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Introduction
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Medicine

従来のクローズド·チェスト法による心室細動と蘇生のラットモデル

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

36万米国1内の個人と、より多くの世界的に2に近い、毎年突然の心停止のエピソードに苦しむ。人生を復元​​しようとすると、心臓の活動が再確立することがなく、重要な器官への損傷が、防止最小化、または逆転されていることだけでなく、必要とする。現在の心肺蘇生法は、約30％の初期蘇生率をもたらす。しかし、生存退院はわずか5％1です。心筋機能障害、神経機能障害、全身性炎症、併発疾患、または循環の最初のリターンにもかかわらず、死亡する患者の大きな割合を蘇生後のアカウントを発生するそれらの組み合わせ。したがって、基礎となる病態生理学および新規蘇生のより深い理解のアプローチが緊急に初期蘇生無傷臓器機能とその後の生存率を増加させるために必要とされる。

動物のモード彼らは人間の3で試験することができる前に、心停止のLSは心停止と蘇生の病態生理学上の洞察を提供し、新たな介入を概念化し、テストするための実用的な手段を提供することで、新たな蘇生治療の開発において重要な役割を果たしている。ここで説明し、閉じた胸の心肺蘇生（CPR）のラットモデルは、重要な役割を果たしてきました。後半教授マックスハリーワイル医学博士の研究室で4を 、彼女の共同研究者-当時の研究員-モデルは、アイリーン·フォン·プランタによって1988年に開発されました健康科学大学（200​​4年の医学と科学の名前を変更したロザリンド·フランクリン大学）で、かつ広範囲に教授ワイルとその研修生の仲間たちによって主に蘇生の分野で使用されてきた。

モデルは、従来のCPR技術によって試み蘇生で突然の心停止のエピソードをシミュレートし、したがって、入会さを含み付随して酸素富化ガスで陽圧換気を提供しながら、空気圧で駆動されるピストン装置による右心室心内膜と、閉じた胸のCPRの提供への電流を提供することにより、心室細動（VF）のイオン。 VFの終了は、電気ショックの経胸壁配信することによって達成される。ラットモデルは、堅牢へのアクセス権を持つ、よく標準化された再現可能、かつ効率的な方法で新しい研究概念の探査を可能に小さい動物（ 例えば 、マウス）で開発された大型動物（ 例えば 、豚）とモデルで開発されたモデルとの間のバランスを打つ適切な測定の目録。モデルは、新しい概念を探求し、より高価な、しかしより大きな翻訳への影響であるより大きな動物モデルで試験を実施する前に、交絡因子の影響を調べるために、研究の初期段階で、特に有用である。

すべての査読論文として報告するためのMedlineの検索それが最初に1988年4年に出版されたので、心停止のメカニズムと、閉じた胸の蘇生のいくつかのフォームとしてVFを持つimilarラットモデルは、モデルを使用して69追加のオリジナルの研究の合計を明らかにした。研究分野は、蘇生5-17の病態生理学的な側面が含まれ、成果18-30に影響を与える因子、血管収縮剤31-43、緩衝剤を検査薬理学的介入の役割44、強心薬45、心筋梗塞または脳保護を46から70に向けた薬剤とともに、間葉系幹細胞が71~73の効果。

この資料に記載されたモデルとプロトコルは、現在、蘇生研究所で使用されています。しかし、個々の研究者との研究の目標に利用可能な機能に基づいてモデルを「カスタマイズ」するために、複数の機会があります。

Protocol

注：プロトコルは、医学と科学のロザリンド·フランクリン大学の施設内動物管理使用委員会によって承認された。すべての手順は、米国学術研究会議によって公開実験動物の管理と使用に関する指針に従った。 1。実験のセットアップと麻酔データ収集システム（圧力、温度、ピストン変位、心電図[ECG]、カプノグラフィなど ） を用いて捕?…

Representative Results

ここで説明するラットモデルは、最近、胸部圧迫および蘇生後61中の心筋血行動態機能上の筋細胞のナトリウム-水素交換体アイソフォーム1（NHE-1）の2つの阻害剤の効果を比較した。これは、以前にNHE-1阻害剤はナトリウムによって誘発される細胞質ゾルおよびミトコンドリアのカルシウム過負荷を制限することによって、心筋再潅流傷害を減衰し、それによって、胸部圧迫中に左心?…

Discussion

プロトコルにおける重要なステップ

プロトコルの重要なステップがあります。マスターした場合、準備とプロトコルは、簡潔に以下のように進みます。外科的準備は、単一または少数の試み（S）の後に成功した気管挿管に続いて、最小または無血管攣縮をトリガーし、意図したとおりにカテーテル先端を配置する小さな切開を通じて急速にカテーテルを進め、迅…

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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