SR is supported by NIH K08HL114643, Gilead Research Scholars in Pulmonary Arterial Hypertension and a Burroughs Wellcome Fund Career Award for Medical Scientists.

中描述的所有程序遵循医学杜克大学医学院的动物护理准则。 1.在此之前启动程序注意:任何动物程序之前，确保已经获得适当的机构的许可。如同所有的程序，使用适当的止痛药，以确保没有任何动物的痛苦。 <ol><li>与肝素的无菌盐水（100单位/毫升）冲洗导管，以确保通畅。标记从导管等效于长度从心脏到颈部的中部的前端的点（约4cm老鼠和2厘米只小鼠）。 </li><li>麻醉小鼠或大鼠。麻醉剂的选择包括异氟烷（诱导3-4％，维持1.5％与100％的氧气的混合），氯胺酮/赛拉嗪（80-120 / 10毫克/千克）和戊巴比妥（40-80毫克/千克）6。 <ol><li>例如，用氯胺酮:赛拉嗪（80-120毫克/公斤:10-16毫克/千克IP进行小鼠和80-100毫克/公斤:5-10米克/千克IP进行鼠），以单剂量持续麻醉20-50分钟。超声心动图，麻醉小鼠或大鼠用异氟烷（诱导3-4％和维护1.5％）。按捏在啮齿动物手术区，以确认取款反射消失评估麻醉深度。使用于眼部兽医药膏，以防止干燥时的麻醉下。 注意:不同的麻醉剂可用于获得具有正确的使用和优化可靠的结果（综述别处6）。我们的导尿偏好是使用氯胺酮:甲苯噻嗪。过量用氯胺酮/赛拉嗪可能深刻地降低心脏速率和心功能，所以它是保持适当的温度和呼吸控制的关键。为了保持心脏小鼠率（&gt; 400 /分），我们经常进行双边迷走神经切断。氯胺酮/赛拉嗪的此处的量一般将最后20-30分钟，这是足以执行任一开闭胸心脏导管插入术，随后通过安乐死动物。 </li></ol></li><li>制备大鼠/小鼠的手术过程（ 图1）。 <ol><li>剃须从胸部皮毛（允许超声心动图），并从手术区，在右边的脖子上。 </li><li>从内向外的优碘的中心的圆形扫擦洗剃手术区域，接着用70％的酒精棉签清洁。 </li><li>将动物在手术平台下的一个暖垫。设定加热水平以维持体温的37-37.5℃。监测体温用直肠探针。低温可导致显著心动过速显著心动过缓和热疗结果。 </li></ol></li></ol> 2.超声心动图注:在其他地方7中描述的啮齿动物超声心动图的完整描述。为鼠标，前麻醉，可在清醒的，手动抑制动物获得的图像。对于大鼠，麻醉优选为大鼠之前心动过大，在清醒时手动抑制）。 <ol><li>胸骨旁长轴（PLAX）查看。 <ol><li>放置动物在平台上的仰卧姿势或手动抑制它。 </li><li>选择B模式投射二维实时图像。 </li><li>对准超声换能器40兆赫的鼠标或25兆赫的对大鼠的左胸骨旁线的频率，并然后旋转传感器逆时针30°与探针指示器在尾方向指向（5 11点线位置） 。角传感器略（沿同一断层平面中的换能器的短轴摇动），以获得在屏幕的中心的完整的低压室视图。 </li><li>找到并查看这些解剖结构（ 图2A）:左心室（LV）的内腔;室间隔（IVS）;右心室（RV）的管腔;升主动脉（AO）;和左心房（LA）。 </li> &lt;李&gt;切换到M模式下，一旦上述这些结构清晰可见。将通过利用AO作为参考点LV流明的最宽部分的指示线，也使左室腔（ 图2B）中央的聚焦深度谎言。通过改变传感器的角度和获取M模式下测量使RV的类似测量。 </li><li>用电影商店创建视频循环录制离线测量（LV腔尺寸，FS和左室壁厚度）的数据。 </li><li>通过将PW光标在主动脉和记录（ 图2C）取得在脉冲多普勒模式主动脉流出的多普勒跟踪。 </li></ol></li><li>胸骨旁短轴观（PSAX）在主动脉的水平。 <ol><li>切换到B模式。 </li><li>从胸骨旁长轴视图振子90°顺时针转动，得到胸骨旁短轴观（ 图3）。移动和角度传感器朝头盖骨为IDentify主动脉瓣横断面图。 </li><li>识别右室流出道（RVOT）作为定位于主动脉右上角月牙形结构，肺动脉瓣叶和肺动脉继续。 </li><li>握住手工同一位置稳定。切换到脉冲多普勒模式。 注意:用于保持啮齿动物和探针可以用于最小化换能器位置的移动和变形例的站台上。 </li><li>将样品体积近端肺动脉瓣的电平中的右心室流出道的中心，然后通过容器（ 图3B）平行于血液流动方向的光标定位。 注意:对采样角度调整到血液流动的方向，或使用超声软件以校正角度的变化是很重要的。未经校正，最大角度到容器为30°，其对应于速度的〜15％低估。 </li><li>形容词乌斯根据需要，得到"良好"的多普勒包络，它具有白色边框和暗中空内指示的层的血流（ 图3C）的比例（血流的速度）。记录多普勒跟踪。 注:一个"坏"多普勒信封不能容纳足够的白色边框，黑窟窿。 </li><li>如果在这个节骨眼上不进行导尿，允许如果使用麻醉啮齿动物恢复。直到它已经恢复了足够的意识，以保持胸骨斜卧，不交回公司的其他动物，直到其完全恢复，不要离开啮齿动物无人值守。如果进行导尿，请进入第3。 </li></ol></li></ol> 3.右心导管<ol><li>对于RV压力测量封闭胸方法<ol><li>建立: <ol><li>连接压力传感器输入通道1.在软件，设置通道1的压力和陈荫罴埃尔2心脏速率。 </li><li>到单位转换为毫米汞柱，记录基线轨迹，使用压力表手动（如果使用一个血压换能器和PE管）执行压力校准。然后执行下通道1单位换算。 </li><li>要设置心脏速率，关闭通道的输入2.选择下路2循环测量和选择源和速率测量通道1。 </li></ol></li><li>将鼠标/大鼠与深度的焦点和5倍的倍率解剖显微镜下。 </li><li>切割从下颌骨到胸骨（ 图1）的皮肤。放置一对拉钩到切口的每一侧以充分暴露出颈部区域。 </li><li>说白了解剖分离唾液腺，露出右颈外静脉用细钝尖镊子（ 图4A，B）。 </li><li>小心隔离从周围结缔组织右颈外静脉。 </li> &lt;li&gt;将丝线缝合两片（4-0老鼠; 6-0小鼠）右颈外静脉下方，结扎远端静脉（接近下颌骨越好），再扎一个松散的结近端（ 图4C）。 </li><li>使用虹膜剪，使一个小"缺口"（切）近端向远端打成结。 </li><li>握住镊子导管插入导管进入静脉切断，然后拧紧近端结。 注意:我们通常使用小鼠和聚乙烯（PE）-10油管（〜2星期五大小）的PE-50（〜3星期五大小）老鼠，其通过一个31 G或21g的针连接到常规压力传感器和校准。标记用标记在导管在一个长度大致对应于尖端放置在右心室。作为替代聚乙烯管材，可以使用微压计导管。轻轻拉动远端结可以帮忙介绍导管。 </li><li>轻轻推导管进入右心脏和监控根据标记的进​​步的深度。监控软件来验证导管位置，并确定RV压力压迹（ 图5）。 </li><li>保持导管动并收集数据（切换数据旁边的开始按钮记录）2分钟。 </li><li>继续样品采集（第四节）。 </li></ol></li><li>开胸方法对RV PV回路分析。 注意:不能以封闭胸腔方法由于电导导管，它不会从SVC传递到RA的刚度来执行右心室的光伏环分析。市售电导导管是专为LV光伏循环分析。 <ol><li>在设置通道1电导软件;通道2的压力;和信道3的心脏速率。 </li><li>插管与地下16聚四氟乙烯管大鼠和管连接到一个机械通气。计算并通过后续的设置通气参数为小鼠或大鼠ING公式6:潮气量（V T，毫升）= 6.2×M的1.01（M =动物质量，kg）;呼吸率（RR，分-1）= 53.5×M的-0.26（ 图6A）。 </li><li>蔓延70％乙醇到皮毛减少到手术区域毛皮的传播。 </li><li>让下面的xyphoid过程中的切口和双边剖析朝向侧面剪刀皮肤。 </li><li>穿过腹壁切割并通过沿隔膜双边夹层打开腹腔。 </li><li>打开隔膜暴露心脏的顶点和双边切肋骨（ 图6A）。通过喷雾盐水到使用注射器的胸椎和腹腔防止蒸发和组织的干燥。 注:我们通常用解剖剪打开腹腔和肋骨。出血通常是不显著的，但如果有出血，可以用电灼。 </li><li>仔细分离日从周围的结缔组织ê下腔静脉（IVC）。 </li><li>将一块丝线缝合（老鼠4-0; 6-0小鼠）周围的下腔静脉，再扎一个松散的结（或线程通过地下16聚四氟乙烯管缝合）（ 图6B）。 </li><li>穿刺平行于右室游离壁27-30号针头顶端RV游离壁并取出针。要小心，不要推针超过4毫米。 注意:可替换地，一小片的PE-60管，可用于指导电导导管穿刺到RV顶点。 </li><li>通过缠绕在心尖RV游离壁，直到所有电极是心室（ 图6C）内的刺插入电导导管末端。 </li><li>监控软件中的压力容积环，然后调整导管获得一致形循环，即不表现出显著的呼吸变化（ 图7B，C）的位置。 </li><li>记录基线PV循环（切换数据旁边的启动按钮录音）至少10秒获得了多项光伏循环。 </li><li>拉周围放置IVC缝线以改变预加载，并记录在PV环。分析离线数据并导出右心室收缩功能（ 图7D）的各种参数。此分析先前已8所述。 注:IVC可以备选地通过镊子闭塞。监控RV压迹，以确认预紧力的降低。 </li><li>如先前所描述，以允许从电导单位体积单位6转换执行盐水和试管校准。 </li><li>记录数据后，轻轻拉出导管和导管的末端放置立即与盐水的水浴中。在完成，清洁每制造商的说明在导管。 </li></ol></li></ol> 4.收集心脏和肺标本注意:这里的程序重新描述为终端，动物必须要么闭环或开胸右心脏导管插入后安乐死。 <ol><li>通过打开胸廓（双侧开胸）安乐死的小鼠如果使用封闭胸部的方式，血管扩张，或通过麻醉过量后关闭呼吸器。 注意:不建议颈椎脱位。 </li><li>为了进行肺灌注通胀，通胀管路连接到设置膨胀为20的水柱压力的肺部ringstand（但不要打开阀门尚未膨胀肺部）。 </li><li>说白了，从周围的肌肉和结缔组织解剖气管。 </li><li>将一块丝线缝合（老鼠4-0; 6-0小鼠）周围的气管，再扎一个松散的结。 </li><li>轻轻按下头部伸展气管，并进行切割（周长的70％）接近下颌骨。 </li><li>保持温柔的伸展并插入气管套管（20克的小鼠或大鼠为16 G）。固定用缝合插管。套管连接到通胀管和领带绕套管缝合，防止固定剂的回流。 </li><li>通过使用10毫升注射器刺伤右心室游离壁并朝肺动脉注入冲洗用PBS肺部。尼克一旦肺部开始变白左心房。 </li><li>通过在主动脉根部切割收获心脏。 </li><li>使用夹住一只蚊子止血肺的右下肺叶切右下肺叶。放置块放入离心管和捕捉在液氮中冷冻。 </li><li>膨胀，用10％缓冲的中性福尔马林肺5分钟并取出气管插管随后通过结扎气管。 </li><li>解剖肺出胸的，并用10％缓冲的中性福尔马林固定。 注意:可替换地，膨胀与最佳切割介质的肺（OCT中，1:1稀释，用PBS），并在未稀释十月冻结冰冻切片以后制备。 </li><li>小心分离从心室心房和通过解剖沿着室间隔隔离右心室游离壁。 </li><li>称量RV和LV +室间隔（LV + S）来计算富尔顿指数（RV / LV + S）9，它量化RV肥大的程度。 注:TheFulton指数PH不同型号而异。鼠10:控制，0.28±0.01;缺氧诱导，0.57±0.02; MCT处理，0.51±0.03。 C57BL6 / J小鼠11:控制，0.26±0.01; SuHx（14天），0.40±0.02; SuHx（21天），0.43±0.01; SuHx（28天），0.44±0.03。 </li><li>快冷冻在​​液氮中的休旅车和LV + S或在10％缓冲的中性福尔马林固定。 </li></ol>

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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=Bisoprolol+delays+progression+towards+right+heart+failure+in+experimental+pulmonary+hypertension.">Bisoprolol delays progression towards right heart failure in experimental pulmonary hypertension.</a> Circ Heart Fail. 5, 97-105 (2012).</li><li>de Man, F. 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=Dysregulated+renin-angiotensin-aldosterone+system+contributes+to+pulmonary+arterial+hypertension.">Dysregulated renin-angiotensin-aldosterone system contributes to pulmonary arterial hypertension.</a> Am J Respir Crit Care Med. 186, 780-789 (2012).</li><li>Pritts, C. D., Pearl, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthesia+for+patients+with+pulmonary+hypertension.">Anesthesia for patients with pulmonary hypertension.</a> Curr Opin Anaesthesiol. 23, 411-416 (2010).</li><li>Paulin, 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=A+miR-208-Mef2+Axis+Drives+the+Decompensation+of+Right+Ventricular+Function+in+Pulmonary+Hypertension.">A miR-208-Mef2 Axis Drives the Decompensation of Right Ventricular Function in Pulmonary Hypertension.</a> Circ Res. 116, 56-69 (2015).</li><li>Brittain, E., Penner, N. L., West, J., Hemnes, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Echocardiographic+assessment+of+the+right+heart+in+mice.">Echocardiographic assessment of the right heart in mice.</a> J Vis Exp. , (2013).</li><li>Cheng, H. 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=Assessment+of+right+ventricular+structure+and+function+in+mouse+model+of+pulmonary+artery+constriction+by+transthoracic+echocardiography.">Assessment of right ventricular structure and function in mouse model of pulmonary artery constriction by transthoracic echocardiography.</a> J Vis Exp. , e51041 (2014).</li></ol>

The authors have nothing to disclose.

The protocols outlined here describe a comprehensive characterization of hemodynamics and right ventricular function in rodent models of pulmonary hypertension. While right heart catheterization as described here is a terminal procedure, the mortality associated with echocardiography is minimal, which allows for screening and follow-up of disease progression. However, similar to patients with PH having markedly increased mortality with anesthesia17, in our experience, rats with severe PH do not tolerate anesthesia as well, due to decompensated right heart failure, decreased RV preload, and subsequently hypoxia-induced pulmonary vascular constriction. Therefore, slow induction and closely monitoring respiration are essential. 
The video and illustrations demonstrate what our laboratory has found to be successful with regards to these procedures, although alternative approaches can also be utilized (reviewed in Pacher et al.6). In PH research, these techniques are required in order to properly characterize different models of disease and the effects of genetic or pharmacologic manipulations on them. These methods each have their inherent limitations: for echocardiography, views of the right ventricle are limited and are difficult to quantify; for closed-chest catheterization, only an estimate of PA pressure is provided by an RVSP and an assessment of RV function is limited to dP/dt; for open-chest catheterization, the thorax must be opened, which can have effects on venous return and right heart function. 
Experience with these procedures is required to obtain reproducible and consistent results and there are a few critical steps where care is required. Determination of parameters from echocardiography requires standardized views, as small changes in probe angle can result in very different distance and Doppler measurements. During catheterization, introduction of a curve into the catheter can facilitate entry into the right ventricle; with polyethylene tubing, this can be done by heating the tubing gently; for micromanometer catheters, there are specialized catheters designed with a curve for this purpose. Dissection of the RV from the LV+S requires carefully en bloc harvest of the heart and the separation of the ventricles from the atria. As in any experiment, careful attention to detail with all of these procedures is critical for obtaining useful data. It is also important to be aware that animal respiration can affect the signal recording for pressure and PV loops. Identification and quantification of good and uniform pressure traces with consistently shaped loops are important in obtaining reliable data. 
This protocol will be of utility in research with animal models of PH and the right ventricle. Such studies include models of RV pressure overload (pulmonary artery banding) and right heart failure. These different experimental approaches allow quantification of pulmonary hypertension and RV function that are complementary. Cardiac catheterization has been used for the assessment of RV pressures, and recently investigators have reported success in traversing the pulmonic valve in the rat with micromanometer catheters designed for the mouse, allowing a direct measurement of PA pressures18. Open-chest PV loops allow a quantification of RV function; for example, in the rat MCT model, losartan significantly reduced RV afterload, restored ventricular-arterial coupling and improved RV diastolic function16. Noninvasive parameters derived from echocardiography have been used to estimate stroke volume, cardiac output and an estimate of PA pressures19,20. In total, these different techniques allow an assessment of the hemodynamic severity of pulmonary vascular disease and the RV response.

肺动脉高压（PAH）是炎性细胞浸润，平滑肌细胞增殖和内皮细胞凋亡相关联的肺血管的疾病。这些变化导致肺小动脉闭塞，随后导致右心室（RV）功能障碍和心脏衰竭。为了了解病理生理学的基本PAH PAH和右心衰竭，多个不同的模型，包括基因和药理学的模型，为研究这种疾病已经发展（综述别处1,2）。 这些模型的，最流行的是在小鼠低氧诱导（HX）PAH和野百合碱（MCT）和在大鼠SU5416缺氧（SuHx）模型。在小鼠HX模型，小鼠被暴露到4周缺氧（无论是常压或低压，对应于18000英尺的0.10的FiO2高度），与内侧扩散所得的发展，增加RV SYST奥利奇压力和RV肥大3的发展。 MCT在通过一个不明确的机制，然后导致PAH 4的发展在损伤肺的内皮细胞60毫克/公斤的结果的单剂量。 SU5416是血管内皮生长因子受体（VEGFR）1和2阻断剂的抑制剂，并用单次皮下注射60毫克/千克，随后通过暴露于慢性缺氧3周导致永久肺动脉高血压具有相似病理改变治疗到出现在人类疾病，闭塞性血管病变5的形成。在过去几年中，对肺动脉高血压几个转基因小鼠模型已被开发。这些包括敲除和骨形态发生蛋白受体2（BMPR2）的突变，如BMPR2基因突变在PAH的两个家族和特发性形式存在，血红素氧合酶1敲除和IL-6的过度表达（别处1,2-综述）。 PH这些不同的啮齿动物模型有不同程度的肺动脉高压，右心室肥厚和右心衰竭。而缺氧和各种转基因小鼠模型导致比任一大鼠模型1温和得多的PAH，它允许不同的基因突变和其相关的分子信号转导途径的测试。在MCT模型并导致严重的肺动脉高压，虽然MCT似乎是有毒的内皮细胞在多种组织4。该SuHx模型的特征是血管变化更加类似于在人类原发性肺动脉高压看出，虽然需要同时药理操纵和缺氧曝光。此外，在所有这些模型中，可能存在与PAH的发展相关的组织病理学变化，肺动脉压和右心室功能之间的断开。这是相对于人类疾病，其中通常有病理变化之间的比例关系，pulmon的严重性元高血压和右心衰竭的程度。因此，PH这些啮齿动物模型的综合表征是必需的，涉及到RV功能（通常通过超声心动图）的评估，血液动力学（心导管）和心脏​​组织病理学和肺（从组织收获）。 在这个协议中，我们描述了用于在大鼠和小鼠的PAH模型的血流动力学表征的基本技术。这些常规技术可以应用到右心室和肺血管的任何研究，并且不限于PAH的模型。通过超声心动图可视化RV是以大鼠相对简单，但在小鼠中更有挑战性，因为它们的大小和RV的复杂几何形状。此外，用于定量RV功能的一些替代物，比如TAPSE，肺动脉（PA）的加速时间和PA血流频谱开槽，不能很好的在人类验证，并用PU的相关评估只是弱lmonary高血压和血流动力学侵入RV功能。右心室血流动力学的确定是最好的一个封闭胸完成，以保持与灵感负胸内压力的影响，虽然开胸导尿使用阻抗导管允许压力容积（PV）的判定回路和更详细的血流动力学的表征。如同任何程序，开发经验与程序，是实验成功的关键。

<table border="0" cellpadding="0" cellspacing="0" width="983">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Vevo 2100 Imaging System (120V)&#160;</td>
			<td style="width:183px;">VisualSonics, inc.&#160;</td>
			<td style="width:191px;">VS-11945</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Vevo 2100 Imaging Station&#160;</td>
			<td style="width:183px;">VisualSonics, inc.&#160;</td>
			<td style="width:191px;"></td>
			<td></td>
		</tr>
		<tr height="27">
			<td height="27" style="height:27px;width:443px;">High-frequency Mechanical Transducers</td>
			<td style="width:183px;">VisualSonics, inc.&#160;</td>
			<td style="width:191px;">MS250, MS550D, MS400</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:443px;">Ultrasound Gel Parker&#160;</td>
			<td style="width:183px;">Laboratories Inc.&#160;</td>
			<td style="width:191px;">01-08</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PowerLab 4/35</td>
			<td>ADInstruments</td>
			<td>ML765</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Labchart 8</td>
			<td>ADInstruments</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">BP transducer with stopcock and cable</td>
			<td>ADInstruments</td>
			<td>MLT1199</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">BP transducer calibration kit</td>
			<td style="width:183px;">ADInstruments</td>
			<td style="width:191px;">MLA1052</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mikro-Tip Pressure Catheter for mouse</td>
			<td>Millar</td>
			<td>SPR-1000</td>
			<td>Alternative catheter available from Scisense FT111B (mouse) and FT211B (rat)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Mikro-Tip Pressure Catheter for rat</td>
			<td style="width:183px;">Millar</td>
			<td style="width:191px;">SPR-513</td>
			<td>Alternative catheter available from Scisense FT111B (mouse) and FT211B (rat)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Millar Mikro-Tip&#160;ultra-miniature PV loop catheter for mice</td>
			<td>Millar</td>
			<td>PVR-1035</td>
			<td>Alternative catheter available from Scisense FT112 (mouse)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Millar Mikro-Tip ultra miniature PV loop catheter for rats</td>
			<td style="width:183px;">Millar</td>
			<td style="width:191px;">SPR-869</td>
			<td>Alternative catheter available from Scisense FT112 (mouse)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Millar PV system MPVS-300&#160;</td>
			<td>Millar</td>
			<td>MPVS-300</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4-0 Silk Black Braid 100 Yard Spool</td>
			<td>Roboz Surgical</td>
			<td>SUT-15-2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">6-0 Silk Black Braid 100 Yard Spool</td>
			<td>Roboz Surgical</td>
			<td>SUT-14-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Iris Scissors, Delicate, Integra Miltex</td>
			<td>VWR</td>
			<td>21909-248</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">VWR Dissecting Scissors, Sharp/Blunt Tip</td>
			<td style="width:183px;">VWR</td>
			<td style="width:191px;">82027-588</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">VWR Delicate Scissors, 4 1/2&#34;</td>
			<td style="width:183px;">VWR</td>
			<td style="width:191px;">82027-582</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Two star Hemostats, Excelta</td>
			<td style="width:183px;">VWR</td>
			<td style="width:191px;">63042-090</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Neutral-buffered formalin</td>
			<td style="width:183px;">VWR</td>
			<td style="width:191px;">89370-094</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Crotaline</td>
			<td style="width:183px;">Sigma</td>
			<td style="width:191px;">C2401</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">SU5416</td>
			<td style="width:183px;">Tocris Biosciences</td>
			<td align="right" style="width:191px;">3037</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3.5X-45X Boom Stand Trinocular Zoom Stereo Microscope&#160;</td>
			<td>AmScope</td>
			<td>SM-3BX</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PE (Polyethylene Tubing)-10</td>
			<td>Braintree Scientific Inc</td>
			<td style="width:191px;">PE10 36 FT</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PE (Polyethylene Tubing)-50</td>
			<td>Braintree Scientific Inc</td>
			<td style="width:191px;">PE50 36 FT</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PE (Polyethylene Tubing)-60</td>
			<td style="width:183px;">Braintree Scientific Inc</td>
			<td style="width:191px;">PE60 36 FT</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tabletop Isoflurane Anesthesia Unit</td>
			<td>Kent Scientific</td>
			<td style="width:191px;">ACV-1205S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgisuite multi-functional surgical platform</td>
			<td>Kent Scientific</td>
			<td style="width:191px;">Surgisuite</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Retractor set</td>
			<td>Kent Scientific</td>
			<td>SURGI-5002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anesthesia induction chamber</td>
			<td>VetEquip</td>
			<td align="right" style="width:191px;">941443</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Anesthesia Gas filter canister</td>
			<td style="width:183px;">Kent Scientific</td>
			<td style="width:191px;">ACV-2001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:443px;">Rodent nose cone</td>
			<td style="width:183px;">VetEquip</td>
			<td align="right" style="width:191px;">921431</td>
			<td></td>
		</tr>
	</tbody>
</table>

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.

由于在啮齿类动物中右心脏导管插入术通常是不适用于纵向后续终端程序，超声心动图是筛查和随访12极好的无创性的选择。而在人类PAH肺动脉收缩压超声心动图通常是从三尖瓣关闭不全，通常是简单的顶视图中获得的衍生，是不是在啮齿类动物中获得可靠这样的观点，防止由多普勒肺动脉收缩压的估计。然而，在主动脉级PSAX视图可以容易地在啮齿类动物中，这使得记录和测量肺动脉多普勒跟踪，其形状已与肺动脉高压12的关联度可视化。超声心动图研究的代表性结果证明在图3中。在这个协议中，超声检查被蒙蔽动物r是治疗或程序eceived。结果进行线分析关闭。 右心脏导管检查和测量RVSP的，作为在无肺动脉狭窄的肺动脉收缩压的精度估计，在啮齿动物模型13,14为PAH的量化的黄金标准。在这个协议中，无论是对RV压力测量封闭胸的方法（ 图5）和开胸方式为PV RV循环分析（ 图6,7）给出15,16。闭胸方法的优点是比开胸方法侵入性更小，动物是一段较长时间6更稳定。此外，不需要使用这种方法正压通气也不是胸腔打开，保持与呼吸和胸内负压关联的正常右路充盈压。开放式胸方法允许使用电导导管和PV的决心循环，从它可以计算出右心室功能的重要参数。因此，这些方法是互补的，因为他们有不同的长处和短处。 在从小鼠HX模型所示的闭合胸数据时，RVSP在45毫米汞柱，具有显著肺动脉高压（ 图5）一致升高。在从正常大鼠中所示的开胸数据，所述RVSP是显著下，在27毫米汞柱（ 图7）。 X轴的相对体积单位（RVU）可以反应杯校准之后被转换为体积单位，随后盐水校准以除去电导的部件由于心脏壁6,8。这就使心脏功能的重要参数，如收缩（通常作为评估的收缩末期弹性，E ES），舒张功能（从舒张末期压力容积关系），动脉弹性（E A）和计算预紧-recruitable工作行程，CA其中lculations在别处6,8讨论。 <img alt="图1" src="/files/ftp_upload/53335/53335fig1.jpg" /> 图 1: 啮齿动物对过程的制备大鼠麻醉并在胸部和颈部剃毛。红色虚线表示将用于暴露颈外静脉的切口。黑线代表了锁骨胸骨。蓝色圆圈表示超声心动图探头位置。 <img alt="图2" src="/files/ftp_upload/53335/53335fig2.jpg" /> 图 2: 回波的观点不同解剖结构的这些代表性图像是从正常小鼠。 （ 一 ）胸骨旁长轴（PLAX）视图。 LA:左心房; LV:左心室内腔; IVS:室间隔; RV:右v的管腔entricle; AO:升主动脉（AO）。 （注:上PLAX不同成像取向可导致不同的影像的约定。）（B）的M型与LV收缩（LVS）的LV和舒张（LVD）直径，以及前部（AWT）和后壁厚度（PWT）指出。短轴缩短率的计算公式为（LVD-LVS）/ LVD。 （C）脉冲多普勒主动脉展示主动脉流出的信号。 <a href="https://www.jove.com/files/ftp_upload/53335/53335fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/53335/53335fig3.jpg" /> 图3:胸骨旁短轴（PSAX）和右室次这些代表性图像从使用MCT PAH大鼠。 （ 一 ）在右心室的中间水平PAP视图PSAX。 （B）在主动脉水平PSAX图。右室流出道:右心室流出TR法案。 PA:肺动脉。 AO:主动脉。 （C）脉冲多普勒模式。样品体积（黄线）被放置在右心室流出道近端肺动脉瓣水平的中心。 <a href="https://www.jove.com/files/ftp_upload/53335/53335fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/53335/53335fig4.jpg" /> 图4:对大鼠的导尿颈外静脉的曝光 （A）中从下颌骨到作出胸骨和一对拉钩的切口放置到切口的每一侧，以暴露颈区域。唾液是腺（SG）被覆盖在颈外静脉（EJ）。 （B），说白了解剖，分离唾液腺及周围结缔组织，充分调动右颈外静脉。 （C）将围绕右颈外静脉远端和近端4-0丝线缝合。用作压力导管（D）一种PE-50管插入右侧EJ。 SG:唾液腺; EJ:颈外静脉; DS:远端缝合; PS:近端缝合;导管:导管。 <img alt="图5" src="/files/ftp_upload/53335/53335fig5.jpg" /> 图5:右心脏导管手术中不同腔室波形低氧性肺动脉高压鼠标右心脏导管插入术过程中的压力变化代表性的痕迹。面板左，中，随着时间的推移正确显示压力变化（毫米汞柱）（秒）的上腔静脉（静脉），右心房（RA），右心室（RV）。 <img alt="图6" src="/files/ftp_upload/53335/53335fig6.jpg" /> 图6:RV导管置入开胸方法。 </气管插管后STRONG&gt;（A）查看，通过腹壁切割，打开隔膜暴露心脏的顶点和双边切肋骨。一块周围的IVC缝合的（B）的分离和放置.;和（C）通过右心室心尖游离壁的电导导管插入后。 <img alt="图7" src="/files/ftp_upload/53335/53335fig7.jpg" /> 图7:右心室压力容量环分析 （ 一 ）在软件演示传导通道（RVU -相对体积单位），RV压力（毫米汞柱）和心脏速率（BPM）。 7-11次平滑需要获得良好的信号。在容易出现在光伏循环是可变的呼吸的结果变化的区域中的电导导管（B）的布局。 （ 三 ）稳定PV环适当PLAC电导导管键相。 （D）PV代表家人对下腔静脉的压力缓解后循环。曲线此系列器件可收缩末期弹性的计算（E ES -心肌收缩的指标）和血管顺应性（E 一 -肺血管顺应性的措施）。 <a href="https://www.jove.com/files/ftp_upload/53335/53335fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension at JoVE.com

Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension

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 ...

hemodynamic-characterization-rodent-models-pulmonary-arterial

Nanyang Technological University

Research

JoVE Journal

Medicine

20.8K 次观看.  Duke University Medical Center. 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.

视频: Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension

Magnetic Resonance Imaging Quantification of Pulmonary Perfusion using Calibrated Arterial Spin Labeling

This demonstrates a MR imaging method to measure the spatial distribution of pulmonary blood flow in healthy subjects 
during normoxia (inspired O2, fraction (FIO2) = 0.21) hypoxia (FIO2 = 0.125), and hyperoxia 
(FIO2 = 1.00). In addition, the physiological responses of the subject are monitored in the MR scan environment. MR images 
were obtained on a 1.5 T GE MRI scanner during a breath hold from a sagittal slice in the right lung at functional residual capacity. An arterial 
spin labeling sequence (ASL-FAIRER) was used to measure the spatial distribution of pulmonary blood flow 1,2 and a multi-echo fast 
gradient echo (mGRE) sequence 3 was used to quantify the regional proton (i.e. H2O) density, allowing the quantification 
of density-normalized perfusion for each voxel (milliliters blood per minute per gram lung tissue). 
With a pneumatic switching valve and facemask equipped with a 2-way non-rebreathing valve, different oxygen concentrations 
were introduced to the subject in the MR scanner through the inspired gas tubing. A metabolic cart collected expiratory gas via expiratory tubing. Mixed expiratory O2 and CO2 concentrations, oxygen consumption, carbon dioxide production, respiratory exchange ratio, 
respiratory frequency and tidal volume were measured. Heart rate and oxygen saturation were monitored using pulse-oximetry. 
Data obtained from a normal subject showed that, as expected, heart rate was higher in hypoxia (60 bpm) than during normoxia (51) or hyperoxia (50) and the arterial oxygen saturation (SpO2) was reduced during hypoxia to 86%. Mean ventilation was 8.31 L/min BTPS during hypoxia, 7.04 L/min during normoxia, and 6.64 L/min during hyperoxia. Tidal volume was 0.76 L during hypoxia, 0.69 L during normoxia, and 0.67 L during hyperoxia. 
Representative quantified ASL data showed that the mean density normalized perfusion was 8.86 ml/min/g during hypoxia, 8.26 ml/min/g during normoxia and 8.46 ml/min/g during hyperoxia, respectively. In this subject, the relative dispersion4, an index of global heterogeneity, was increased in hypoxia (1.07 during hypoxia, 0.85 during normoxia, and 0.87 during hyperoxia) while the fractal dimension (Ds), another index of heterogeneity reflecting vascular branching structure, was unchanged (1.24 during hypoxia, 1.26 during normoxia, and 1.26 during hyperoxia). 
Overview. This protocol will demonstrate the acquisition of data to measure the distribution of pulmonary perfusion noninvasively under conditions of normoxia, hypoxia, and hyperoxia using a magnetic resonance imaging technique known as arterial spin labeling (ASL). 
Rationale: Measurement of pulmonary blood flow and lung proton density using MR technique offers high spatial resolution images which can be quantified and the ability to perform repeated measurements under several different physiological conditions. In human studies, PET, SPECT, and CT are commonly used as the alternative techniques. However, these techniques involve exposure to ionizing radiation, and thus are not suitable for repeated measurements in human subjects.

A MR imaging method to study the distribution of pulmonary blood flow under a variety of physiological conditions, in this case exposure to three different inspired oxygen concentrations: hypoxia, normoxia, and hyperoxia, is described. This technique utilizes human pulmonary physiology research techniques in an MR scanning environment.

使用校准动脉自旋标记磁共振肺灌注显像定量

This demonstrates a MR imaging method to measure the spatial distribution of pulmonary blood flow in healthy subjects 
during normoxia (inspired O2, ...

科研

医学

Increasing Pulmonary Artery Pulsatile Flow Improves Hypoxic Pulmonary Hypertension in Piglets

Pulmonary arterial hypertension (PAH) is a disease affecting distal pulmonary arteries (PA). These arteries are deformed, leading to right ventricular failure. Current treatments are limited. Physiologically, pulsatile blood flow is detrimental to the vasculature. In response to sustained pulsatile stress, vessels release nitric oxide (NO) to induce vasodilation for self-protection. Based on this observation, this study developed a protocol to assess whether an artificial pulmonary pulsatile blood flow could induce an NO-dependent decrease in pulmonary artery pressure. One group of piglets was exposed to chronic hypoxia for 3 weeks and compared to a control group of piglets. Once a week, the piglets underwent echocardiography to assess PAH severity. At the end of hypoxia exposure, the piglets were subjected to a pulsatile protocol using a pulsatile catheter. After being anesthetized and prepared for surgery, the jugular vein of the piglet was isolated and the catheter was introduced through the right atrium, the right ventricle and the pulmonary artery, under radioscopic control. Pulmonary artery pressure (PAP) was measured before (T0), immediately after (T1) and 30 min after (T2) the pulsatile protocol. It was demonstrated that this pulsatile protocol is a safe and efficient method of inducing a significant reduction in mean PAP via an NO-dependent mechanism. These data open up new avenues for the clinical management of PAH.

Pulmonary hypertension is associated with a significant reduction in pulmonary artery pulsatility, contractility and elasticity, contributing to an increase in pulmonary artery pressure and pulmonary resistance. Using a hypoxic piglet model, this study demonstrated that improving pulmonary artery plasticity using a newly developed pulsatile catheter improves hypoxic pulmonary hypertension.

增加肺动脉脉动流改善缺氧性肺动脉高压的仔猪

Pulmonary arterial hypertension (PAH) is a disease affecting distal pulmonary arteries (PA). These arteries are deformed, leading to right ventricular ...

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. ...

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.

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 ...

Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. Portal hypertension (PHT) due to cirrhosis is responsible for the most severe complications in patients with liver disease. The full picture of the portal hypertensive syndrome is characterized by increased portal pressure (PP) due to the increased intrahepatic vascular resistance (IHVR), hyperdynamic circulation, and increased splanchnic blood flow. Progressive splanchnic arterial vasodilation and increased cardiac output with elevated heart rate (HR) but low arterial pressure characterizes the portal hypertensive syndrome.
Novel therapies are currently being developed that aim to decrease PP by either targeting IHVR or increased splanchnic blood flow &#8212; but side effects on systemic hemodynamics may occur. Thus, a detailed characterization of portal venous, splanchnic, and systemic hemodynamic parameters, including measurement of PP, portal venous blood flow (PVBF), mesenteric arterial blood flow, mean arterial pressure (MAP), and HR is needed for preclinical evaluation of the efficacy of novel treatments for PHT. Our video article provides the reader with a structured protocol for performing invasive hemodynamic measurements in cirrhotic rats. In particular, we describe the catheterization of the femoral artery and the portal vein via an ileocolic vein and the measurement of portal venous and splanchnic blood flow via perivascular Doppler-ultrasound flow probes. Representative results of different rat models of PHT are shown.

Here we describe a detailed protocol for invasive measurements of hemodynamic parameters including portal pressure, splanchnic blood flow, and systemic hemodynamics in order to characterize the portal hypertensive syndrome in rats.

肝硬化大鼠门脉高压综合征的侵袭性血流动力学特征

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. ...

Invasive Hemodynamic Monitoring of Aortic and Pulmonary Artery Hemodynamics in a Large Animal Model of ARDS

One of the leading causes of morbidity and mortality in patients with heart failure is right ventricular (RV) dysfunction, especially if it is due to pulmonary hypertension. For a better understanding and treatment of this disease, precise hemodynamic monitoring of left and right ventricular parameters is important. For this reason, it is essential to establish experimental pig models of cardiac hemodynamics and measurements for research purpose. 
This article shows the induction of ARDS by using oleic acid (OA) and consequent right ventricular dysfunction, as well as the instrumentation of the pigs and the data acquisition process that is needed to assess hemodynamic parameters. To achieve right ventricular dysfunction, we used oleic acid (OA) to cause ARDS and accompanied this with pulmonary artery hypertension (PAH). With this model of PAH and consecutive right ventricular dysfunction, many hemodynamic parameters can be measured, and right ventricular volume load can be detected. 
All vital parameters, including respiratory rate (RR), heart rate (HR) and body temperature were recorded throughout the whole experiment. Hemodynamic parameters including femoral artery pressure (FAP), aortic pressure (AP), right ventricular pressure (peak systolic, end systolic and end diastolic right ventricular pressure), central venous pressure (CVP), pulmonary artery pressure (PAP) and left arterial pressure (LAP) were measured as well as perfusion parameters including ascending aortic flow (AAF) and pulmonary artery flow (PAF). Hemodynamic measurements were performed using transcardiopulmonary thermodilution to provide cardiac output (CO). Furthermore, the PiCCO2 system (Pulse Contour Cardiac Output System 2) was used to receive parameters such as stroke volume variance (SVV), pulse pressure variance (PPV), as well as extravascular lung water (EVLW) and global end-diastolic volume (GEDV). Our monitoring procedure is suitable for detecting right ventricular dysfunction and monitoring hemodynamic findings before and after volume administration.

We present a protocol of creating right ventricular dysfunction in a pig model by inducing ARDS. We demonstrate invasive monitoring of left and right ventricular cardiac output using flow probes around the aorta and the pulmonary artery, as well as blood pressure measurements in the aorta and pulmonary artery.

ards 大型动物模型主动脉和肺动脉血流动力学的侵袭血流动力学监测

One of the leading causes of morbidity and mortality in patients with heart failure is right ventricular (RV) dysfunction, especially if it is due to ...

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 ...

The Left Pneumonectomy Combined with Monocrotaline or Sugen as a Model of Pulmonary Hypertension in Rats

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats with the additional administration of monocrotaline (MCT) or SU5416 (Sugen). We also compare these two models to other PAH models commonly used in research. In the last few years, the focus of animal PAH models has moved towards studying the mechanism of angioproliferation of plexiform lesions, in which the role of increased pulmonary blood flow is considered as an important trigger in the development of severe pulmonary vascular remodeling. One of the most promising rodent models of increased pulmonary flow is the unilateral left pneumonectomy combined with a "second hit" of MCT or Sugen. The removal of the left lung leads to increased and turbulent pulmonary blood flow and vascular remodeling. Currently, there is no detailed procedure of the pneumonectomy surgery in rats. This article details a step-by-step protocol of the pneumonectomy surgical procedure and post-operative care in male Sprague-Dawley rats. Briefly, the animal is anesthetized and the chest is opened. Once the left pulmonary artery, pulmonary vein, and bronchus are visualized, they are ligated and the left lung is removed. The chest then closed and the animal recovered. Blood is forced to circulate only on the right lung. This increased vascular pressure leads to a progressive remodeling and occlusion of small pulmonary arteries. The second hit of MCT or Sugen is used one week post-surgery to induce endothelial dysfunction. The combination of increased blood flow in the lung and endothelial dysfunction produces severe PAH. The primary limitation of this procedure is that it requires general surgical skills.

The rodent left pneumonectomy is a valuable technique in pulmonary hypertension research. Here, we present a protocol to describe the rat pneumonectomy procedure and postoperative care to ensure minimal morbidity and mortality.

左肺切除术联合单氯联氨酸或苏根为大鼠肺动脉高压的模型

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats ...

Induction and Characterization of Pulmonary Hypertension in Mice using the Hypoxia/SU5416 Model

Pulmonary Hypertension (PH) is a pathophysiological condition, defined by a mean pulmonary arterial pressure exceeding 25 mm Hg at rest, as assessed by right heart&#160;catheterization. A broad spectrum of diseases can lead to PH, differing in their etiology, histopathology, clinical presentation, prognosis, and response to&#160;treatment. Despite significant progress in the last years, PH remains an uncured disease. Understanding the underlying mechanisms can pave the way for the development of new therapies. Animal models are important research tools to achieve this goal. Currently, there are several models available for recapitulating PH. This protocol describes a two-hit mouse PH model. The stimuli for PH development are hypoxia and the injection of SU5416, a vascular endothelial growth factor (VEGF) receptor antagonist. Three weeks after initiation of Hypoxia/SU5416, animals develop pulmonary vascular remodeling imitating the histopathological changes observed in human PH (predominantly Group 1). Vascular remodeling in the pulmonary circulation results in the remodeling of the right&#160;ventricle (RV). The procedures for measuring RV pressures (using the open chest method), the morphometrical analyses of the RV (by dissecting and weighing both cardiac ventricles) and the histological assessments of the remodeling (both pulmonary by assessing vascular remodeling and cardiac by assessing RV cardiomyocyte hypertrophy and fibrosis) are described in detail. The advantages of this protocol are the possibility of the application both in wild type and in genetically modified mice, the relatively easy and low-cost implementation, and the quick development of the disease of interest (3 weeks). Limitations of this method are that mice do not develop a severe phenotype and PH is reversible upon return to normoxia. Prevention, as well as therapy studies, can easily be implemented in this model, without the necessity of advanced skills (as opposed to surgical rodent models).

This protocol describes the induction of pulmonary hypertension (PH) in mice based on the exposure to hypoxia and the injection of a VEGF receptor antagonist. The animals develop PH and right ventricular (RV) hypertrophy 3 weeks after the initiation of the protocol. The functional and morphometrical characterization of the model is also presented.

使用缺氧/SU5416模型对小鼠的肺高血压进行诱导和特征

Pulmonary Hypertension (PH) is a pathophysiological condition, defined by a mean pulmonary arterial pressure exceeding 25 mm Hg at rest, as assessed by ...