestyle. The murine ...