Name: a rat lung transplantation model of warm ischemia/reperfusion injury: optimizations to improve outcomes
Uploaded: 2021-10-28T14:00:00
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雄性Sprague-Dawley大鼠（体重180-270克）被商业购买（例如，Envigo），并在俄亥俄州立大学动物设施的无病原体条件下饲养。所有程序均根据NIH和国家研究委员会的实验动物人道护理和使用指南进行人道执行，并经俄亥俄州立大学机构动物护理和使用委员会（IACUC协议#2012A00000135-R2）批准。
1. 初始设置
<ol><li>设置手术设备。 注意：这是一项非生存性手术。如果要进行生存手术?...

<ol>
	<li>Mizuta, T., Kawaguchi, A., Nakahara, K., Kawashima, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+rat+lung+transplantation+using+a+cuff+technique.">Simplified rat lung transplantation using a cuff technique.</a> Journal of Thoracic and Cardiovascular Surgery. 97 (4), 578-581 (1989).</li><li>Zhai, W., 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=Simplified+rat+lung+transplantation+by+using+a+modified+cuff+technique.">Simplified rat lung transplantation by using a modified cuff technique.</a> Journal of Investigative Surgery. 21 (1), 33-37 (2008).</li><li>Goto, 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=Simplified+rat+lung+transplantation+using+a+new+cuff+technique.">Simplified rat lung transplantation using a new cuff technique.</a> Annals of Thoracic Surgery. 93 (6), 2078-2080 (2012).</li><li>Guo, H., 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=Improvements+of+surgical+techniques+in+a+rat+model+of+an+orthotopic+single+lung+transplant.">Improvements of surgical techniques in a rat model of an orthotopic single lung transplant.</a> European Journal of Medical Research. 18, 1 (2013).</li><li>Tian, D., Shiiya, H., Sato, M., Nakajima, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+lung+transplantation+model:+modifications+of+the+cuff+technique.">Rat lung transplantation model: modifications of the cuff technique.</a> Annals of Translational Medicine. 8 (6), 407 (2020).</li><li>Rajab, T. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anastomotic+techniques+for+rat+lung+transplantation.">Anastomotic techniques for rat lung transplantation.</a> World Journal of Transplantation. 8 (2), 38-43 (2018).</li><li>Reis, A., Giaid, A., Serrick, C., Shennib, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+outcome+of+rat+lung+transplantation+with+modification+of+the+nonsuture+external+cuff+technique.">Improved outcome of rat lung transplantation with modification of the nonsuture external cuff technique.</a> Journal of Heart and Lung Transplantation. 14 (2), 274-279 (1995).</li><li>Sugimoto, R., 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=Experimental+orthotopic+lung+transplantation+model+in+rats+with+cold+storage.">Experimental orthotopic lung transplantation model in rats with cold storage.</a> Surgery Today. 39 (7), 641-645 (2009).</li><li>Santana Rodriguez, N., 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=Technical+modifications+of+the+orthotopic+lung+transplantation+model+in+rats+with+brain-dead+donors.">Technical modifications of the orthotopic lung transplantation model in rats with brain-dead donors.</a> Archivos de Bronconeumolog&#237;a. 47 (10), 488-494 (2011).</li><li>Gielis, J. F., 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=A+murine+model+of+lung+ischemia+and+reperfusion+injury:+tricks+of+the+trade.">A murine model of lung ischemia and reperfusion injury: tricks of the trade.</a> Journal of Surgical Research. 194 (2), 659-666 (2015).</li><li>Rajab, T. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Techniques+for+lung+transplantation+in+the+rat.">Techniques for lung transplantation in the rat.</a> Experimental Lung Research. 45 (9-10), 267-274 (2019).</li><li>Iskender, I., 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=Effects+of+Warm+Versus+Cold+Ischemic+Donor+Lung+Preservation+on+the+Underlying+Mechanisms+of+Injuries+During+Ischemia+and+Reperfusion.">Effects of Warm Versus Cold Ischemic Donor Lung Preservation on the Underlying Mechanisms of Injuries During Ischemia and Reperfusion.</a> Transplantation. 102 (5), 760-768 (2018).</li><li>Lin, X., 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=Five-year+update+on+the+mouse+model+of+orthotopic+lung+transplantation:+Scientific+uses,+tricks+of+the+trade,+and+tips+for+success.">Five-year update on the mouse model of orthotopic lung transplantation: Scientific uses, tricks of the trade, and tips for success.</a> Journal of Thoracic Disease. 4 (3), 247-258 (2012).</li><li>Song, J. A., 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=Standardization+of+bronchoalveolar+lavage+method+based+on+suction+frequency+number+and+lavage+fraction+number+using+rats.">Standardization of bronchoalveolar lavage method based on suction frequency number and lavage fraction number using rats.</a> Toxicological Research. 26 (3), 203-208 (2010).</li><li>Chang, J. E., Kim, H. J., Yi, E., Jheon, S., Kim, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+ischemia-reperfusion+injury+in+a+rat+lung+transplantation+model+by+low-concentration+GV1001.">Reduction of ischemia-reperfusion injury in a rat lung transplantation model by low-concentration GV1001.</a> European Journal of Cardio-Thoracic Surgery. 50 (5), 972-979 (2016).</li><li>De Backer, J. W., 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=Study+of+the+variability+in+upper+and+lower+airway+morphology+in+Sprague-Dawley+rats+using+modern+micro-CT+scan-based+segmentation+techniques.">Study of the variability in upper and lower airway morphology in Sprague-Dawley rats using modern micro-CT scan-based segmentation techniques.</a> Anatomical Record. 292 (5), 720-727 (2009).</li><li>Nelson, K., 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=Method+of+isolated+ex+vivo+lung+perfusion+in+a+rat+model:+lessons+learned+from+developing+a+rat+EVLP+program.">Method of isolated ex vivo lung perfusion in a rat model: lessons learned from developing a rat EVLP program.</a> Journal of Visualized Experiments. (96), e52309 (2015).</li><li>Suzuki, H., Fan, L., Wilkes, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+obliterative+bronchiolitis+in+a+murine+model+of+orthotopic+lung+transplantation.">Development of obliterative bronchiolitis in a murine model of orthotopic lung transplantation.</a> Journal of Visualized Experiments. (65), e3947 (2012).</li><li>Jia, Y., 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=Treatment+of+acute+lung+injury+by+targeting+MG53-mediated+cell+membrane+repair.">Treatment of acute lung injury by targeting MG53-mediated cell membrane repair.</a> Nature Communications. 5, 4387 (2014).</li><li>Kim, J. 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=Pegylated-Catalase+Is+Protective+in+Lung+Ischemic+Injury+and+Oxidative+Stress.">Pegylated-Catalase Is Protective in Lung Ischemic Injury and Oxidative Stress.</a> Annals of Thoracic Surgery. , (2020).</li><li>Beal, E. W., 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=D-Ala(2),+D-Leu(5)]+Enkephalin+Improves+Liver+Preservation+During+Normothermic+Ex+Vivo+Perfusion.">D-Ala(2), D-Leu(5)] Enkephalin Improves Liver Preservation During Normothermic Ex Vivo Perfusion.</a> Journal of Surgical Research. 241 (2), 323-335 (2019).</li><li>Akateh, 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=Intrahepatic+Delivery+of+Pegylated+Catalase+Is+Protective+in+a+Rat+Ischemia/Reperfusion+Injury+Model.">Intrahepatic Delivery of Pegylated Catalase Is Protective in a Rat Ischemia/Reperfusion Injury Model.</a> Journal of Surgical Research. 238, 152-163 (2019).</li></ol>

在本报告中，我们在大鼠肺移植方案中的几个关键步骤进行了干预，以优化程序。虽然已经报道了用于大鼠肺移植的各种袖带技术1，2，3，4，5，6，7，8，9，15，但该过程仍然是主观的...

三十多年来，研究人员一直在修改和改进大鼠肺移植模型，以使生成的数据更加一致，更能反映实际临床状况。在我们实验室执行该模型的过程中，我们确定了四个改进领域：吻合的袖带技术，识别受体的第4肋间隙， 接受者手术过程中的肺充气和伤口闭合，以及收集样本进行分析。
吻合口的袖带技术修改可以通过缩短供体肺1，2，3，4，5，6的处理时间并使显微外科医生的吻合手术更快，技术上更容易来改善整个移植过程。虽然使用适当尺寸的袖带为移植肺提供必要的血液和气流至关重要，但关于如何为肺静脉 （PV）、肺动脉 （PA） 或支气管 （Br） 选择袖带尺寸的指导有限5，7，8，9。由于PV，PA和Br的直径与供体和受体大鼠的体重有关，我们建议袖带尺寸基于体重。该报告根据大鼠的体重（180克至270克以上）提供了袖带的尺寸指南，用于优化移植肺的血液和空气供应（表1）。
虽然较新的显微外科医生可以在供体手术期间成功且轻松地获得供体肺，但在接受者的手术过程中移植肺更为复杂，并且取决于显微外科医生的经验。尝试找到第 4 个肋间隙以进入接受者的左肺是更困难的步骤之一，具有一定的主观性并可能增加手术时间。因此，我们介绍了一种简单客观的方法，通过胸部测量和心脏的心悸来辅助识别第4肋间隙位置，以找到正确的胸壁区域来解剖4，5，6，10，11，12。
我们还建议改善供体肺充气，这是器官损伤的潜在来源。供体肺放气，直到再灌注开始。在缝合第 4 个肋间隙时，供体肺通常通过将 PEEP 从 2 cmH 2 O 增加到 6 cmH2O 来充气。为了尽量减少过度充气造成的肺损伤，我们提出了一种技术，即在低于第5肋的第4肋周围放置三个6-0尼龙缝合线，并带有简单的双结。当需要缝合伤口时，双手用止血器握住三条缝合线的末端，通过向两侧拉动伤口一次闭合，PEEP立即降低至2cmH2O。通过这种方式，允许肺在尽可能短的时间内扩张10。
在实验结束时，研究人员通常希望从每次移植中收集多种类型的样本以进行多种类型的分析。例如，快速冷冻组织、福尔马林固定组织、用于确定肺水肿的干湿重量比的组织以及支气管软体灌洗液 （BAL） 都可用于评估移植进展。收集BAL液体的传统方法允许从受体的原生叶和供体的移植叶13，14，15混合混合样本。为了克服这个问题，我们提出了一种夹住肺门区域的方法，该方法可以更精确地了解移植和天然肺的状况。此外，用于从肺部两侧收集BAL液的PBS体积很重要，因为BAL液含有许多可溶性因子，例如通过浓度测量的细胞因子和趋化因子。将输注的液体量与估计的肺活量恢复正常化有助于比较。右侧有四个裂片，左侧有一个裂片，大鼠的五个裂片中的每一个都有不同的体积和表面积16。根据Backer等人之前对肺叶体积测量的研究，在整个肺的总体积中，右叶的体积为63%（4400mm 3），左叶的体积为37%（2500mm3）。因此，我们建议用于收集BAL液体的PBS体积应计算为右肺潮气量（7.2mL/kg）乘以63%和左肺37%的两倍。通过使用这种方法，可以更好地控制体重和时间10，16 等变量。
总之，在本报告中，我们将展示对大鼠肺移植标准实验模型的一些修改，这些修改可以使程序更有效，并提高从每个实验中生成更准确和更丰富的数据的能力。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>12 Gauge angio-catheter</td>
			<td>BD</td>
			<td>382277</td>
			<td></td>
		</tr>
		<tr>
			<td>14 Gauge angio-catheter</td>
			<td>B. Braun</td>
			<td>4251717-02</td>
			<td></td>
		</tr>
		<tr>
			<td>16 Gauge angio-catheter</td>
			<td>B. Braun</td>
			<td>4252586-02</td>
			<td></td>
		</tr>
		<tr>
			<td>18 Gauge angio-catheter</td>
			<td>B. Braun</td>
			<td>4251679-02</td>
			<td></td>
		</tr>
		<tr>
			<td>20 Gauge angio-catheter</td>
			<td>B. Braun</td>
			<td>4252527-02</td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 silk suture</td>
			<td>Surgical Specialties Corp.</td>
			<td>SP116</td>
			<td></td>
		</tr>
		<tr>
			<td>6-0 nylon suture</td>
			<td>AD Surgical</td>
			<td>S-N618R13</td>
			<td></td>
		</tr>
		<tr>
			<td>7-0 nylon suture</td>
			<td>AD Surgical</td>
			<td>S-N718SP13</td>
			<td></td>
		</tr>
		<tr>
			<td>8-0 nylon suture</td>
			<td>AD Surgical</td>
			<td>XXS-N807T6</td>
			<td></td>
		</tr>
		<tr>
			<td>Betadine Spray</td>
			<td>Avrio Health L.P</td>
			<td>UPC 367618160039</td>
			<td></td>
		</tr>
		<tr>
			<td>Clippers</td>
			<td>VWR</td>
			<td>MSPP-023326</td>
			<td></td>
		</tr>
		<tr>
			<td>Castroviejo micro dissecting spring scissors</td>
			<td>Roboz Surgical Instrument Co</td>
			<td>RS-5668</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #5 - Fine Forceps</td>
			<td>Fine Science Tools</td>
			<td>11254-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrocautery</td>
			<td>Macan</td>
			<td>MV-7A</td>
			<td></td>
		</tr>
		<tr>
			<td>Endotracheal intubation kit</td>
			<td>Kent Scientific</td>
			<td>ETI-MSE</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>Fine Science Tools</td>
			<td>11027-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Halsted-mosquito hemostat</td>
			<td>Roboz Surgical Instrument Co</td>
			<td>RS-7112</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin</td>
			<td>Fresnius Medical Care</td>
			<td>C504701</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin syringe</td>
			<td>Life Technologies</td>
			<td>B328446</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Piramal Critical Care</td>
			<td>NDC 66794-017-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropyl Alcohol Swabs</td>
			<td>BD</td>
			<td>326895</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Hikma Pharmaceuticals PLC</td>
			<td>NDC 0413-9505-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Dieffenbach Bulldog Clamp</td>
			<td>World Precision Instruments</td>
			<td>WPI14117</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder/Forceps, Curved</td>
			<td>Micrins</td>
			<td>MI1542</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder/Forceps, Straight</td>
			<td>Micrins</td>
			<td>MI1540</td>
			<td></td>
		</tr>
		<tr>
			<td>Perfadex Plus (Organ Preservation Solution)</td>
			<td>XVIVO Perfusion AB</td>
			<td>REF# 19950</td>
			<td></td>
		</tr>
		<tr>
			<td>PhysioSuite</td>
			<td>Kent Scientific</td>
			<td>PS-MSTAT-RT</td>
			<td>Used to check SpO2 and heartbeat</td>
		</tr>
		<tr>
			<td>Retractor</td>
			<td>Roboz Surgical Instrument Co</td>
			<td>RS-6560</td>
			<td></td>
		</tr>
		<tr>
			<td>Saline</td>
			<td>PP Pharmaceuticals LLC</td>
			<td>NDC 63323-186-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Fine Science Tools</td>
			<td>14090-11</td>
			<td></td>
		</tr>
		<tr>
			<td>SomnoSuite Small Animal Anesthesia System</td>
			<td>Kent Scientific</td>
			<td>SS-MVG-Module</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile&#160; Cotton Gauze Pad</td>
			<td>Fisherbrand</td>
			<td>22-415-469</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical Microscope</td>
			<td>Leica</td>
			<td>M500-N w/ OHS</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe 5mL</td>
			<td>BD</td>
			<td>309646</td>
			<td></td>
		</tr>
		<tr>
			<td>Vannas-Tubingen Spring Scissors</td>
			<td>Fine Science Tools</td>
			<td>15008-08</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Korn Pharmaceuticals Corp</td>
			<td>NDC 59399-110-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Yasargil Clamp</td>
			<td>Aesculap, Inc</td>
			<td>FT351T</td>
			<td>Used to clamp bronchus</td>
		</tr>
		<tr>
			<td>Yasargil Clamp</td>
			<td>Aesculap, Inc</td>
			<td>FT261T</td>
			<td>Used to clamp hilum</td>
		</tr>
		<tr>
			<td>Yasargil Clamp Applicator</td>
			<td>Aesculap, Inc</td>
			<td>FT484T</td>
			<td></td>
		</tr>
	</tbody>
</table>

a rat lung transplantation model of warm ischemia/reperfusion injury: optimizations to improve outcomes

根据我们在大鼠肺移植方面的经验，我们发现有几个需要改进的领域。现有文献中关于为肺静脉 （PV）、肺动脉 （PA） 或支气管 （Br） 选择合适的袖带尺寸的方法的信息各不相同，因此在大鼠肺移植期间确定适当的袖带尺寸是一项反复试验。通过标准化袖带技术以使用适合血管或支气管大小的最小有效袖带，可以使移植过程更安全、更快、更成功。由于PV，PA和Br的直径与大鼠的体重有关，因此我们提出了一种使用基于重量的指南选择合适的尺寸的策略。由于肺容量也与体重有关，我们建议在为温暖缺血期间供体肺充气选择合适的空气量以及在支气管肺泡灌洗（BAL）液体收集期间滴注适当体积的PBS时，也应考虑这种关系。我们还描述了第 4肋 间隙解剖、伤口闭合以及从天然和移植叶收集样本的方法。

为了测量肺水肿，计算湿与干重比。按照协议中的描述收集供体的原生叶，移植的叶和受体的天然叶，并立即称量湿重，在60°C干燥48小时，然后再次称量干重。湿重比增加提示肺水肿。我们的结果表明，与供体或受体的原生叶相比，移植叶的干湿重量比确实显着增加（p = 0.0050，n = 6 /组; 图 11）。
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A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes Video (Video) | JoVE

在这里，我们提出了对大鼠肺移植模型的优化，以改善结果。我们提供基于体重的袖带尺寸指南、确定第 4 肋 间隙的测量策略以及伤口闭合和 BAL（支气管肺泡灌洗）液体和组织收集的方法。

温缺血/再灌注损伤的大鼠肺移植模型：改善结局的优化

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...

该方案确定了四个关键领域，以帮助改进方法并获得可靠的肺移植。首先是可重复的袖带技术。其次是准确且可重复地识别第四肋间隙，彻底闭合伤口，然后如何恢复和获取样品进行分析。这些技术有助于开发一种可靠的肺移植方法，以研究分子机制并开发治疗干预措施。首先设置手术设备，打开心率/血氧饱和度监测设备并将电路板预热至 42 摄氏度。另外，打开通气和麻醉机以预热异氟醚蒸发器。用10毫升液体异氟醚填充麻醉注射器，并将注射器安装到通气和麻醉机上。接下来，打开手术显微镜，根据喜好调整高度和焦点并启动电灼装置。同样，准备和布置手术工具。通过参考表1，根据大鼠的体重确定用于制作袖带的血管导管的正确规格尺寸。准备好肺动脉、支气管和肺静脉的袖带后，将适当大小的血管导管放在手术显微镜下的无菌表面上。然后，使用11号肋背手术刀片将血管导管切割成90度角，形成一个两毫米长的袖带体，袖带主体顶部的宽度和高度为1毫米。准备好手术包裹后，用剪刀在颈部中部做一个三到四厘米长的中线皮肤切口，用镊子小心地解剖皮下组织和肌肉。对于气管插管，在气管周围穿一条 4-0 丝缝线。然后，将 16 号血管导管插入气管，并用双结将气管周围的缝合线紧紧地绑住，然后打一个结以将血管导管固定到位。将血管导管连接到呼吸机，以维持大鼠用1至2%异氟醚的手术麻醉平面。使用剪刀进行腹腔胸骨切开术，作为中线和横向切口的组合。然后，通过下腔静脉或IVC用胰岛素注射器注射每公斤肝素1， 000单位，并允许10分钟进行体循环。当肺部通气时，用微型解剖弹簧剪刀切割左右耳廓，然后通过肺动脉引入 18 号血管导管，在悬挂在 28 厘米水中的注射器中用 20 毫升保存溶液重力冲洗肺部，连接到管道和血管导管。断开呼吸机与气管插管的连接，以连接到根据体重填充适当空气量的 5 毫升注射器。温暖缺血一小时后，用微型解剖弹簧剪刀和镊子切除心肺阻滞，并放在用冰冷的PBS蘸湿的无菌纱布上，放在冰上的无菌培养皿上。用Vannas-Tubingen弹簧剪刀小心修剪左肺门区域，并获取左肺静脉，肺动脉和支气管。使用蚊子止血器抓住袖带标签。然后，将肺静脉、肺动脉或支气管的远端通过适当的袖带体，并清除袖带周围的额外组织，然后用 8-0 固定尼龙缝合线。完成后，使用Vannas-Tubingen弹簧剪刀修剪袖带身体周围的多余组织和袖带。为了准备受体大鼠，在大鼠的胸部从胸骨上切口到剑突画一条线。在这条线的中间，沿着胸部左侧画一条线，测量从胸骨上切口到剑突的一半测量值。使用连接到气管插管套件的 LED 灯的光缆，使用 16 号血管导管观察并插管接受者。一旦肺被拔出，在左肺周围放置无菌纱布，同时用迪芬巴赫斗牛犬夹住它，并在左肺门区域尽可能近端应用Yasargil夹。当大鼠准备好时，通过在受体的肺静脉周围放置7-0尼龙缝合线来进行肺静脉吻合。使用Vannas-Tubingen弹簧剪刀切开接受者的肺静脉，尽可能远端横向切割上段和下段静脉，并使用胰岛素注射器用每毫升肝素化盐水0.2毫升一个国际单位冲洗血液。将用冰冷、湿、无菌纱布包裹的供体肺放入胸腔，并将供体的袖带囊肺静脉插入受者的肺静脉，然后用预先定位的 7-0 尼龙缝合线固定吻合口。对于支气管或支气管吻合术，在通过横向切开上段和下节段气道尽可能远方切开受者的支气管之前，在受体的支气管周围放置 7-0 尼龙缝合线。将供体的带袖口支气管插入接受者的支气管后，用预先定位的 7-0 尼龙缝合线固定吻合口。接下来，如前所述进行肺动脉吻合术。为了允许移植的供体肺的再灌注和通气，取下肺门上的Yasargil钳，然后用6-0尼龙缝合线关闭开胸切口。根据需要从天然或移植的肺中收集BAL液。计算肺叶干湿重比以测量肺水肿。与供体或受体的自体叶相比，移植叶的湿重与干重比显着增加，表明没有肺水肿。在我们看来，该程序的两个关键步骤是，一，准确地使用解剖标志来识别第四肋间隙，然后二，选择并适当使用袖带以具有可重复的袖带技术。既可重复又一致的大鼠肺移植模型是缺血再灌注和一般移植的分子研究的基础。

Research

JoVE Journal

Medicine

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

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常温心脏骤停和心肺复苏术：缺血再灌注损伤的小鼠模型

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
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科研

医学

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In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

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在原位横向腹直肌肌皮瓣：肌缺血再灌注损伤模型大鼠

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急性肾损伤和伤后纤维化小鼠缺血再灌注模型

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

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心肌缺血再灌注损伤的通过左前降支结扎小鼠模型

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

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的隔离方法<em&gt;离体</em&gt;肺灌注的大鼠模型：教训制定鼠EVLP课程中学

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诱导和缺血再灌注损伤评估中的Langendorff灌注鼠心

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

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肠缺血再灌注损伤的小鼠模型

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

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视网膜缺血再灌注损伤的小鼠模型通过眼压升高

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

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大鼠心肌缺血再灌注后冠状动脉流动储备评价

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