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
1Beijing Key Laboratory of Pre-clinical Research and Evaluation for Cardiovascular Implant Materials, State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Centre for Cardiovascular Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College, 2Laboratory Animal Center,Peking University, 3Center of Cardiac Surgery for Adults, State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Centre for Cardiovascular Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College
Summary
在这里，我们提出一个协议，使用开胸手术方法对小鼠的右心室和肺动脉进行侵入性血液动力学评估。
Abstract
肺动脉高血压（PAH）是一种慢性和严重的心肺疾病。老鼠是一种流行的动物模型，用来模仿这种疾病。然而，在小鼠中，右心室压力 （RVP） 和肺动脉压力 （PAP） 的评估在技术上仍然具有挑战性。RVP 和 PAP 比左心室压力更难测量，因为左心和右心系统之间的解剖差异。本文介绍了一种稳定的右心血液动力学测量方法，以及使用健康和PAH小鼠进行的验证。该方法基于开胸手术和机械通气支持。与封闭的胸腔手术相比，这是一个复杂的程序。虽然这种手术需要训练有素的外科医生，但这个程序的优点是它可以同时生成RVP和PAP参数，所以这是评估PAH模型的一个可取的程序。
Introduction
肺动脉高血压（PAH）是一种慢性、严重的心肺疾病，由小肺动脉细胞增殖和纤维化引起肺动脉压力（PAP）和右心室压力（RVP）升高。1.肺动脉导管，也称为天鹅-甘兹导管2，通常用于RVP和PAP的临床监测。此外，无线PAP监测系统已临床上使用3，4，5。为了模拟小鼠研究的疾病，使用低氧环境来模拟PAH6的人类临床表现。在动物PAP的评估中，大型动物使用与人类受试者相同的技术，通过肺动脉导管进行监测相对容易，但大鼠和小鼠等小动物由于体型小而难以评估。使用超小尺寸1Fr导管7，可以测量小鼠右心室系统的造声法。文献8、9报道了一种测量小鼠RVP和PAP的方法，但该方法缺乏详细的描述。RVP 和 PAP 比左心室压力更具测量性，因为左心和右心系统之间存在解剖学差异。
为了在同一只小鼠中同时获得PAP和RVP参数，我们描述了一种基于开胸手术的右心血液动力学测量方法，其验证与健康和PAH小鼠，以及如何避免在复杂的开胸期间生成人工数据手术。虽然这种技术最好由训练有素的外科医生执行，但它的优点是能够在同一位小鼠中评估 PAP 和 RVP。
Protocol
动物规程经中国医学科学院福威医院动物护理使用委员会审查通过，北京协和医学院（NO.0000287）。实验动物按照中国动物福利的指导方针进行饲养和喂养。 注：8至12周大的雄性C57BL小鼠被安置在12小时暗/12小时光周期的环境中。PAH小鼠在氧气浓度为10%的情况下被安置4周，由氧气控制的缺氧室维持，以诱发肺高血压，对照小鼠在相同条件下被安置在室内空气（21%氧气）。肥胖?…
Representative Results
压力传感器导管通过25G指针膨胀的隧道插入右心室（图3A），获得典型的RVP波形（图3C）。导管不断调整，缓慢推进，与肺动脉保持同一轴，同时通过肺瓣膜（图3B）。当压力传感器成功插入肺动脉时，出现了典型的具有双分波槽的PAP波形（图3D）。为了避免…
Discussion
气管插管是开胸手术的第一步。对于小动物（如大鼠或小鼠）来说，气管插管的经典方法包括在气管上做一个T形切口，并将Y型气管直接插入气管。在实践中，我们发现这种方法在操作过程中并不容易。Y 型气管管对于小动物来说太大，与气管形成一个角度。因此，很难将管固定到位。此外，一旦插管在开胸手术过程中意外地从气道流出，它通常会导致动物死亡，因为失去机械通气支持。因此，我…
Disclosures
The authors have nothing to disclose.
Acknowledgements
本研究由北京协和医学院研究生教育教学改革项目（10023-2016-002-03）、福威医院青年基金（2018-F09）和北京临床前研究重点实验室主任基金资助。心血管植入材料评估 （2018-PT2-ZR05）。
Materials
| 2,2,2-Tribromoethanol | Sigma-Aldrich | T48402-5G | For anesthesia |
| Animal temperature controller | Physitemp Instruments, Inc. | TCAT-2LV | For temperature control |
| Dissection forceps | Fine Science Tools, Inc. | 11274-20 | For surgery |
| Gemini Cautery System | Gemini | GEM 5917 | For surgery |
| Intravenous catheter (22G) | BD angiocath | 381123 | For intubation |
| LabChart 7.3 | ADInstruments | For data analysis | |
| Light illumination system | Olympus | For surgery | |
| Mikro-Tip catheter | Millar Instruments, Houston, TX | SPR-1000 | For pressure measurement |
| Millar Pressure-Volume Systems | Millar Instruments, Houston, TX | MVPS-300 | For pressure measurement |
| O2 Controller and Hypoxia chamber | Biospherix | ProOx 110 | For chronic hypoxia |
| PowerLab Data Acquisition System | ADInstruments | PowerLab 16/30 | For data recording |
| Scissors | Fine Science Tools, Inc. | 14084-08 | For surgery |
| Small animal ventilator | Harvard Apparatus | Mini-Vent 845 | For surgery |
| Stereomicroscope | Olympus | SZ61 | For surgery |
| Surgery tape | 3M | For surgery | |
| Terg-a-zyme enzyme | Sigma-Aldrich | Z273287-1EA | For catheter cleaning |
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Cite This Article
Luo, F., Wang, X., Luo, X., Li, B., Zhu, D., Sun, H., Tang, Y. Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice. J. Vis. Exp. (152), e60090, doi:10.3791/60090 (2019).