Name: evaluation of coronary flow reserve after myocardial ischemia reperfusion in rats
Uploaded: 2019-06-28T13:20:00
Description: Watch this Scientific Journal Video about Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats Video at JoVE.com

We would like to thank the Helmsley foundation for providing ultrasound equipment to perform our experiments. This work was supported by NIA R01 053585 grant.

All procedures were performed in accordance with protocols approved by the University of Louisville Institutional Animal Care and Use Committee (IACUC-approved protocol 18223) and the NIH Guide for the Care and Use of Laboratory Animals11.
1. Animals
<ol>
	<li>Use 4-month-old female Fisher 344 rats (BW~150-180 g) for the study.</li>
</ol>
2. Ultrasound Imaging before IR surgery
<ol>
	<li>Anesthetize the rat with isoflurane - induction chamber...

<ol>
	<li>Eltzschig, H. K., Eckle, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ischemia+and+reperfusion--from+mechanism+to+translation.">Ischemia and reperfusion--from mechanism to translation.</a> Nature medicine. 17 (11), 1391-1401 (2011).</li><li>French, C. J., Zaman, A. K., Sobel, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Failure+of+erythropoietin+to+render+jeopardized+ischemic+myocardium+amenable+to+incremental+salvage+by+early+reperfusion.">Failure of erythropoietin to render jeopardized ischemic myocardium amenable to incremental salvage by early reperfusion.</a> Coronary Artery Disease. 20 (4), 295-299 (2009).</li><li>Marzilli, M., Mariani, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ischemia-reperfusion+and+microvascular+dysfunction:+implications+for+salvage+of+jeopardized+myocardium+and+reduction+of+infarct+size.">Ischemia-reperfusion and microvascular dysfunction: implications for salvage of jeopardized myocardium and reduction of infarct size.</a> Italian Heart Journal. 2 Suppl 3, (2001).</li><li>Prasad, A., Gersh, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+microvascular+dysfunction+and+reperfusion+injury.">Management of microvascular dysfunction and reperfusion injury.</a> Heart (British Cardiac Society). 91 (12), 1530-1532 (2005).</li><li>Ishihara, M., 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=Impaired+coronary+flow+reserve+immediately+after+coronary+angioplasty+in+patients+with+acute+myocardial+infarction.">Impaired coronary flow reserve immediately after coronary angioplasty in patients with acute myocardial infarction.</a> British heart journal. 69 (4), 288-292 (1993).</li><li>Camici, P. G., d&#39;Amati, G., Rimoldi, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coronary+microvascular+dysfunction:+mechanisms+and+functional+assessment.">Coronary microvascular dysfunction: mechanisms and functional assessment.</a> Nature Reviews Cardiology. 12 (1), 48-62 (2015).</li><li>Chung, K. S., Nguyen, P. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+measures+of+coronary+microcirculation:+Taking+the+long+road+to+the+clinic.">Non-invasive measures of coronary microcirculation: Taking the long road to the clinic.</a> Journal of Nuclear Cardiology. , (2017).</li><li>Kelm, N. Q., 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=Adipose-derived+cells+improve+left+ventricular+diastolic+function+and+increase+microvascular+perfusion+in+advanced+age.">Adipose-derived cells improve left ventricular diastolic function and increase microvascular perfusion in advanced age.</a> PLoS One. 13 (8), e0202934 (2018).</li><li>Pan, M., 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=Late+recovery+of+coronary+flow+reserve+in+patients+successfully+treated+with+a+percutaneous+procedure.">Late recovery of coronary flow reserve in patients successfully treated with a percutaneous procedure.</a> Revista Espa&#241;ola de Cardiolog&#237;a. 56 (5), 459-464 (2003).</li><li>Samim, A., Nugent, L., Mehta, P. K., Shufelt, C., Bairey Merz, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+angina+and+microvascular+coronary+dysfunction.">Treatment of angina and microvascular coronary dysfunction.</a> Current treatment options in cardiovascular medicine. 12 (4), 355-364 (2010).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide for the Care and Use of Laboratory Animals. , (2011).</li><li>Ciuffreda, M. C., 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=Rat+experimental+model+of+myocardial+ischemia/reperfusion+injury:+an+ethical+approach+to+set+up+the+analgesic+management+of+acute+post-surgical+pain.">Rat experimental model of myocardial ischemia/reperfusion injury: an ethical approach to set up the analgesic management of acute post-surgical pain.</a> PloS one. 9 (4), (2014).</li><li>van de Hoef, T. P., Siebes, M., Spaan, J. A. E., Piek, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fundamentals+in+clinical+coronary+physiology:+why+coronary+flow+is+more+important+than+coronary+pressure.">Fundamentals in clinical coronary physiology: why coronary flow is more important than coronary pressure.</a> European Heart Journal. 36 (47), 3312-3319 (2015).</li><li>Naya, M., 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=Preserved+coronary+flow+reserve+effectively+excludes+high-risk+coronary+artery+disease+on+angiography.">Preserved coronary flow reserve effectively excludes high-risk coronary artery disease on angiography.</a> Journal of nuclear medicine : official publication, Society of Nuclear Medicine. 55 (2), 248-255 (2014).</li><li>Cortigiani, L., 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=Diagnostic+and+prognostic+value+of+Doppler+echocardiographic+coronary+flow+reserve+in+the+left+anterior+descending+artery.">Diagnostic and prognostic value of Doppler echocardiographic coronary flow reserve in the left anterior descending artery.</a> Heart (British Cardiac Society). 97 (21), 1758 (2011).</li><li>Kawata, T., 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=Prognostic+value+of+coronary+flow+reserve+assessed+by+transthoracic+Doppler+echocardiography+on+long-term+outcome+in+asymptomatic+patients+with+type+2+diabetes+without+overt+coronary+artery+disease.">Prognostic value of coronary flow reserve assessed by transthoracic Doppler echocardiography on long-term outcome in asymptomatic patients with type 2 diabetes without overt coronary artery disease.</a> Cardiovascular diabetology. 12, 121-121 (2013).</li></ol>

The major findings from the present study are that IR increases the resting LAD coronary artery velocity and impairs CFR, even in the absence of any residual angiographic stenosis.
Understanding the coronary physiology is an essential part of the clinical decision-making for cardiologists to treat coronary artery disease. CFR is one of the important functional parameters in understanding the pathophysiology of coronary microcirculation7,13</...

Myocardial ischemia reperfusion (IR) is a condition where blood supply is restricted to the heart followed by the restoration of perfusion and simultaneous reoxygenation1. Occlusion of coronary arteries can be caused by an embolus or cholesterol plaque rupture, which results in a severe imbalance of metabolic supply and demand, causing tissue hypoxia. Salvage of jeopardized myocardium, improve left ventricular function, and enhance survival in patients with acute myocardial infarction have been observed by the reperfusion therapy. However, after recanalization of the coronary artery, functional abnormalities of small coronary vessels may occur2,3,4,5. A significant proportion of patients, perhaps as many as 40%, do not regain microvascular and myocardial perfusion despite the restoration of coronary flow. Visualization and evaluation of the coronary microvasculature can be difficult in vivo, but there are a number of invasive and noninvasive techniques that can be used to assess parameters directly depending on the coronary microvascular function6,7. Also, the endothelial function can be assessed at the level of the coronary circulation via the CFR5.
Transthoracic Doppler echocardiography is a noninvasive tool which allows us to study coronary artery flow velocity and CFR5. CFR represents the ratio of maximal coronary blood flow to the resting coronary blood flow8. During the stress challenge, a normal heart increases coronary blood flow up to four times above the resting value. Cardiovascular risk increases when CFR is diminished9. Ishihara et al showed that the CFR was severely impaired immediately after the coronary angioplasty5. In the absence of coronary artery stenosis, CFR decreases during the coronary microvascular dysfunction and is present in about half of the patients with stable coronary artery disease10.
The overall goal of this method is noninvasive visualization of left anterior descending coronary artery (LAD) function in rats via echocardiography, which may be used to calculate CFR. This offers an important assessment tool for diagnosing microvascular dysfunction and evaluating potential therapeutic treatments.

<table>
	<tbody>
		<tr>
			<td>10 mL syringe</td>
			<td>BD Syringe</td>
			<td>302995</td>
			<td></td>
		</tr>
		<tr>
			<td>250S 13&#8211;24 MHz linear probe</td>
			<td>FUJIFILM VisualSonics Inc</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dobutamine hydrochloride</td>
			<td>Sigma</td>
			<td>D0676-10mg</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RRC</td>
			<td>27376</td>
			<td></td>
		</tr>
		<tr>
			<td>Legato 100 Syringe pump</td>
			<td>KD Scientific</td>
			<td>788100</td>
			<td></td>
		</tr>
		<tr>
			<td>Vevo 3100</td>
			<td>FUJIFILM VisualSonics Inc</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Winged infusion set, 27G x 1/2&#34;,</td>
			<td>Medline.com</td>
			<td>TMOSV27ELZ</td>
			<td></td>
		</tr>
	</tbody>
</table>

evaluation of coronary flow reserve after myocardial ischemia reperfusion in rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

For this study, we used 12 female Fisher 344 rats. We performed a stress test with Dobutamine and measured LAD coronary artery velocity before and 72 hours after the IR surgery. Before the IR surgery, resting LAD coronary artery velocity was measured as 423 &#177; 59 mm/s, which was increased after Dobutamine infusion (1005 &#177; 77mm/s) (Figure 3A). After 72 h of ischemia reperfusion, resting LAD coronary artery velocity was significantly higher compared to...

Watch this Scientific Journal Video about Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats Video at JoVE.com

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats Video (Video) | JoVE

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

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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