This study was supported by the Netherlands Cardiovascular Research Initiative, the Dutch Heart Foundation, the Dutch Federation of University Medical Centers, the Netherlands Organization for Health Research and Development, and the Royal Netherlands Academy of Sciences (CVON nr. 2012-08, PHAEDRA, The Sebald fund, Stichting Hartekind).

涉及受试动物的程序已通过荷兰中央委员会的动物实验和动物保健委员会在格罗宁根大学医学中心（NL）。使用了这两个的Wistar和刘易斯大鼠180和300克之间的权。 1.住房和驯化<ol><li>在中央动物设施，房屋大鼠的每笼5组到任后。在7天的适应期后，习惯于老鼠对人类的处理，但不进行任何实验程序。 </li></ol> 2.准备和无菌Monocrotalin注射<ol><li>为1毫升60毫克/毫升monocrotalin（MCT）溶液，称重60毫克monocrotalin的在2毫升管中。添加0.9％的NaCl的700微升。加入1 M盐酸200微升。温热下热运转的自来水和涡流它在管中的溶液。使用6N的氢氧化钠以使pH向7.0。使用无菌技术制备的MCT的用于注入啮齿动物。 </LI&gt; <li>注入无菌60毫克/毫升的MCT每kg皮下颈部溶液（0.3毫升的60毫克/毫升的MCT为一个300-G大鼠）1毫升。注意:我们不喜欢使用更小的体积由于机会较大的注射剂量不恰当的。 </li></ol> 3.主动脉腔静脉分流手术<ol><li>麻醉。 <ol><li>填用5％异氟烷/ 100％O 2的感应腔（流速:1升/分钟），然后将鼠在腔室。通过执行后脚趾捏检查麻醉深度足够。称量老鼠。 </li><li>刮脸和清洁腹部过了约8厘米长，宽3厘米的区域。放置老鼠在其上通过一个无菌垫覆盖的热垫（37℃）背部。 </li><li>放置吻在通气面罩/罩2 - 3％异氟烷/ 100％O 2（流速:1升/分钟）。通过执行后趾压紧检查麻醉深度。应用眼膏，以防止干燥，而在麻醉下。 </li></ol></li><li>分流苏尔格里。 <ol><li>擦洗氯化物hexidine消毒皮肤。注入0.01毫克/千克丁丙诺啡皮下术后镇痛。 </li><li>使用无菌器械进行手术。作出切口在腹部＃10解剖刀刀片上的中线，在开始向下延伸到正好在生殖器上述膈下1厘米。 </li><li>抬起用棉签肠道，盖在无菌，湿纱布（0.9％NaCl）中的肠，并把它们放置在动物的左侧。 </li><li>用棉签到附加腹主动脉和下腔静脉周围组织的膜分离。 注意:不要解剖主动脉和下腔静脉之间的隔膜。 </li><li>使用分裂镊子，去除血管周围主动脉脂肪只是分叉以上，仅在主动脉的右侧，并且只在其中针将被插入的位点。 </li><li>用棉签主动脉和下腔静脉由2毫米分离苏佩rior到针会为了要Biemer钳创造空间被插入的位点。 </li><li>在这个区域中，首先放置一个松散结扎（5-0缝合）围绕主动脉。通过放置一个科赫尔钳在其上创建的结扎张力，然后将科赫尔优于切口（ 图1A）。放置Biemer钳只是优于结扎（FIGURē1A）。 </li><li>用棉签，如向远端尽可能压缩腔静脉阻塞的流动（ 图1A）。弯针（18克在这个协议）成45度角，与朝外面（ 图1A）孔口指点。 </li><li>以90度的角度，插入针在主动脉，只是分叉上方，与针指向的孔向左侧（ 图1A）。操纵针向左的前端，将其插入腔静脉。 注:针尖现在应该VISI竹叶提取在腔静脉（ 图1B）。 </li><li>使用第二棉签推剩余的血液在主动脉出插入位点的，以防止血栓形成。干周围用无菌纱布分流的区域，以便使胶充分粘。 </li><li>拉动整个针从主动脉并立即应用组织胶一滴到在主动脉穿刺部位。确保不要棉签粘上来组织。松开主动脉。 </li><li>拉动和释放的主动脉近端分流结扎手动验证分流。松动应上色腔静脉远端鲜红色分流和分流站点上创建的动荡。 注:紧缩将会把血液中的腔静脉回暗红色。 </li><li>放置肠放回动物。关闭肌肉层和皮肤可吸收缝线4-0。通风用100％O 2的动物从麻醉中恢复。 注意:不要让一个ANI发作无人值守，直到它已经恢复了足够的意识，以保持胸骨斜卧。 </li></ol></li><li>假手术。 <ol><li>执行上述所有的方法，只是在插入针的进入主动脉。 </li></ol></li><li>手术后的护理。 <ol><li>放置老鼠在单个笼和入在37℃的培养箱，直到第二天早上。 </li><li>周围手术后6小时，注入0.01毫克/千克丁丙诺啡皮下术后镇痛。如果老鼠显示不适的迹象，第二天早上重复。 注意:手术后第3天，老鼠喜欢吃少喝（当饲料及饮水与药物混合，这一点尤为重要）。大多数老鼠显示手术后第3天的正常行为。如果没有，密切监测。 1周后超过15％的重量损失被认为是不正常的，并且这样的大鼠应在循环血液量而麻醉下的提取安乐死。 </li></ol></li></ol>注意:在这个协议中，该动物是由循环血液量而麻醉下的提取安乐死。 <ol><li>手术（MF8）对早期细胞和功能性反应肺血流增加（ 例如，基因上调或早期转录因子）后牺牲1天。 </li><li>手术（MF14）为早期PAH血管表型（膜肥厚内膜无损伤）后牺牲1周。 </li><li>手术（MF21）对于晚期PAH血管表型（标记膜肥厚和内膜增生）与RVP和肺动脉压轻度升高后牺牲2周。 </li><li>手术（MF28）为终末期PAH血管型（标内膜闭塞）和强大的海拔RVP和肺动脉压后牺牲3周。右心衰竭的临床症状在这个阶段很常见。 </li><li>第28天（MF-RVF后牺牲）对PAH相关的右心室衰竭（RVF），临床上定义为呼吸困难，严重嗜睡，体重下降（&lt;1周10％）。终止时，老鼠的这些迹象之一是存在。通常情况下，老鼠患天28和35之间，这些症状，如果无人看管，在该时间间隔期间死亡油然而生。 </li></ol> 5.右心导管<ol><li>麻醉。 <ol><li>填用5％异氟烷/ 100％O 2的感应腔（流速:1升/分钟），然后将鼠在框中。通过执行后脚趾捏检查麻醉深度足够。称量老鼠。 </li><li>剃和在大鼠的右腹侧清洁颈部和，对于心动协议，胸部和上腹部。 </li><li>放置在其上的热垫（37℃）背面的大鼠和放置鼻子在通气面罩/罩2 - 3％异氟烷/ 100％O 2（流速:1升/分钟）。吻应该面对朝研究员。 </li><li&gt;检查麻醉深度。小心大鼠重症PH。如果心脏速率降低，减少麻醉深度。优选的，执行20分钟之内的所有的测量。应用眼膏，以防止干燥，而在麻醉下。 </li></ol></li><li>超声心动图的协议。 <ol><li>根据由布里泰恩等人描述的协议来执行超声心动图。在朱庇特13。 </li></ol></li><li>导尿协议。 注意:此协议使用一个预成形端部的刚性套管弯曲20度，以指导15厘米硅导管 ，从尖端球2毫米。与其孔口微曲到内部的20-G 针用于插入套管插入右颈静脉（见材料的列表）。在PAH进展和控制的任何阶段只可在该协议中使用。 <ol><li>用消毒氯hexidine颈部。做一个1.5厘米的切口，在右腹侧＃10手术刀刀片颈部，从右侧锁骨到下颌骨。 </li><li>用剪刀传播组织。使用镊子，轻轻直到出现颈静脉拉开组织。周围解剖颈膜使用静脉分裂镊子。 </li><li>通过放置一个松散的结扎（缝合5-0）船舶周围戴上颈静脉的张力。加剧紧张局势，并用胶带将结扎到通气面罩（ 图 2A）。 </li><li>下游的插入位点，放置一个松散结扎血管周围收紧后套管是在原地 ，以防止泄漏和压力损失。 </li><li>使用镊子的把手，稍微弯曲的孔的内部的20-G针的前端与导管进行插管。 </li><li>引入20-G针插入静脉的尖端，并迅速放入含有容器内的导管插管。拔出针头，然后关闭结扎已在步骤5.3.4准备。 </li><li>进行含有导管插入颈静脉插管。套管的前端是在20度曲线（见步骤5.3.5）。操纵锁骨下套管和前进少许进入右心房（ 图2C）。 </li><li>进入右心室，指向套管向左的前端，朝向心脏（ 图2D）。在床旁监护仪，一个RV压力曲线应该出现，匹配图2D。 </li><li>当RV压力曲线是恒定的，写下收缩和舒张右心室压力1（sRVP1 / dRVP1）。 </li><li>操纵套管的左侧和向上的尖端。套管（ 图2E）内推进导管。 </li><li>推进导管进入主肺动脉（PA）。经过肺动脉瓣时，应感到无阻力。 注:当导管进入主肺动脉，则Diastolic压力将上升。在床边监护仪，一个PA压力曲线应该出现，配套图2E。 </li><li>当PA压力曲线是恒定的，记下收缩压，舒张压，平均PA的压力1（sPAP1，dPAP1，mPAP1）。 </li><li>进一步推进套管内的导管，直到导管的前端的球在肺动脉被楔入。观察床旁监护仪下降压力曲线和匹配图2F楔形压力曲线。 </li><li>当楔压曲线是恒定的，记下收缩压，舒张压，平均楔压。 </li><li>拉回导管慢慢地，随后测量并记下sPAP2，dPAP2，mPAP2，sRVP2和dRVP2值，床旁监护仪上显示。 </li><li>当房车，略有回拉套管和导管测量平均右房压（RAP）。曲线应该与图2A的RAP曲线。 没有TE:在这个协议中，将大鼠的导尿协议之后，由循环血液量的同时麻醉下提取安乐死。 </li></ol></li></ol> 6.形态学评估和形态测量注意:在这个协议中，该动物是由循环血液量而麻醉下的提取安乐死。在PAH进展和控制的任何阶段只可在该协议中使用。 <ol><li>处死后，通过切割约5mm支气管分叉以上和肺连接到心脏血管气管取出肺。把肺部冷盐水。解剖左肺。切在分叉的左主支气管。 </li><li>填写一个50毫升的注射器，用4％多聚甲醛，附加一个管套管注射器，并悬挂约工作台上方一米的注射器。适合在左主支气管插管被动填充用多聚甲醛的肺。手柄谨慎多聚甲醛。 </li><li>孵育在多聚甲醛左肺48小时。 </li><li>通过在70％乙醇（1小时），80％乙醇（1小时），90％乙醇（1小时），100％乙醇（3小时），混合二甲苯（2小时），和石蜡连续孵育它脱水左肺（ 2小时）。 </li><li>嵌入左肺石蜡，面向磁带肺肺门。 </li><li>染色用维尔赫夫或弹性van Gieson染色染色的石蜡包埋，4微米肺切片，按照制造商的说明29。确保弹性薄片是公分化（ 如图3）。扫描染色切片放大40倍。 </li><li>划分成肺4个象限。在每一个象限中，找到10容器的外径&lt;50微米（帧内腺泡）和10容器的外径&gt; 50微米（预腺泡）。拍照（每肺2×40图片）。放大随机高达20倍的放大倍率和照片每艘船在视这一领域，以尽量减少选择偏差。 </li><li>排除具有&gt; 2最长/最短径比，一个不完整的圆形，或血管壁的四分之一以上的崩溃血管。 注:排除的容器的一个例子显示于图3b中制作的相同的放大倍率（40X）每一画面，并包括一个比例尺。 </li><li>打开ImageJ的与第一张照片。绘制比例尺的直线上的图象通过" 分析 "，设置刻度 和"设置规模 。" 素以" 距离"，使用图片的比例尺上的值。用微米（μm），为长度单位。将比例设置为全局。 </li><li>使用" 写意选择 ，" 画一条线上的管腔面积（ 图3）的内边界， 并使用"测量 " （CRTL M）来衡量这个区域。然后，周围画外ELAST线集成电路层（ 图3）来测量总血管面积。 </li><li>计算管腔和外径（ <img alt="方程" src="/files/ftp_upload/55065/55065eq1.jpg" /> ）使用<img alt="方程" src="/files/ftp_upload/55065/55065eq2.jpg" /> 。 </li><li>使用计算壁厚<img alt="方程" src="/files/ftp_upload/55065/55065eq3.jpg" /> 。 </li><li>通过计算墙/管腔比<img alt="方程" src="/files/ftp_upload/55065/55065eq4.jpg" /> 。 </li><li>使用计算遮挡比分<img alt="方程" src="/files/ftp_upload/55065/55065eq5.jpg" /> 。 </li><li>得分上肌在容器（无，部分，或总肌）（ 图3B）。 注:船只具有双重弹性薄片的一半以上的圆周被定义为完全肌型。具有双弹性薄片不到一半的圆周血管被定义为部分肌型。 </li><li>得分上neoi的存在下容器ntima（是或否）（ 图3C）。 注:船舶没有明确定义的内弹力层与（通常偏心）管腔闭塞合并被定义为内膜病变。 </li></ol>

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G., Bartelds, B., Molema, G., 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=Egr-1+expression+during+neointimal+development+in+flow-associated+pulmonary+hypertension.">Egr-1 expression during neointimal development in flow-associated pulmonary hypertension.</a> Am J Pathol. 179 (5), 2199-2209 (2011).</li><li>Borgdorff, M. A., Bartelds, B., Dickinson, M. G., Steendijk, P., de Vroomen, M., Berger, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+loading+conditions+reveal+various+patterns+of+right+ventricular+adaptation.">Distinct loading conditions reveal various patterns of right ventricular adaptation.</a> Am J Physiol Heart Circ Physiol. 305 (3), H354-H364 (2013).</li><li>Ruiter, G., de Man, F. S., Schalij, 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=Reversibility+of+the+monocrotaline+pulmonary+hypertension+rat+model.">Reversibility of the monocrotaline pulmonary hypertension rat model.</a> Eur Respir J. 42 (2), 553-556 (2013).</li><li>van Albada, M. E., Bartelds, B., Wijnberg, 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=Gene+expression+profile+in+flow-associated+pulmonary+arterial+hypertension+with+neointimal+lesions.">Gene expression profile in flow-associated pulmonary arterial hypertension with neointimal lesions.</a> Am J Physiol Lung Cell Mol Physiol. 298 (4), L483-L491 (2010).</li><li>Dickinson, M. G., Kowalski, P. S., Bartelds, B., 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+critical+role+for+egr-1+during+vascular+remodelling+in+pulmonary+arterial+hypertension.">A critical role for egr-1 during vascular remodelling in pulmonary arterial hypertension.</a> Cardiovasc Res. 103 (4), 573-584 (2014).</li><li>van der Feen, D. E., Dickinson, M. G., Bartelds, M. G., 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=Egr-1+identifies+neointimal+remodeling+and+relates+to+progression+in+human+pulmonary+arterial+hypertension.">Egr-1 identifies neointimal remodeling and relates to progression in human pulmonary arterial hypertension.</a> Jheart lung transplant. 35 (4), 481-490 (2016).</li><li>Rungatscher, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+overcirculation-induced+pulmonary+arterial+hypertension+in+aorto-caval+shunt.">Chronic overcirculation-induced pulmonary arterial hypertension in aorto-caval shunt.</a> Microvasc Res. 94, 73-79 (2014).</li><li>O'Blenes, S. B., Fischer, S., McIntyre, B., Keshavjee, S., Rabinovitch, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemodynamic+unloading+leads+to+regression+of+pulmonary+vascular+disease+in+rats.">Hemodynamic unloading leads to regression of pulmonary vascular disease in rats.</a> J Thorac Cardiovasc Surg. 121 (2), 279-289 (2001).</li><li>Sakao, S., Taraseviciene-Stewart, L., Lee, J. D., Wood, K., Cool, C. D., Voelkel, N. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Initial+apoptosis+is+followed+by+increased+proliferation+of+apoptosis-resistant+endothelial+cells.">Initial apoptosis is followed by increased proliferation of apoptosis-resistant endothelial cells.</a> FASEB J. 19 (9), 1178-1180 (2005).</li><li>Spiekerkoetter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FK506+activates+BMPR2,+rescues+endothelial+dysfunction,+and+reverses+pulmonary+hypertension.">FK506 activates BMPR2, rescues endothelial dysfunction, and reverses pulmonary hypertension.</a> J Clin Invest. 123 (8), 3600-3613 (2013).</li><li>Nickel, N. P., Spiekerkoetter, E., Gu, 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=Elafin+reverses+pulmonary+hypertension+via+caveolin-1-dependent+bone+morphogenetic+protein+signaling.">Elafin reverses pulmonary hypertension via caveolin-1-dependent bone morphogenetic protein signaling.</a> Am J Respir Crit Care Med. 191 (11), 1273-1286 (2015).</li><li>Meloche, J., Potus, F., Vaillancourt, 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=Bromodomain-containing+protein+4:+The+epigenetic+origin+of+pulmonary+arterial+hypertension.">Bromodomain-containing protein 4: The epigenetic origin of pulmonary arterial hypertension.</a> Circ Res. 117 (6), 525-535 (2015).</li><li>Happ&#233;, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pneumonectomy+combined+with+SU5416+induces+severe+pulmonary+hypertension+in+rats.">Pneumonectomy combined with SU5416 induces severe pulmonary hypertension in rats.</a> Am J Physiol Lung Cell Mol Physiol. 310 (11), L1088-L1097 (2016).</li><li>Ranchoux, B., Antigny, F., Rucker-Martin, 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=Endothelial-to-mesenchymal+transition+in+pulmonary+hypertension.">Endothelial-to-mesenchymal transition in pulmonary hypertension.</a> Circulation. 131 (11), 1006-1018 (2015).</li><li>de Raaf, M. 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The authors have nothing to disclose.

该方法描述了一种主动脉腔静脉分流的大鼠MCT创建流动引起的肺动脉高压和技术来评估原则血液动力学和表征的PAH和该模型的组织病理学终点预处理的外科手术。 该议定书和故障排除中关键步骤手术和手术后。在主动脉 - 静脉分流手术，最关键的步骤是主动脉和腔静脉的剥离。括住主动脉和腔静脉应足够解剖膜创造1）的主动脉区域，其中，所述针将被插入...

该方法的目标是创造在大鼠为严重，流动引起的肺动脉高压可再现模型，并测量它的原理的血液动力学和组织病理学终点。 肺动脉高压（PAH）是一种临床综合征，它包括肺血管阻力逐步增加，导致右心室衰竭而死亡。内肺动脉高血压疾病（PH）的上级疾病谱，多环芳烃是最严重的形式，另一种仍然没有治愈1。在PAH的根本动脉的特点是血管重塑的一种典型形式，闭塞血管腔。正常的非肌型容器和内侧容器层的肥大的肌被视为在PAH早期疾病的现象，也见于其他形式的PH 2，并且被认为是可逆3。作为一个PAHdvances，内膜层开始改造，最终形成特征性病变内膜2。内膜型肺血管重构是独家PAH，目前认为是不可逆的4。 由于PAH是一种罕见的疾病，在推进其病理学的理解和新疗法的发展严重依赖动物模型。大鼠monocrotalin（MCT）模型是一个简单的单击模式，已经，并且现在仍然是经常使用的。 MCT是导致伤到肺小动脉和地区炎症5毒素。 60毫克/千克的MCT导致增加的平均肺动脉压（mPAP），肺血管阻力（PVR），以及3后右心室肥大（RVH） - 4周5。该组织形态学无内膜病变5的特点是隔离膜肥厚。该MCT从而大鼠模型表示的PH适度的形式，而不是PAH，虽然它通常呈现为后者。 在与先天性左向右分流（PAH-CHD）相关的PAH的孩子，肺血流增加被认为是内膜病变的7，8，9发展的重要触发。在大鼠中，肺血流增加可由创作腹主动脉和腔静脉，在1990年10首先描述的技术之间的分流来诱导。替代创造增加肺血流量是单侧肺或锁骨下肺动脉吻合术11。这些模型的概念缺点包括由肺，或由于肺血管的医源性损伤致残留肺和自适应通路激活潜在补涨肺动脉吻合术，既混淆增加肺血流量的影响。 当创建主动脉腔静脉分流和肺血流增加是诱发作为第二砸在MCT处理的大鼠，发生特点内膜病变，和多环芳烃是一种严重的和相关的右心室衰竭（RVF）制定3周上升后流程12。 PAH的此模型中的血液动力学级数可以在体内通过超声心动图和右心脏导管插入术进行评估。血管组织形态学，血管壁的厚度，小动脉闭塞程度和参数右心室衰竭形成PAH的体外表征的支柱。 这种方法描述了主动脉腔静脉分流术（AC-分流）详细的手术方案，对心脏导管检查和血管组织形态学定性和定量评估。

<table border="0" cellpadding="0" cellspacing="0" width="835">
	<tbody>
		<tr height="25">
			<td height="25" style="height:25px;width:307px;">
			Shunt Surgery
			</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td style="width:196px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sterile surgical gloves</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Duratears Eye ointment</td>
			<td style="width:151px;">Alcon</td>
			<td style="width:183px;">10380</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Chloride-Hexidine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Cotton swabs</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Histoacryllic tissue glue</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td>1050052</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Silkam 5-0 sutures black non-resorbable</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td>F1134027</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Safil 4-0 sutures violet resorbable</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">18 G needle&#160;</td>
			<td style="width:151px;">Luer</td>
			<td>NN1838R BD</td>
			<td>tip bent in 45 degrees orifice to the outside</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Gauzes 10x10 cm</td>
			<td>Paul Hartmann</td>
			<td>407825</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Temgesic Buprenorphine</td>
			<td style="width:151px;">RB Pharmaceuticals</td>
			<td style="width:183px;">5429</td>
			<td>subcutaneous injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sodium Chloride 0.9 %</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Ventilation mask Rat</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Scalple blade</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Biemer clamp 18 mm, 5 mm opening&#160;</td>
			<td><a href="https://www.google.nl/url?sa=i&amp;rct=j&amp;q=&amp;esrc=s&amp;source=images&amp;cd=&amp;cad=rja&amp;uact=8&amp;ved=0ahUKEwitqICpkITNAhWiL8AKHe6iCC8QjhwIBQ&amp;url=http%3A%2F%2Fwww.agnthos.se%2Fquote%2Fquote%2Fcaturl%2Fid%2F242%2FBiemer&amp;psig=AFQjCNEnfRXuMQ6Doi4_-2KMYu3qeRovMA&amp;ust=1464777707579746">AgnTho</a></td>
			<td>64-562</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Heat mat</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Kocher Clamp</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Shaving machine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Microscope</td>
			<td style="width:151px;">Leica</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:307px;">
			Right Heart Catheterization
			</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Name</td>
			<td style="width:151px;">Company</td>
			<td style="width:183px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sterile surgical gloves</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Eye ointment</td>
			<td style="width:151px;">Duratears</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Chloride-Hexidine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Cotton swabs</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Gauzes 10x10 cm</td>
			<td>Paul Hartmann</td>
			<td>407825</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Silkam 5-0 sutures black non-resorbable</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td>F1134027</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Needle 20 G</td>
			<td style="width:151px;">Luer</td>
			<td style="width:183px;"></td>
			<td>Tip slightly bent to the inside</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Cannula 20 G</td>
			<td style="width:151px;">Luer</td>
			<td style="width:183px;"></td>
			<td>to introduce catheter, tip pre-formed in 20 degrees</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Silastic Catheter 15 cm long</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td>0.5 mm ball 2 mm from tip</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Pressure transducer</td>
			<td style="width:151px;">Ailtech</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Bedside monitor Cardiocap/5</td>
			<td style="width:151px;">Datex-Ohmeda</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Shaving machine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">10mL Syringe</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sodium Chloride 0.9 %</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td>for flushing</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:307px;">
			Vascular Morphology
			</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Name</td>
			<td style="width:151px;">Company</td>
			<td style="width:183px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">50ml Syringe</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">4 % Formaldehyde</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">18 G cannula with tube</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Verhoef staining kit</td>
			<td style="width:151px;">Sigma-Aldrich</td>
			<td><a href="http://www.sigmaaldrich.com/catalog/product/sigma/ht254?lang=en&amp;region=NL">HT254</a></td>
			<td>http://www.sigmaaldrich.com/catalog/product/sigma/ht254?lang=en&#38;region=US</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Digital slide scanner</td>
			<td style="width:151px;">Hamamatsu</td>
			<td style="width:183px;">C9600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Image-J</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Elastic (Connective Tissue Stain)&#160;</td>
			<td style="width:151px;">Abcam</td>
			<td style="width:183px;">ab150667</td>
			<td>http://www.abcam.com/elastic-connective-tissue-stain-ab150667.html</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;"></td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td>http://www.abcam.com/ps/products/150/ab150667/documents/ab150667-Elastic%20Stain%20Kit%20(website).pdf</td>
		</tr>
	</tbody>
</table>

shunt surgery, right heart catheterization, and vascular morphometry in a rat model for flow-induced pulmonary arterial hypertension

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 (MCT+Flow). The shunt is created by inserting an 18-G needle from the abdominal aorta into the adjacent caval vein. Increased pulmonary flow has been demonstrated as an essential trigger for a severe form of PAH with distinct phases of disease progression, characterized by early medial hypertrophy followed by neointimal lesions and the progressive occlusion of the small pulmonary vessels. To measure the right heart and pulmonary hemodynamics in this model, right heart catheterization is performed by inserting a rigid cannula containing a flexible ball-tip catheter via the right jugular vein into the right ventricle. The catheter is then advanced into the main and the more distal pulmonary arteries. The histopathology of the pulmonary vasculature is assessed qualitatively, by scoring the pre- and intra-acinar vessels on the degree of muscularization and the presence of a neointima, and quantitatively, by measuring the wall thickness, the wall-lumen ratios, and the occlusion score.

代表性的结果示于图4中。所提出的研究结果表明MCT +流Lewis大鼠在以下群体的特征:（N = 3），MF8（N = 5），MF14（N = 5），MF28（N = 5）和MF-RVF控制（ N = 10）。统计分析采用与Bonferroni校正方差分析单向进行的。 60毫克/千克的MCT和肺血流增加导致收缩期右心室压的平均上升（sRVP）（23±6到56±11毫米汞柱），肺动脉收缩?...

Watch this Scientific Journal Video about Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension at JoVE.com

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

This protocol describes a surgical procedure to create a model for flow-induced pulmonary arterial hypertension (PAH) in rats and the procedures to analyze the principle hemodynamic and histological end-points in this model.

分流手术，右心导管，血管形态计量学在大鼠模型中的流量引起的肺动脉高压

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 ...

shunt-surgery-right-heart-catheterization-vascular-morphometry-rat

Research

JoVE Journal

Medicine

16.7K 次观看.  University Medical Center Groningen, University of Groningen. This protocol describes a surgical procedure to create a model for flow-induced pulmonary arterial hypertension (PAH) in rats and the procedures to analyze the principle hemodynamic and histological end-points in this model.

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

Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC twin pregnancies develop complications arising from the shared placenta and associated vascular connections1. Severe twin-to-twin syndrome (TTTS) is reported to account for approximately 20% of these complications2,3. Inter-twin vascular connections occur in almost all MC placentas and are related to the prognosis and outcome of these high-risk twin pregnancies. The number, size and type of connections have been implicated in the development of TTTS and other MC twin conditions. Three types of inter-twin vascular connections occur: 1) artery to vein connections (AVs) in which a branch artery carrying deoxygenated blood from one twin courses along the fetal surface of the placenta and dives into a placental cotyledon. Blood flows via a deep intraparenchymal capillary network into a draining vein that emerges at the fetal surface of the placenta and brings oxygenated blood toward the other twin. There is unidirectional flow from the twin supplying the afferent artery toward the twin receiving the efferent vein; 2) artery to artery connections (AAs) in which a branch artery from each twin meets directly on the superficial placental surface resulting in a vessel with pulsatile bidirectional flow, and 3) vein to vein connections (VVs) in which a branch vein from each twin meets directly on the superficial placental surface allowing low pressure bidirectional flow. In utero obstetric sonography with targeted Doppler interrogation has been used to identify the presence of AV and AA connections4. Prenatally detected AAs that have been confirmed by postnatal placental injection studies have been shown to be associated with an improved prognosis for both twins5. Furthermore, fetoscopic laser ablation of inter-twin vascular connections on the fetal surface of the shared placenta is now the preferred treatment for early, severe TTTS. 
Postnatal placental injection studies provide a valuable method to confirm the accuracy of prenatal Doppler ultrasound findings and the efficacy of fetal laser therapy6. Using colored dyes separately hand-injected into the arterial and venous circulations of each twin, the technique highlights and delineates AVs, AAs, and VVs. This definitive demonstration of MC placental vascular anatomy may then be correlated with Doppler ultrasound findings and neonatal outcome to enhance our understanding of the pathophysiology of MC twinning and its sequelae. Here we demonstrate our placental injection technique.

Vascular mapping of monochorionic (MC) twin placentas after birth provides a means for detailed demonstration of vascular connections between the twins&#x2019; circulations. 
Imbalance of these connections is thought to play a pivotal role in the development of complications of MC twinning including twin-to-twin transfusion syndrome.

导丝辅助导管和彩色染料注射的双人单绒毛膜胎盘血管映射

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC ...

科研

医学

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

股动脉和静脉导管采血，药品监督管理局和意识的血压和心率测量

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

右心小鼠的超声心动图评价

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

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

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. Pulmonary Hypertension (PH) was induced in 3-week-old piglets by a progressive obstruction of the pulmonary vascular bed. A ligation of the left Pulmonary Artery (PA) was performed first through a mini-thoracotomy. Second, weekly embolizations of the right lower pulmonary lobe were done under fluoroscopic guidance with n-butyl-2-cyanoacrylate during 5 weeks. Mean Pulmonary Arterial Pressure (mPAP) measured by ritght heart catheterism, increased progressively, as well as Right Atrial pressure and Pulmonary Vascular Resistances (PVR) after 5 weeks compared to sham animals. Right Ventricular (RV) structural and functional remodeling were assessed by transthoracic echocardiography (RV diameters, RV wall thickness, RV systolic function). RV elastance and RV-pulmonary coupling were assessed by Pressure-Volume Loops (PVL) analysis with conductance method. Histologic study of the lung and the right ventricle were also performed. Molecular analyses on RV fresh tissues could be performed through repeated transcutaneous endomyocardial biopsies. Pulmonary microvascular disease in obstructed and unobstructed territories was studied from lung biopsies using molecular analyses and pathology. Furthermore, the reliability and the reproducibility was associated with a range of PH severity in animals. Most aspects of the human CTEPH disease were reproduced in this model, which allows new perspectives for the understanding of the underlying mechanisms (mitochondria, inflammation) and new therapeutic approaches (targeted, cellular or gene therapies) of the overloaded right ventricle but also pulmonary microvascular disease.

Chronic Thromboembolic Pulmonary Hypertension (CTEPH) and Right Ventricular (RV) dysfunction were induced in piglets by progressive obstruction of the pulmonary arteries. Consequences were remarkably similar to those observed in CTEPH patients. This animal model would be a very useful tool for pathophysiology and therapeutic experiments on CTEPH and RV failure.

慢性血栓栓塞性肺动脉高压和右心室功能的仔猪评估

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

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.

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.

肺动脉高压的啮齿动物模型的血流动力学特性

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

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

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The morphological and functional phenotyping of the right ventricle is of particular importance in the context of hemodynamic compromise in patients with ARHF. Here, we describe a method to induce ARHF in a previously described large animal model of chronic PH, and to phenotype, dynamically, right ventricular function using the gold standard method (i.e., pressure-volume PV loops) and with a non-invasive clinically available method (i.e., echocardiography). Chronic PH is first induced in pigs by left pulmonary artery ligation and right lower lobe embolism with biological glue once a week for 5 weeks. After 16 weeks, ARHF is induced by successive volume loading using saline followed by iterative pulmonary embolism until the ratio of the systolic pulmonary pressure over systemic pressure reaches 0.9 or until the systolic systemic pressure decreases below 90 mmHg. Hemodynamics are restored with dobutamine infusion (from 2.5 &#181;g/kg/min to 7.5 &#181;g/kg/min). PV-loops and echocardiography are performed during each condition. Each condition requires around 40 minutes for induction, hemodynamic stabilization and data acquisition. Out of 9 animals, 2 died immediately after pulmonary embolism and 7 completed the protocol, which illustrates the learning curve of the model. The model induced a 3-fold increase in mean pulmonary artery pressure. The PV-loop analysis showed that ventriculo-arterial coupling was preserved after volume loading, decreased after acute pulmonary embolism and was restored with dobutamine. Echocardiographic acquisitions allowed to quantify right ventricular parameters of morphology and function with good quality. We identified right ventricular ischemic lesions in the model. The model can be used to compare different treatments or to validate non-invasive parameters of right ventricular morphology and function in the context of ARHF.

We present a protocol to induce and phenotype an acute right heart failure in a large animal model with chronic pulmonary hypertension. This model can be used to test therapeutic interventions, to develop right heart metrics&#160;or to improve the understanding of acute right heart failure pathophysiology.

慢性血栓栓塞性肺动脉高压大型动物模型中急性右心衰竭的诱导和表型分析

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

The mechanism of mitral stenosis-induced pulmonary venous arterialization and group 2 pulmonary hypertension (PH) is unclear. There is no rodent model of group 2 PH, due to mitral stenosis (MS), to facilitate the investigation of disease mechanisms and potential therapeutic strategies. We present a novel rat model of pulmonary venous congestion-induced pulmonary venous arterialization and group 2 PH caused by left atrial stenosis (LAS). LAS is achieved by constricting the left atrium using a half-closed titanium clip. After the LAS surgery, a rat model with a transmitral inflow velocity greater than or equal to 2.0 m/s on echocardiography gradually develops pulmonary venous arterialization and group 2 PH over an 8- to 10-week period. In this protocol, we provide the step-by-step procedure of how to perform the LAS surgery. The presented LAS rat model mimics MS in humans and is useful for studying the underlying molecular mechanism of pulmonary venous arterialization and for the preclinical evaluation of therapies for group 2 PH.

Left atrial stenosis (LAS) is a novel surgical technique used for studying group 2 pulmonary hypertension (PH) and mechanisms underlying pulmonary venous arterialization. Here, we present a protocol to constrict the left atrium using a titanium clip to cause pulmonary venous arterialization and moderate PH in a rat.

左心房狭窄致大鼠肺静脉动脉化及第2组肺动脉高压

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

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. However, the evaluation of right ventricular pressure (RVP) and pulmonary artery pressure (PAP) remains technically challenging in mice. RVP and PAP are more difficult to measure than left ventricular pressure because of the anatomical differences between the left and right heart systems. In this paper, we describe a stable right heart hemodynamic measurement method and its validation using healthy and PAH mice. This method is based on open-chest surgery and mechanical ventilation support. It is a complicated procedure compared to closed chest procedures. While a well-trained surgeon is required for this surgery, the advantage of this procedure is that it can generate both RVP and PAP parameters at the same time, so it is a preferable procedure for the evaluation of PAH models.

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach.

小鼠右心室系统及缺氧诱发肺动脉高血压的侵入性血液动力学评估

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...

Morphological and Functional Assessment of the Right Ventricle Using 3D Echocardiography

Traditionally, it was believed that the right side of the heart has a minor role in circulation; however, more and more data suggest that right ventricular (RV) function has strong diagnostic and prognostic power in various cardiovascular disorders. Due to its complex morphology and function, assessment of the RV by conventional two-dimensional echocardiography is limited: the everyday clinical practice usually relies on simple linear dimensions and functional measures. Three-dimensional (3D) echocardiography overcame these limitations by providing volumetric quantification of the RV free of geometrical assumptions. Here, we offer a step-by-step guide to obtain and analyze 3D echocardiographic data of the RV using the leading commercially available software. We will quantify 3D RV volumes and ejection fraction. Several technical aspects may help to improve the quality of RV acquisition and analysis as well, which we present in a practical manner. We review the current opportunities and the limiting factors of this method and also highlight the potential applications of 3D RV assessment in current clinical practice.

Here, we provide a step-by-step acquisition and analysis protocol for the 3D volumetric assessment of the right ventricle, mainly focusing on the practical aspects that maximize the feasibility of this technique.

使用3D超声心动图对右心室进行形态学和功能评估

Traditionally, it was believed that the right side of the heart has a minor role in circulation; however, more and more data suggest that right ...

A Rodent Model of The Ross Operation: Syngeneic Pulmonary Artery Graft Implantation in A Systemic Position

The Ross operation for aortic valve disease has regained new interest due to its outstanding long-term results. Nonetheless, when employed as freestanding root replacement, the possible dilation of the pulmonary autograft and subsequent aortic regurgitation is described. Several animal models have been proposed. However, these are usually limited to ex-vivo models or in-vivo experiments with relatively expensive large animal models. In this study, we sought to establish a rodent model of pulmonary artery graft (PAG) implantation in a systemic position. A total of 39 adult Lewis rats were included. Immediately after euthanasia, the pulmonary root was harvested from a donor animal (n=17). Syngeneic recipient (n=17) and sham-operated (n=5) rats were sedated and ventilated. In the recipient group, the PAG was implanted with an end-to-end anastomosis in infra-renal abdominal aortic position. Sham-operated rats underwent only transection and re-anastomosis of the aorta. Animals were followed with serial ultrasound studies for two months and post-mortem histological analysis. The median PAG diameter in the native position was 3.20 mm (IQR=3.18-3.23). At follow-up, the median diameter of the PAG was 4.03 mm (IQR=3.74-4.13) at 1 week, 4.07 mm (IQR=3.80-4.28) at 1 month, and 4.27 mm (IQR=3.90-4.35) at 2 months (p&lt;0.01). Peak systolic velocity was 220.07 mm/s (IQR=210.43-246.41) at 1 week, 430.88 mm/s (IQR=375.28-495.56) at 1 month, and 373.68 mm/s (IQR=305.78-429.81) at 2 months (p=0.02) and did not differ from the sham-operated group at the end of the experiment (p=0.5). Histological analysis did not show any sign of endothelial thrombosis. This study showed that rodent models may allow for the evaluation of the long-term adaptation of the pulmonary root to a high-pressure system. A systemically placed syngeneic PAG implantation represents a simple and feasible platform for the development and evaluation of novel surgical techniques and drug therapies to further improve the outcomes of the Ross operation.

We demonstrate how to establish a murine model of pulmonary root implantation into the descending aorta to simulate the Ross procedure. This model enables the medium/long-term evaluation of pulmonary autograft remodeling in a systemic position, representing the basis of developing therapeutic strategies to promote its adaptation.

罗斯手术的啮齿动物模型:全身位置的同源肺动脉移植植入

The Ross operation for aortic valve disease has regained new interest due to its outstanding long-term results. Nonetheless, when employed as freestanding ...