表征隔离，通风，和仪表小鼠肺灌注与脉动流
Summary
以下协议概述的隔离，通风和稽核小鼠肺组织措施稳定或搏动性肺血管压力 – 流的关系，以量化的血流量，气流，气道的变化和血管的变化，右心室后负荷的影响的过程。
Abstract
隔离，通风，允许稳定和脉动的肺血管压力 – 流的关系，对肺动脉流量，流速波形，气道压力和左心房压力独立控制测量仪表小鼠肺癌准备。基于多点，稳定的压力流量曲线计算肺血管阻力，肺血管阻抗是搏动性的压力 – 流的频率范围内获得的曲线计算。由于目前临床上公认的，阻抗比电阻右心室后负荷的一个优越的措施，因为它包含了血管顺应性的影响，这是不容忽视的，尤其是在肺循环。 “三个代表”重要指标的阻抗-零赫兹阻抗Z 0，特性阻抗Z C，和波反射ř W指数-提供洞察到远端动脉横截面积流量，近端动脉僵硬度和上下游的阻抗不匹配，分别为。在隔离所得的全部结果，通风和灌注肺是独立于交感神经系统的音，卷状态和麻醉的效果。我们已经用这种技术来量化肺栓塞和慢性缺氧电阻和阻抗的影响，并区分行动网站（即近端与远端）血管活性药物和使用 Z C的压力依赖的疾病。此外，当这些技术与基因工程菌对小鼠的肺部用于，分子水平上对肺血管的结构和功能缺陷的影响，可以决定的。
Protocol
在这个协议中，我们展示了一个孤立的，通风，灌注小鼠肺准备以前已使用量化肺栓塞和慢性缺氧对搏动性肺血管压力流量关系的影响（Tuchscherer，韦伯斯特，Chesler，2006年; Tuchscherer等，2007）。简言之，小鼠肺手术从周围组织中分离出，放置在一个加热室（IL – 1，哈佛大学器械，Holliston，马）和通风（通气控制模块（VCM）- R的定时器计数器模块（TCM），哈佛大学器械）。肺血管灌注加热的RPMI…
Discussion
在手术的关键步骤
这是关键，护理是采取切割时从肺部肋骨。必须充分暴露肺和不羁周围组织在通货膨胀，但不是在隔离的过程中损坏。可用于使用一个平面物体如镊子后端举行胸壁肺剪刀削减，以便有一个清晰的路径。另一个关键的一步是缝合肺动脉和主动脉周围的位置。使用钝直镊子将降低肺动脉穿刺的风险。手术过程中的最后关键的一步是插管的位置。如果套管以上的飞…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
这项研究是由国家卫生部授予R01HL086939（NCC）研究院的支持。
Materials
| Material Name | Tipo | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| 1 ml syringe | Fisher Sci | 14-829-10F | ||
| 10 ml syringe | Fisher Sci | 14-823-2A | ||
| 60 ml syringe | Fisher Sci | 13-689-8 | ||
| RPMI with GLN 6/PK | Fisher Sci | MT10040CV | ||
| Bottle Top Filters | Fisher Sci | 09-761-57 | ||
| Ficoll PM 70 | Sigma-Aldrich | F2878-100g | ||
| Heparin | Sigma-Aldrich | |||
| Y27632 | Sigma-Aldrich | Y0503 | ||
| Angled Ball Iris scissors | Fine Science Tools | 14109-09 | ||
| Vannas Spring Scissors – 4mm Blades | Fine Science Tools | 15018-10 | ||
| Fine Iris Scissors – straight | Fine Science Tools | 14106-09 | ||
| Dumont #5/45 Forceps | Fine Science Tools | 11251-35 | ||
| Dumont Medical Biology Forceps | Fine Science Tools | 11254-20 | ||
| Lauda E100 ECO-line 003 | VWR | Comparable to Lauda-Brinkmann E-103, 62400-922 | ||
| IL-1 Isolated perfused mouse lung system | Harvard Apparatus | 739904 | ||
| Blood Pressure Transducer P75 for PLUGSYS Module | Harvard Apparatus | 730020 | ||
| TS410 Flow Modules | Transonic | TS410 | ||
| ME 4 PXN Precision PXN Inline Flowsensors | Transonic | ME 4 PXN | ||
| Cole-Parmer Multi-Syringe Pumps | Cole-Parmer | EW-74900-20 | ||
| Nembutal 50MG/ML 20ML Vial | Amatheon |
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Tags
Isolated Mouse LungVentilated Mouse LungInstrumented Mouse LungPulsatile FlowPulmonary Vascular Pressure-flow RelationshipsPulmonary Arterial Flow RateFlow Rate WaveformAirway PressureLeft Atrial PressurePulmonary Vascular ResistanceMulti-point Pressure-flow CurvesPulmonary Vascular ImpedanceRight Ventricular AfterloadVascular CompliancePulmonary CirculationImpedance MetricsZero Hertz Impedance Z0Characteristic Impedance ZCIndex Of Wave Reflection RWDistal Arterial Cross-sectional AreaProximal Arterial StiffnessUpstream-downstream Impedance MismatchSympathetic Nervous System ToneVolume Status AnesthesiaPulmonary EmboliChronic HypoxiaVasoactive Agents
Citazione di questo articolo
Vanderpool, R. R., Chesler, N. C. Characterization of the Isolated, Ventilated, and Instrumented Mouse Lung Perfused with Pulsatile Flow. J. Vis. Exp. (50), e2690, doi:10.3791/2690 (2011).