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Summary
Abstract
Introduction
Protocol
Results
Discussion
Disclosures
Acknowledgements
Materials
References
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Medicine

En Rat modell för ventrikelflimmer och Resuscitation av konventionell Stängd-bröst Technique

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

Nära 360.000 personer i USA 1 och många fler världen över 2 lider en episod av plötsligt hjärtstopp varje år. Försök att återställa livet kräver inte bara att hjärtaktivitet återupprättas men att skador på vitala organ förhindras, minimeras, eller omvänd. Aktuella hjärt-lungräddning tekniker ger en initial återupplivning takt på ca 30%; emellertid är överlevnad till utskrivning från sjukhus endast 5% en. Myokarddysfunktion, neurologisk dysfunktion, systemisk inflammation, tillstötande sjukdomar, eller en kombination därav inträffar efter återupplivning står för den stora andelen patienter som dör trots initial avkastning på cirkulation. Således är större förståelse för de underliggande patofysiologi och nya återupplivningsmetoder snarast för att öka graden av initial återupplivning och efterföljande överlevnad med intakt organfunktion.

Djurlägels av hjärtstillestånd spelar en avgörande roll i utvecklingen av nya återupplivnings terapier genom att ge insikter om patofysiologin vid hjärtstopp och återupplivning och erbjuder praktiska sätt att konceptualisera och testa nya ingripanden innan de kan testas på människor 3. Modellen råtta av slutna bröstet hjärt-lungräddning (HLR) som beskrivs här har spelat en viktig roll. Modellen utvecklades 1988 av Irene von Planta – forskarassistent vid tidpunkten – och hennes medarbetare 4 i laboratoriet sen professor Max Harry Weil MD, Ph.D. vid University of Health Sciences (omdöpt Rosalind Franklin University of Medicine and Science 2004) och har i stor utsträckning använts inom återupplivning främst genom stipendiater i professor Weil och deras praktikanter.

Modellen simulerar en episod av plötsligt hjärtstopp med återupplivning försökt med konventionella CPR tekniker och omfattar därmed intagetjonen av ventrikelflimmer (VF) genom att leverera en elektrisk ström till det högra ventrikulära endokardiet och tillhandahållande av sluten bröstkorg HLR genom en pneumatiskt driven kolv enhet medan samtidigt levererar övertrycksventilation med syreanrikad gas. Uppsägning av VF åstadkommes genom transtorakal leverans av elektriska stötar. Den råttmodell skapar en balans mellan modeller som utvecklats i stora djur (t.ex. svin) och modeller som utvecklats i mindre djur (t.ex. möss) möjliggör utforskande av nya forskningskoncept i en väl standardiserat, reproducerbar och effektivt sätt med tillgång till en robust inventering av relevanta mätningar. Modellen är särskilt användbar i tidiga stadier av forskning för att undersöka nya koncept och undersöka effekterna av confounders innan studier på modeller större djur som är dyrare, men större translationell genomslag.

En Medline söka efter alla vetenskapliga artiklar rapporteringimilar råttmodell med VF som mekanismen för hjärtstillestånd och någon form av sluten bröstet återupplivning visar totalt 69 andra originalstudier med hjälp av modellen sedan den först publicerades 1988 4. Forskningsområdena är patofysiologiska aspekter av återupplivning 5-17, faktorer som påverkar utfall 18-30, roll farmakologiska interventioner undersöker kärlsammandragande preparat 31-43, buffertmedel 44, inotropa medel 45, agenter som syftar till hjärtinfarkt eller cerebral skydd 46-70, och även effekterna av mesenkymala stamceller 71-73.

Modellen och protokoll som beskrivs i den här artikeln för närvarande används vid Resuscitation institutet. Ändå finns det flera möjligheter att "skräddarsy" den modell som bygger på den kapacitet som finns tillgängliga för enskilda utredare och målen för studierna.

Protocol

OBS: Protokollet godkändes av Institutional Animal Care och användning kommittén vid Rosalind Franklin University of Medicine and Science. Alla förfaranden var enligt Guide för skötsel och användning av försöksdjur som publicerats av National Research Council. 1. Experimentell Setup och anestesi Utför kalibreringar av de olika signalerna kan fångas med hjälp av ett datainsamlingssystem (tryck, temperatur, kolvförskjutning, elektrokardiogram [EKG], kapnografi,…

Representative Results

Råttan modell som beskrivs här användes nyligen för att jämföra effekterna av två hämmare av sarcolemmal natriumväteväxlar isoform 1 (NHE-1) på hjärtmuskel och hemodynamiska funktion under bröstkompressioner och efter återupplivning 61. Det har tidigare rapporterats att NHE-1-hämmare minskar hjärtmuskel reperfusionsskada genom att begränsa natrium-inducerad cytosoliskt och mitokondrie kalcium överbelastning, och därmed hjälpa till att bevara vänsterkammar distensibility under bröstkompre…

Discussion

Kritiska steg i protokollet

Det finns kritiska stegen i protokollet. När behärskar, fortsätt att förbereda och protokoll som koncist nedan. Den kirurgiska preparatet är snabbt, avancera katetrar snabbt genom små snitt utlöser minimal eller ingen kärlkramper och positionera kateterspetsarna som avsett, följt av en framgångsrik trakealintubation efter en enda eller ett fåtal försök (s); alltså, slutföra förberedelserna i ≈ 90 min från den initiala pentobarbital dose…

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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