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Abstract
Introduction
Protocol
Results
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Medicine

室颤由传统的封闭胸技术大鼠模型和复苏

Published: April 26, 2015
doi:
10.3791/52413
, ,
1Resuscitation Institute,Rosalind Franklin University of Medicine and Science

Summary

This article describes a rat model of electrically-induced ventricular fibrillation and resuscitation by chest compression, ventilation, and delivery of electrical shocks that simulates an episode of sudden cardiac arrest and conventional cardiopulmonary resuscitation. The model enables gathering insights on the pathophysiology of cardiac arrest and exploration of new resuscitation strategies.

Abstract

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic components of cardiopulmonary resuscitation in humans is herein described. The model was developed in 1988 and has been used in approximately 70 peer-reviewed publications examining a myriad of resuscitation aspects including its physiology and pathophysiology, determinants of resuscitability, pharmacologic interventions, and even the effects of cell therapies. The model featured in this presentation includes: (1) vascular catheterization to measure aortic and right atrial pressures, to measure cardiac output by thermodilution, and to electrically induce ventricular fibrillation; and (2) tracheal intubation for positive pressure ventilation with oxygen enriched gas and assessment of the end-tidal CO2. A typical sequence of intervention entails: (1) electrical induction of ventricular fibrillation, (2) chest compression using a mechanical piston device concomitantly with positive pressure ventilation delivering oxygen-enriched gas, (3) electrical shocks to terminate ventricular fibrillation and reestablish cardiac activity, (4) assessment of post-resuscitation hemodynamic and metabolic function, and (5) assessment of survival and recovery of organ function. A robust inventory of measurements is available that includes – but is not limited to – hemodynamic, metabolic, and tissue measurements. The model has been highly effective in developing new resuscitation concepts and examining novel therapeutic interventions before their testing in larger and translationally more relevant animal models of cardiac arrest and resuscitation.

Introduction

接近36万个人在美国1和许多世界各地的2患心脏骤停每年一个小插曲。试图恢复生活不仅要求心脏活动重新建立但重要器官的损害预防，尽量减少或逆转。当前心肺复苏技术得到的约30％的初始复苏速率然而，出院生存率只有5％1。心肌功能障碍，神经功能障碍，全身炎症，并发疾病，或其出现复苏后账户是谁，尽管流通的初始收益死亡的患者大比例的组合。因此，迫切需要的基本病理生理学和新颖复苏的方法更好的理解，以增加初始复苏和随后的存活与完整器官功能的速率。

动物模式心脏骤停的LS提供见解心脏骤停和复苏的病理生理机制，并提供构思和测试新的干预措施，才可以在人类3进行测试切实可行的手段发挥新的复苏疗法的发展具有重要作用。这里所描述的关胸心肺复苏的大鼠模型（CPR）等方面发挥了重要作用。在后期教授马克斯·哈里·韦伊医学博士的实验室和她的合作者4 -该模型在1988年开发的艾琳·冯·足底-研究员当时在健康科学大学（更名为医学和科学的罗莎琳德·富兰克林大学在2004年），并已广泛应用于复苏的领域主要由威尔教授和他们的学员的同伴。

该模型模拟心脏骤停复苏与用传统CPR技术未遂的情节，因此包括INDUCT离子心室纤颤（VF）通过提供到右心室心内膜和提供封闭胸CPR的电流由一个气动驱动的活塞装置，同时伴随地输送正压通气与富氧气体。终止VF是由胸交付电击来完成。大鼠模型撞击在大型动物（ 例如 ，猪）和模型在较小的动物研制开发的模型之间的平衡（ 例如 ，小鼠），允许新的研究概念在良好标准化，重复性好，高效的方式访问探索健壮库存相关的测量。该模型是在研究，探索新概念和研究混杂因素的影响进行研究的大动物模型更昂贵过，但更大的影响平移初期阶段尤其有用。

一个Medline检索所有同行评议的文章作为报告有VF为心脏骤停的机制，某种形式的胸外复苏imilar大鼠模型共使用模型69个原始研究发现，因为它在1988年第4年首次出版，研究领域包括复苏5-17病理生理方面，影响的结果18-30因素，药物干预研究血管加压剂31-43，缓冲剂的作用44，正性肌力药45，代理旨在心肌或脑保护46-70，并且还间质干细胞71-73的影响。

在这篇文章中描述的模型和协议，目前正在使用的复苏研究所。然而，有多个机会来“定制”的基础上提供给个人的调查和研究的目标的能力模型。

Protocol

注：本协议在医学和科学的罗莎琳德·富兰克林大学被批准的机构动物护理和使用委员会。所有的程序均符合美国国家研究委员会公布实验动物与指南的护理和使用 。 1.实验装置和麻醉用数据采集系统（压力，温度，活塞位移，心电图[心电图]，二氧化碳浓度监测仪等 ）被捕捉的各种信号的执行校准。 消毒器械和导管（ 例如 ，在高压釜的?…

Representative Results

此处所描述的大鼠模型最近使用期间胸部按压和复苏后61来比较对心肌和血液动力学功能的肌膜钠-氢交换亚型1（NHE-1）的2抑制剂的影响。据此前报道，NHE-1抑制剂通过限制钠引起的胞浆和线粒体钙超载减轻心肌再灌注损伤，从而有助于在胸外按压保持左室扩张，减轻复苏后心功能不全12。在这项研究中，NHE-1抑制剂cariporide（1毫克/千克），它已被广泛地研究，在过去，是使用较新?…

Discussion

在协议中的关键步骤

有在该协议的关键步骤。当掌握了，准备和协议进行如下简洁如下所述。外科手术的准备是迅速的，迅速推进导管通过小切口触发最小或没有血管痉挛和定位所述导管尖端的意图，随后成功气管插管后的单一或几个尝试（多个）;因此，完成了制备在≈90分钟，从最初的戊巴比妥剂量诱导的VF带内的参考值的基线测量（ 表1）。 VF是在每?…

Disclosures

The authors have nothing to disclose.

Acknowledgements

The authors would like to acknowledge Dr. Wanchun Tang MD, MCCM, FCCP, FAHA and Jena Cahoon of the Weil Institute of Critical Care Medicine in Rancho Mirage, CA. for their contributions to the resuscitation protocol outline and for having helped train the rodent surgeon (LL). The preparation of this article was in part supported by a gift in memory of US Navy Retired SKC Robert W. Ply by Ms. Monica Ply for research in heart disease and Parkinson’s disease and by a discretionary fund from the Department of Medicine at Rosalind Franklin University of Medicine and Science.

Materials

Name of Material/ Equipment Company Catalog Number Comments/Description
Sodium pentobarbital Sigma Aldrich P3761 http://www.sigmaaldrich.com/catalog/product/sigma/p3761?lang=en&region=US
Rectal thermistor BIOPAC Systems, INC TSD202A http://www.biopac.com/fast-response-thermistor
Needle electrode biopolar concentric 25 mm TP BIOPAC Systems, INC EL451 http://www.biopac.com/needle-electrode-concentric-25mm
PE25 polyethylene tubing  Solomon Scientific BPE-T25 http://www.solsci.com/products/polyethylene-pe-tubing
26GA female luer stub adapter Access Technologies LSA-26 http://www.norfolkaccess.com/needles.html
Stopcocks with luer connections; 3-way; male lock, non-sterile Cole-Parmer UX-30600-02 http://www.coleparmer.com/Product/Large_bore_3_way
_male_lock_stopcocks
_10_pack_Non_sterile/EW-30600-23
TruWave disposable pressure transducer Edwards Lifesciences PX600I  http://www.edwards.com/products/pressuremonitoring/Pages/truwavemodels.aspx?truwave=1
Type-T thermocouple Physitemp Instruments IT-18 http://www.physitemp.com/products/probesandwire/flexprobes.html
Central venous pediatric catheter  Cook Medical  C-PUM-301J https://www.cookmedical.com/product/-/catalog/display?ds=cc_pum1lp_webds
Abbocath-T subclavian I.V. catheter (14g x 5 1/2") Hospira 453527 http://www.hospira.com/products_and_services/iv_sets/045350427
Novametrix Medical Systems, Infrared CO2 monitor Soma Technology, Inc. 7100 CO2SMO  http://www.somatechnology.com/MedicalProducts/novametrix_respironics_co2smo_
7100.asp
Harvard Model 683 small animal ventilator Harvard Apparatus 555282 http://www.harvardapparatus.com/webapp/wcs/stores/servlet/haisku2_10001_11051_44453_-1_
HAI_ProductDetail_N_37322_37323
Double-flexible tipped wire guides Cook Medical  C-DOC-15-40-0-2 https://www.cookmedical.com/product/-/catalog/display?ds=cc_doc_webds
High accuracy AC LVDT displacement sensor Omega Engineering LD320-25 http://www.omega.com/pptst/LD320.html
HeartStart XL defibrillator/monitor Phillips Medical Systems M4735A http://www.healthcare.philips.com/main/products/resuscitation/products/xl/
Graefe micro dissection forceps 4 inches Roboz  RS-5135 http://shopping.roboz.com/Surgical-Instrument-Online-Shopping?search=RS-5135
Graefe micro dissection forceps 4 inches with teeth Roboz  RS-5157 http://shopping.roboz.com/Surgical-Instrument-Online-Shopping?search=RS-5157
Extra fine micro dissection scissors 4 inches Roboz  RS-5882 http://shopping.roboz.com/micro-scissors-micro-forceps-groups/micro-dissecting-scissors/Micro-Dissecting-Scissors-4-Straight-Sharp-Sharp
Heiss tissue retractor Fine Science Tools  17011-10 http://www.finescience.com/Special-Pages/Products.aspx?ProductId=321&CategoryId=134&
lang=en-US
Crile curve tip hemostats Fine Science Tools  13005-14 http://www.finescience.com/Special-Pages/Products.aspx?ProductId=372
Visistat skin stapler  Teleflex Incorporated 528135 http://www.teleflexsurgicalcatalog.com/weck/products/9936
Braided silk suture, 3-0 Harvard Apparatus 517706 http://www.harvardapparatus.com/webapp/wcs/stores/servlet/haisku2_10001_11051_43051_-1_
HAI_ProductDetail_N_37916_37936
Betadine solution Butler Schein 3660 https://www.henryscheinvet.com/
Sterile saline, 250 ml bags Fisher 50-700-069 http://www.fishersci.com/ecomm/servlet/itemdetail?catnum=50700069&storeId=10652
Heparin sodium injection, USP Fresenius Kabi 504201 http://fkusa-products-catalog.com/files/assets/basic-html/page25.html
Loxicom (meloxicam) Butler Schein 045-321 https://www.henryscheinvet.com/
Thermodilution cardiac output computer for small animals N/A N/A Custom-developed at the Resuscitation Institute using National Instruments hardware and LabVIEW software
Analog-to-digital data acquisition and analysis system N/A N/A Custom-developed at the Resuscitation Institute using National Instruments hardware and LabVIEW software
Pneumatically-driven and electronically controlled piston device for chest compression in small animals N/A N/A Custom-developed at the Weil Institute of Critical Care Medicine
60 Hz alternating current generator N/A N/A Custom-developed at the Weil Institute of Critical Care Medicine
DOWNLOAD MATERIALS LIST

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Keywords: Rat ModelVentricular FibrillationCardiac ResuscitationClosed-chest TechniqueCardiopulmonary ResuscitationVascular CatheterizationTracheal IntubationPositive Pressure VentilationElectrical ShocksHemodynamic MeasurementsMetabolic MeasurementsResuscitation ConceptsTherapeutic InterventionsCardiac Arrest
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Lamoureux, L., Radhakrishnan, J., Gazmuri, R. J. A Rat Model of Ventricular Fibrillation and Resuscitation by Conventional Closed-chest Technique. J. Vis. Exp. (98), e52413, doi:10.3791/52413 (2015).
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