JoVE Journal > Medicine
Summary
Abstract
Introduction
Protocol
Results
Discussion
Disclosures
Acknowledgements
Materials
References
Automatic Translation
Medicine

En Rat Modell av ventrikkelflimmer og Resuscitation av Konvensjonell Lukket brystet 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

Nær 360.000 personer i USA en og mange flere verden over to lider en episode av plutselig hjertestans hvert år. Forsøk på å gjenopprette livet krever ikke bare at hjerteaktivitet gjenopprettes, men det skader på vitale organer forebygges, minimert, eller reversert. Nåværende hjerte-lungeredning teknikker gi en innledende gjenoppliving hastighet på ca 30%; derimot, er overlevelsen til sykehuset bare 5% 1. Hjerteinfarkt dysfunksjon, nevrologisk dysfunksjon, systemisk inflammasjon, interkurrente sykdommer, eller en kombinasjon av disse forekommer post-lungeredning konto for den store andelen av pasienter som dør på tross av innledende retur av sirkulasjon. Dermed er større forståelse av de underliggende patofysiologi og romanen gjennoppliving tilnærminger sterkt behov for å øke frekvensen av initial gjenoppliving og påfølgende overlevelse med intakt organfunksjon.

Dyr models av hjertestans spille en avgjørende rolle i utviklingen av nye gjennoppliving terapi ved å gi innsikt i patofysiologien av hjertestans og lungeredning og tilbyr praktisk måte å conceptualize og teste nye intervensjoner før de kan bli testet på mennesker tre. Rottemodell for lukket kiste hjerte-lungeredning (HLR) er beskrevet her har spilt en viktig rolle. Modellen ble utviklet i 1988 av Irene von Planta – en stipendiat på den tiden – og hennes samarbeidspartnere 4 i laboratoriet av avdøde professor Max Harry Weil MD, Ph.D. ved University of Health Sciences (omdøpt Rosalind Franklin medisinske og vitenskapelige universitet i 2004), og har vært mye brukt innen gjenoppliving hovedsakelig av stipendiater av professor Weil og sine traineer.

Modellen simulerer en episode av plutselig hjertestans med gjenoppliving forsøkt av konvensjonelle HLR teknikker og dermed inkluderer Induction av ventrikkelflimmer (VF) ved å levere en elektrisk strøm til høyre ventrikkel endocardium og formidling av lukket kiste HLR av en pneumatisk drevet stempel enhet mens samtidig levere overtrykksventilasjon med oksygenberiket gass. Oppsigelse av VF oppnås ved transtorakal leveranser av elektriske støt. Rottemodellen treffer en balanse mellom modeller utviklet i store dyr (for eksempel svin) og modeller utviklet i mindre dyr (for eksempel mus) slik utforskning av nye forsknings konsepter i et godt standardisert, reproduserbar, og effektiv måte med tilgang til en robust beholdning av relevante målinger. Modellen er spesielt nyttig i tidlige stadier av forskning for å oppdage nye konsepter og undersøke effekten av confounders før gjennomføring av undersøkelser i større dyremodeller som er mer kostbart, men av større translasjonell innvirkning.

En Medline søk etter alle fagfellevurderte artikler rapportering somimilar rottemodell har VF som mekanismen av hjertestans og noen form for lukket kiste lungeredning avdekket totalt 69 flere originale studier ved hjelp av modellen siden det først ble utgitt i 1988 fire. De forskningsområder omfatter patofysiologiske aspekter ved gjenoppliving 5-17, faktorer som påvirker utfall 18-30, rollen av farmakologiske intervensjoner undersøke karkontraherende agenter 31-43, buffermidler 44, inotrope midler 45, agenter rettet mot hjerteinfarkt eller cerebral beskyttelse 46-70, og også effekten av mesenchymale stamceller 71-73.

Modellen og protokoll beskrevet i denne artikkelen blir nå brukt på Resuscitation Institute. Likevel, det er flere muligheter til å "tilpasse" modellen basert på egenskapene som er tilgjengelige for individuelle etterforskere og målene fra studiene.

Protocol

MERK: Protokollen ble godkjent av Institutional Animal Care og bruk komité ved Rosalind Franklin medisinske og vitenskapelige universitet. Alle prosedyrer var i samsvar med Guide for omsorg og bruk av forsøksdyr publisert av National Research Council. 1. Forsøksoppsett og Anestesi Utføre kalibreringen av de forskjellige signalene som skal fanges opp ved hjelp av et datainnsamlingssystem (trykk, temperatur, stempelforskyvning, elektro [EKG], kapnografi, etc.)….

Representative Results

Rottemodellen som beskrives her ble nylig anvendt for å sammenligne effektene av to inhibitorer av sarcolemmal natrium-hydrogen veksleren isoform 1 (NHE-1) på myokardial og hemodynamisk funksjon under brystkompresjoner og post-gjenoppliving 61. Det ble tidligere rapportert at NHE-1-hemmere dempe hjerteinfarkt reperfusjonsskade ved å begrense natrium-indusert cytosolisk og mitokondrie kalsium overbelastning, og dermed bidra til å bevare venstre ventrikkel utvidelsesevne under brystkompresjoner og dempe pos…

Discussion

Kritiske trinn i protokollen

Det er kritiske trinnene i protokollen. Når mestret, fortsette forberedelsene og protokoll som konsist beskrevet nedenfor. Den kirurgiske preparatet er meget flink, fremme katetre raskt gjennom små snitt utløser minimal eller ingen fartøy krampe og plassering av kateterspissen som forutsatt, fulgt av vellykket intubasjonsforhold etter en enkelt eller noen få forsøk (s); Således fullfører fremstillingen i ≈ 90 minutter fra den innledende dose pe…

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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