Name: cardiac response to &#946;-adrenergic stimulation determined by pressure-volume loop analysis
Uploaded: 2021-05-19T14:00:00
Description: Watch this Scientific Journal Video about Cardiac Response to ÃŽÂ²-Adrenergic Stimulation Determined by Pressure-Volume Loop Analysis at JoVE.com

我们感谢Manuela Ritzal，Hans-Peter Gensheimer，Christin Richter和海德堡大学Interfakultäre Biomedizinische Forschungseinrichtung（IBF）的团队提供专家技术援助。
这项工作得到了DZHK（德国心血管研究中心），BMBF（德国教育和研究部），巴登 - 符腾堡州联邦国家创新基金会和Deutsche Forschungsgemeinschaft（DFG，德国研究基金会）项目ID 239283807 - TRR 152，FOR 2289和协作研究中心（SFB）1118的支持。

所有动物实验均根据卡尔斯鲁厄地区委员会和海德堡大学的规定（AZ 35-9185.82/A-2/15，AZ 35-9185.82/A-18/15，AZ 35-9185.81/G131/15，AZ 35-9185.81/G121/17）的规定批准和进行，符合欧洲议会关于保护用于科学目的的动物的指令2010/63/ EU的指南。该协议中显示的数据来自野生型C57Bl6 / N雄性小鼠（17±1.4周龄）。在海德堡医学院的动物设施（IBF）中，小鼠在指定的无病原体条件下维持。将小鼠置于12小时的光暗循环中?...

<ol>
	<li>Bacmeister, 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=Inflammation+and+fibrosis+in+murine+models+of+heart+failure.">Inflammation and fibrosis in murine models of heart failure.</a> Basic Research in Cardiology. 114 (3), 19 (2019).</li><li>Hartupee, J., Mann, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurohormonal+activation+in+heart+failure+with+reduced+ejection+fraction.">Neurohormonal activation in heart failure with reduced ejection fraction.</a> Nature Reviews Cardiology. 14 (1), 30-38 (2017).</li><li>Hasenfuss, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+human+cardiovascular+disease,+heart+failure+and+hypertrophy.">Animal models of human cardiovascular disease, heart failure and hypertrophy.</a> Cardiovascular Research. 39 (1), 60-76 (1998).</li><li>Lefkowitz, R. J., Rockman, H. A., Koch, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catecholamines,+cardiac+beta-adrenergic+receptors,+and+heart+failure.">Catecholamines, cardiac beta-adrenergic receptors, and heart failure.</a> Circulation. 101 (14), 1634-1637 (2000).</li><li>Cingolani, O. H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pressure-volume+relation+analysis+of+mouse+ventricular+function.">Pressure-volume relation analysis of mouse ventricular function.</a> The American Journal of Physiology-Heart and Circulatory Physiology. 301, 2198-2206 (2011).</li><li>Bacmeister, 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=Assessment+of+PEEP-Ventilation+and+the+Time+Point+of+Parallel-Conductance+Determination+for+Pressure-Volume+Analysis+Under+beta-Adrenergic+Stimulation+in+Mice.">Assessment of PEEP-Ventilation and the Time Point of Parallel-Conductance Determination for Pressure-Volume Analysis Under beta-Adrenergic Stimulation in Mice.</a> Frontiers in Cardiovascular Medicine. 6, 36 (2019).</li><li>Segin, 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=Cardiomyocyte-Specific+Deletion+of+Orai1+Reveals+Its+Protective+Role+in+Angiotensin-II-Induced+Pathological+Cardiac+Remodeling.">Cardiomyocyte-Specific Deletion of Orai1 Reveals Its Protective Role in Angiotensin-II-Induced Pathological Cardiac Remodeling.</a> Cells. 9 (5), (2020).</li><li>Clark, J. E., Marber, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advancements+in+pressure-volume+catheter+technology+-+stress+remodelling+after+infarction.">Advancements in pressure-volume catheter technology - stress remodelling after infarction.</a> Experimental Physiology. 98 (3), 614-621 (2013).</li><li>Glower, D. D., 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=Linearity+of+the+Frank-Starling+relationship+in+the+intact+heart:+the+concept+of+preload+recruitable+stroke+work.">Linearity of the Frank-Starling relationship in the intact heart: the concept of preload recruitable stroke work.</a> Circulation. 71 (5), 994-1009 (1985).</li><li>Winter, E. 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=Left+ventricular+function+in+the+post-infarct+failing+mouse+heart+by+magnetic+resonance+imaging+and+conductance+catheter:+a+comparative+analysis.">Left ventricular function in the post-infarct failing mouse heart by magnetic resonance imaging and conductance catheter: a comparative analysis.</a> Acta Physiologica. 194 (2), 111-122 (2008).</li><li>Krenz, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conductance,+admittance,+and+hypertonic+saline:+should+we+take+ventricular+volume+measurements+with+a+grain+of+salt.">Conductance, admittance, and hypertonic saline: should we take ventricular volume measurements with a grain of salt.</a> Journal of Applied Physiology. 107 (6), 1683-1684 (2009).</li><li>Pacher, P., Nagayama, T., Mukhopadhyay, P., Batkai, S., Kass, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+cardiac+function+using+pressure-volume+conductance+catheter+technique+in+mice+and+rats.">Measurement of cardiac function using pressure-volume conductance catheter technique in mice and rats.</a> Nature Protocols. 3 (9), 1422-1434 (2008).</li><li>Wei, A. E., Maslov, M. Y., Pezone, M. J., Edelman, E. R., Lovich, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+pressure-volume+conductance+catheters+in+real-time+cardiovascular+experimentation.">Use of pressure-volume conductance catheters in real-time cardiovascular experimentation.</a> Heart, Lung and Circulation. 23 (11), 1059-1069 (2014).</li><li>van Hout, G. P., 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=Admittance-based+Pressure-Volume+Loops+versus+gold+standard+cardiac+magnetic+resonance+imaging+in+a+porcine+model+of+myocardial+infarction.">Admittance-based Pressure-Volume Loops versus gold standard cardiac magnetic resonance imaging in a porcine model of myocardial infarction.</a> Physiological Reports. 2 (4), 00287 (2014).</li><li>Wei, C. L., Shih, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calibration+Capacity+of+the+Conductance-to-Volume+Conversion+Equations+for+the+Mouse+Conductance+Catheter+Measurement+System.">Calibration Capacity of the Conductance-to-Volume Conversion Equations for the Mouse Conductance Catheter Measurement System.</a> IEEE Transactions on Biomedical Engineering. 56 (6), 1627-1634 (2009).</li><li>Das, S., MacDonald, K., Chang, H. Y., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+of+mouse+lung+intubation.">A simple method of mouse lung intubation.</a> Journal of Visualized Experiments. (73), e50318 (2013).</li><li>Faul, F., Erdfelder, E., Lang, A. G., Buchner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=G*Power+3:+a+flexible+statistical+power+analysis+program+for+the+social,+behavioral,+and+biomedical+sciences.">G*Power 3: a flexible statistical power analysis program for the social, behavioral, and biomedical sciences.</a> Behavior Research Methods. 39 (2), 175-191 (2007).</li><li>Weiss, J. L., Frederiksen, J. W., Weisfeldt, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemodynamic+determinants+of+the+time-course+of+fall+in+canine+left+ventricular+pressure.">Hemodynamic determinants of the time-course of fall in canine left ventricular pressure.</a> Journal of Clinical Investigation. 58 (3), 751-760 (1976).</li><li>Faul, F., Erdfelder, E., Lang, A. G., Buchner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=G*Power+3:+a+flexible+statistical+power+analysis+program+for+the+social,+behavioral,+and+biomedical+sciences.">G*Power 3: a flexible statistical power analysis program for the social, behavioral, and biomedical sciences.</a> Behavioral Research Methods. 39 (2), 175-191 (2007).</li><li>Jacoby, 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=Direct+comparison+of+magnetic+resonance+imaging+and+conductance+microcatheter+in+the+evaluation+of+left+ventricular+function+in+mice.">Direct comparison of magnetic resonance imaging and conductance microcatheter in the evaluation of left ventricular function in mice.</a> Basic Research in Cardiology. 101 (1), 87-95 (2006).</li><li>Georgakopoulos, D., Kass, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimation+of+parallel+conductance+by+dual-frequency+conductance+catheter+in+mice.">Estimation of parallel conductance by dual-frequency conductance catheter in mice.</a> The American Journal of Physiology-Heart and Circulatory Physiology. 279 (1), 443-450 (2000).</li><li>Calligaris, S. D., Ricca, M., Conget, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+stress+test+induced+by+dobutamine+and+monitored+by+cardiac+catheterization+in+mice.">Cardiac stress test induced by dobutamine and monitored by cardiac catheterization in mice.</a> Journal of Visualized Experiments. (72), e50050 (2013).</li><li>Abraham, D., Mao, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+Pressure-Volume+Loop+Analysis+Using+Conductance+Catheters+in+Mice.">Cardiac Pressure-Volume Loop Analysis Using Conductance Catheters in Mice.</a> Journal of Visualized Experiments. (103), e52942 (2015).</li><li>Pearce, J. A., Porterfield, J. E., Larson, E. R., Valvano, J. W., Feldman, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+considerations+in+catheter+based+estimation+of+left+ventricular+volume.">Accuracy considerations in catheter based estimation of left ventricular volume.</a> Conference proceedings - IEEE engineering in medicine and biology society. 2010, 3556-3558 (2010).</li><li>Nielsen, J. 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=Left+ventricular+volume+measurement+in+mice+by+conductance+catheter:+evaluation+and+optimization+of+calibration.">Left ventricular volume measurement in mice by conductance catheter: evaluation and optimization of calibration.</a> The American Journal of Physiology-Heart and Circulatory Physiology. 293 (1), 534-540 (2007).</li><li>Townsend, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+Pressure+Volume+Loops+in+the+Mouse.">Measuring Pressure Volume Loops in the Mouse.</a> Journal of Visualized Experiments. (111), e53810 (2016).</li><li>Barnabei, M. S., Palpant, N. J., Metzger, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+genetic+background+on+ex+vivo+and+in+vivo+cardiac+function+in+several+commonly+used+inbred+mouse+strains.">Influence of genetic background on ex vivo and in vivo cardiac function in several commonly used inbred mouse strains.</a> Physiological Genomics. 42 (2), 103-113 (2010).</li><li>Oosterlinck, W., Vanderper, A., Flameng, W., Herijgers, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucose+tolerance+and+left+ventricular+pressure-volume+relationships+in+frequently+used+mouse+strains.">Glucose tolerance and left ventricular pressure-volume relationships in frequently used mouse strains.</a> Journal of Biomedicine and Biotechnology. 2011, 281312 (2011).</li><li>Guo, X., Kono, Y., Mattrey, R., Kassab, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphometry+and+strain+distribution+of+the+C57BL/6+mouse+aorta.">Morphometry and strain distribution of the C57BL/6 mouse aorta.</a> The American Journal of Physiology-Heart and Circulatory Physiology. 283 (5), 1829-1837 (2002).</li><li>Weiss, R. M., Ohashi, M., Miller, J. D., Young, S. G., Heistad, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcific+aortic+valve+stenosis+in+old+hypercholesterolemic+mice.">Calcific aortic valve stenosis in old hypercholesterolemic mice.</a> Circulation. 114 (19), 2065-2069 (2006).</li></ol>

在这里，我们提供了一种方案来分析在增加β肾上腺素能刺激下的小鼠体内心脏功能。该程序可用于解决转基因小鼠或干预中心脏功能的基线参数和肾上腺素能储备（例如，正性肌力和变时性）。与其他确定心脏功能的方法相比，压力 - 体积环（PVL）测量最突出的优势是分析内在的，与负荷无关的心脏功能。所有其他方法（例如，MRI 和超声心动图）只能评估心脏功能的负荷依赖性参数，尤其是无?...

心血管疾病通常影响心脏功能。本期指出了动物疾病模型中评估体内详细心脏功能的重要性。动物实验被三个Rs（3R）指导原则（减少/改进/替换）的框架所包围。在当前发育水平上理解涉及全身反应（即心血管疾病）的复杂病理的情况下，主要选择是改进可用的方法。由于变异性较小，精炼还会导致所需动物数量的减少，从而提高分析和结论的力量。此外，心脏收缩力测量与心脏病的动物模型相结合，包括由神经体液刺激或主动脉带等压力过载引起的模型，其模拟例如改变的儿茶酚胺/β肾上腺素能水平1，2，3，4，为临床前研究提供了一种强有力的方法。考虑到基于导管的方法仍然是深入评估心脏收缩力5的最广泛使用的方法，我们旨在根据先前的经验（包括对该方法的具体参数的评估）在β肾上腺素能刺激期间通过压力 - 体积环（PVL）测量来精确测量小鼠体内心脏功能6， 7.
为了确定心脏血流动力学参数，可以使用包括成像或基于导管的技术的方法。这两种选择都伴随着优点和缺点，需要为各自的科学问题仔细考虑。成像方法包括超声心动图和磁共振成像（MRI）;两者都已成功用于小鼠。超声心动图测量涉及小鼠高心率所需的高速探头的高初始成本;这是一种相对简单的非侵入性方法，但在理想情况下应该具有识别和可视化心脏结构经验的操作员中，它是可变的。此外，不能直接进行压力测量，并且通过尺寸大小和流量测量的组合获得计算。另一方面，它的优点是可以对同一动物进行多次测量，并且可以监测心脏功能，例如在疾病进展期间。关于体积测量，MRI是金标准程序，但与超声心动图类似，无法进行直接压力测量，只能获得预载荷相关参数8。限制因素还包括可用性、分析工作量和运营成本。在这里，基于导管的测量心脏功能的方法是一个很好的替代方案，它还可以直接监测心内压力和确定与负荷无关的收缩性参数，如预负荷可招募卒中（PRSW）9。然而，通过压力 - 电导管（通过电导率测定）测量的心室体积小于MRI测量的心室体积，但组差异保持在相同的范围内10。为了确定可靠的体积值，需要进行相应的校准，这是PVL测量过程中的关键步骤。它将体积校准比色皿中血液电导率的离体测量（电导率到体积的转换）与体内分析相结合，以在推注高渗盐水11，12期间心肌平行电导。除此之外，导管在心室内的位置以及电极沿心室纵轴的正确方向对于检测它们产生的周围电场的能力至关重要。仍然随着小鼠心脏尺寸的减小，可以避免由导管脑室内方向变化产生的伪影，即使在扩张的心室5，10中，但伪影可以在β肾上腺素能刺激下进化6，13。除了电导方法之外，基于导纳的方法的发展似乎避免了校准步骤，但这里的体积值被高估了14，15。
由于小鼠是心血管研究中最重要的临床前模型之一，并且心脏的β-肾上腺素能储备是心脏生理学和病理学的核心兴趣，我们在这里提出了一个改进的方案，通过PVL测量在β肾上腺素能刺激期间通过PVL测量来确定小鼠体内的心脏功能。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1.4F SPR-839 catheter</td>
			<td>Millar Instruments, USA</td>
			<td>840-8111</td>
			<td></td>
		</tr>
		<tr>
			<td>1 ml syringes</td>
			<td>Beckton Dickinson, USA</td>
			<td>REF303172</td>
			<td></td>
		</tr>
		<tr>
			<td>Bio Amplifier</td>
			<td>ADInstruments, USA</td>
			<td>FE231</td>
			<td></td>
		</tr>
		<tr>
			<td>Bridge-Amplifier</td>
			<td>ADInstruments, USA</td>
			<td>FE221</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Roth, Germany</td>
			<td>8076.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine hydrochloride</td>
			<td>Bayer, Germany</td>
			<td>4007221026402</td>
			<td></td>
		</tr>
		<tr>
			<td>Calibration cuvette</td>
			<td>Millar, USA</td>
			<td>910-1049</td>
			<td></td>
		</tr>
		<tr>
			<td>Differential pressure transducer MPX</td>
			<td>Hugo Sachs Elektronik- Harvard Apparatus, Germany</td>
			<td>Type 39912</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont Forceps #5/45</td>
			<td>Fine Science tools Inc.</td>
			<td>11251-35</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont Forceps #7B</td>
			<td>Fine Science tools Inc.</td>
			<td>11270-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Graefe Forceps</td>
			<td>Fine Science tools Inc.</td>
			<td>11051-10</td>
			<td></td>
		</tr>
		<tr>
			<td>GraphPad Prism</td>
			<td>GraphPad Software</td>
			<td>Ver. 8.3.0</td>
			<td></td>
		</tr>
		<tr>
			<td>EcoLab-PE-Micotube</td>
			<td>Smiths, USA</td>
			<td>004/310/168-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Etomidate Lipuro</td>
			<td>Braun, Germany</td>
			<td>2064006</td>
			<td></td>
		</tr>
		<tr>
			<td>Excel</td>
			<td>Microsoft</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin</td>
			<td>Ratiopharm, Germany</td>
			<td>R26881</td>
			<td></td>
		</tr>
		<tr>
			<td>Hot plate and control unit</td>
			<td>Labotec, Germany</td>
			<td>Hot Plate 062</td>
			<td></td>
		</tr>
		<tr>
			<td>Isofluran</td>
			<td>Baxter, Germany</td>
			<td>HDG9623</td>
			<td></td>
		</tr>
		<tr>
			<td>Isofluran Vaporizer</td>
			<td>Abbot</td>
			<td>Vapor 19.3</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoprenalinhydrochloride</td>
			<td>Sigma-Aldrich, USA</td>
			<td>I5627</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine Bore Polythene tubing 0.61 mm OD, 0.28 mm ID</td>
			<td>Smiths Medical International Ltd, UK</td>
			<td>Ref. 800/100/100</td>
			<td></td>
		</tr>
		<tr>
			<td>MiniVent ventilator for mice</td>
			<td>Hugo Sachs Elektronik- Harvard Apparatus, Germany</td>
			<td>Type 845</td>
			<td></td>
		</tr>
		<tr>
			<td>MPVS Ultra PVL System</td>
			<td>Millar Instruments, USA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>AppliChem, Germany</td>
			<td>A3597</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl 0.9% isotonic</td>
			<td>Braun, Germany</td>
			<td>2350748</td>
			<td></td>
		</tr>
		<tr>
			<td>Pancuronium-bromide</td>
			<td>Sigma-Aldrich, USA</td>
			<td>BCBQ8230V</td>
			<td></td>
		</tr>
		<tr>
			<td>Perfusor 11 Plus</td>
			<td>Harvard Apparatus</td>
			<td>Nr. 70-2209</td>
			<td></td>
		</tr>
		<tr>
			<td>Powerlab 4/35 control unit</td>
			<td>ADInstruments, USA</td>
			<td>PL3504</td>
			<td></td>
		</tr>
		<tr>
			<td>Rechargeable cautery-Set</td>
			<td>Faromed, Germany</td>
			<td>09-605</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Fine Science tools Inc.</td>
			<td>140094-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Software LabChart 7 Pro</td>
			<td>ADInstruments, USA</td>
			<td>LabChart 7.3 Pro</td>
			<td></td>
		</tr>
		<tr>
			<td>Standard mouse food</td>
			<td>LASvendi GmbH, Germany</td>
			<td>Rod18</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereo microscope</td>
			<td>Zeiss, Germany</td>
			<td>Stemi 508</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical suture 8/0</td>
			<td>Suprama, Germany</td>
			<td>Ch.B.03120X</td>
			<td></td>
		</tr>
		<tr>
			<td>Venipuncture-cannula Venflon Pro Safty 20-gauge</td>
			<td>Beckton Dickinson, USA</td>
			<td>393224</td>
			<td></td>
		</tr>
		<tr>
			<td>Vessel Cannulation Forceps</td>
			<td>Fine Science tools Inc.</td>
			<td>00574-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Water bath</td>
			<td>Thermo Fisher Scientific, USA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe filter (Filtropur S 0.45)</td>
			<td>Sarstedt, Germany</td>
			<td>Ref. 83.1826</td>
			<td></td>
		</tr>
	</tbody>
</table>

cardiac response to &#946;-adrenergic stimulation determined by pressure-volume loop analysis

心脏功能的测定是心血管疾病动物模型中可靠的终点分析，以表征特定治疗对心脏的影响。由于遗传操作的可行性，小鼠已成为研究心脏功能和寻找新的潜在治疗靶点的最常见哺乳动物动物模型。在这里，我们描述了一种方案，用于在基础条件下使用压力 - 体积环测量和分析来确定体内心脏功能，并通过静脉输注增加浓度的异丙肾上腺素的β肾上腺素能刺激。我们提供完善的治疗方案，包括考虑到正呼气末压力的通气支持，以改善胸腔开胸测量期间的负面影响，以及强效镇痛（丁丙诺啡），以避免手术过程中疼痛引起的无法控制的心肌压力。总而言之，对程序的详细说明和对可能陷阱的讨论使高度标准化和可重复的压力 - 体积循环分析成为可能，通过防止可能的方法学偏倚来减少动物被排除在实验队列之外。

压力体积环（PVL）测量是分析药物心脏药效学和研究正常和病理条件下转基因小鼠模型心脏表型的强大工具。该协议允许评估成年小鼠模型中的心脏β肾上腺素能储备。在这里，我们描述了异氟醚麻醉联合丁丙诺啡（镇痛药）和泛库溴铵（肌肉松弛剂）下的开胸方法，其重点是通过股静脉导管输注异丙肾上腺素浓度来对β肾上腺素能刺激的心脏反应。该方案中显示的一些代表性数据来自野生型C57Bl6...

Watch this Scientific Journal Video about Cardiac Response to ÃŽÂ²-Adrenergic Stimulation Determined by Pressure-Volume Loop Analysis at JoVE.com

Cardiac Response to ÃƒÅ½Ã‚Â²-Adrenergic Stimulation Determined by Pressure-Volume Loop Analysis

在这里，我们描述了在增加静脉输注的异丙肾上腺素剂量下的心脏压力 - 体积环路分析，以确定小鼠的内在心脏功能和β肾上腺素能储备。我们使用改进的开胸方法进行压力 - 体积回路测量，其中我们包括具有正呼气末压的通气。

通过压力-体积环路分析确定心脏对β肾上腺素能刺激的反应

Determination of the cardiac function is a robust endpoint analysis in animal models of cardiovascular diseases in order to characterize effects of ...

我们的开胸方法，其中我们包括具有正呼气期末压的通气，对于评估和分析体内心脏功能以及β-肾上腺素能刺激的增加具有重要意义。该程序可用于解决与负荷无关的心脏功能的基线参数，以及转基因小鼠或干预中的β-肾上腺素能储备。在确认对踏板反射缺乏反应后，用70%乙醇消毒麻醉小鼠的左后肢，并做一个切口以暴露左股静脉。用烧灼术对上腹部动脉和静脉进行爆破。为了更好地看到股静脉，视野通过缝合线穿过腹部。然后，将缝合线放置在导管通道的远端，以固定股静脉。在股静脉下缝合线，并准备将颅骨结到穿刺部位。用连接到1毫升注射器的准备好的微管刺穿股静脉，并用打结的缝合线将管子固定在血管内。为了抵消任何液体损失，使用自动注射器泵以15微升的输注速率输注0.9%氯化钠，补充12.5%白蛋白，并用预热的0.9%氯化钠水合暴露的组织。要进入胸部，请用70%乙醇冲洗胸部，并在xiphoid过程下方切开皮肤。将胸肌与胸壁钝分开，并用镊子抬起胸壁，让胸壁被切开，直到隔膜完全可见。从下面切开横膈膜以暴露心脏顶点，并使用镊子小心地去除心包。在左侧进行有限的切除术，并在下腔静脉下方通过缝合线，以减轻下游的预紧力。使用25号套管，轻轻刺穿心尖，并用压力容积导管替换套管，直到所有电极都在心室内。然后，轻轻调整导管的位置，直到获得矩形环。要获得压力 - 体积回路测量值，请对感兴趣的心功能参数进行在线分析，并等待获得稳态心脏功能。将呼吸器停在呼气末位置，并记录基线参数。3至5秒后，使用镊子将缝合线抬起下腔静脉下方，以获得预负荷独立参数，然后打开呼吸机。等待至少30秒进行第二次闭塞，直到血流动力学参数稳定。在基础条件下获得测量结果后，将异丙肾上醇加载的注射器安装到注射器泵上。等待至少两分钟，直到观察到新的稳态心脏功能。在呼气末位置停止呼吸器之前，并记录基线参数。3至5秒后，在下腔静脉下方抬起缝合线以获得预紧力独立参数，并等待至少30秒进行第二次闭塞。然后安装含有下一个异丙肾上腺素浓度的注射器，并重复基线，并预加载独立参数记录。为了进行平行电导校准，在最后一次异丙肾上腺素剂量反应测量后，将装有15%氯化钠溶液的注射器连接到股导管，并小心地注入5微升的高渗溶液，直到压力 - 体积回路稍微向右移动。等到环路恢复到稳态，然后在呼气结束时停止呼吸器，并立即注射10微升15%氯化钠的推注。然后检查压力-体积回路是否已大大扩大，并向右移动。要进行电导与容积比，等待5分钟，直到高渗盐水推注完全稀释，然后再取出导管，并使用1毫升注射器，配有21号套管，从跳动心脏的左心室中除去至少600微升的血液。然后将血液转移到37摄氏度水浴中的预热校准比色皿中，具有已知体积的圆柱体，并将压力容积导管中央放入每个圆瓶中以记录电导率。使用这种开放式胸压容积环测量方法，如果导管正确放置在心室内，则整个心脏周期将由矩形压力容积环表示。如图所示，收缩期从等压收缩阶段开始，在此期间，两个心脏瓣膜都关闭。当心室压力超过主动脉压力时，主动脉瓣打开，血液在射血阶段被泵入主动脉。随后，当主动脉压超过心室压力时，主动脉瓣关闭，舒张开始。在等气压松弛期间，心室压力下降，直到心房压力超过心室压力，二尖瓣打开。被动舒张充盈，其特征在于舒张末期压力 - 容量关系，然后发生直到下一个心脏周期开始。由于它能够独立于前负荷确定心脏功能，因此压力 - 体积分析可以提供对心脏功能，心脏收缩力的详细见解。例如，在该分析中，异丙肾上腺素诱导了对每个测量的心脏功能和心脏收缩力参数的显着影响。相反，观察到舒张压参数的降低响应于异丙肾上腺素浓度的增加。在左心室内精确放置导管对于获得无伪影的可重复袢至关重要。为了优化导管放置，我们更喜欢开胸方法。在PV环路测量之后，可以确定异体异体性斑越，并且可以分离组织，以识别由治疗和/或遗传修饰引起的分子和组织病理学改变。该技术为在正常和病理条件下分析小鼠模型中的β-肾上腺素能储备铺平了道路，并且可以用作测试其他刺激的模板。

Research

Education

JoVE Core

JoVE Journal

Pharmacology

Medicine

Unit 1

Cardiac Response to &#946;-Adrenergic Stimulation Determined by Pressure-Volume Loop Analysis

Drugs Acting on Autonomic Nervous System: Adrenergic Agonists and Antagonists Agents

SCIENTISTS IN ACTION

A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

一个小鼠模型，通过神经肌肉电刺激肌肉训练

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional ...

科研

医学

Cardiac Stress Test Induced by Dobutamine and Monitored by Cardiac Catheterization in Mice

多巴酚丁胺诱导的心脏压力测试和监控通过在小鼠心脏导管插入术

Dobutamine is a &beta;-adrenergic agonist with an affinity higher for receptor expressed in the heart (&beta;1) than for receptors expressed in the ...

Measurement of the Pressure-volume Curve in Mouse Lungs

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential ...

Cardiac Catheterization in Mice to Measure the Pressure Volume Relationship: Investigating the Bowditch Effect

心导​​管在小鼠测量压力体积的关系：调查的影响鲍迪奇

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these ...

Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice

小鼠急性心肌梗死的实时压力容积分析

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any ...

Hemocompatibility Testing of Blood-Contacting Implants in a Flow Loop Model Mimicking Human Blood Flow

流循环模型中血液接触植入物的血液相容性测试模拟人类血流

The growing use of medical devices (e.g., vascular grafts, stents, and cardiac catheters) for temporary or permanent purposes that remain in the ...

Biventricular Assessment of Cardiac Function and Pressure-Volume Loops by Closed-Chest Catheterization in Mice

小鼠闭胸导管插入术对心功能和压力-容积环的双心室评估

Assessment of cardiac function is essential to conduct cardiovascular and pulmonary-vascular preclinical research. Pressure-volume loops (PV loops) ...

A Large Animal Model for Acute Kidney Injury by Temporary Bilateral Renal Artery Occlusion

临时双性肾动脉闭塞的急性肾损伤大型动物模型

Acute kidney injury (AKI) is associated with higher risk for morbidity and mortality post-operatively. Ischemia-reperfusion injury (IRI) is the most ...

In vitro Assessment of Cardiac Reprogramming by Measuring Cardiac Specific Calcium Flux with a GCaMP3 Reporter

使用GCaMP3报告器测量心脏特异性钙通量，对心脏重编程进行体外评估

Cardiac reprogramming has become a potentially promising therapy to repair a damaged heart. By introducing multiple transcription factors, including ...

Closed Chest Biventricular Pressure-Volume Loop Recordings with Admittance Catheters in a Porcine Model

猪模型中闭合胸部双心室压力-体积循环记录与导管

Pressure-volume (PV) loop recording enables the state-of-the-art investigation of load-independent variables of ventricular performance. Uni-ventricular ...