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.
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.
Tæt på 360.000 personer i USA 1 og mange flere i hele verden 2 lider en episode med pludseligt hjertestop hvert år. Forsøg på at genoprette livet kræver ikke kun, at hjerteaktivitet blive genetableret, men at skader på vitale organer forhindres, minimeres, eller omvendt. Nuværende genoplivningsudstyr teknikker giver en indledende genoplivning på ca. 30%; dog overlevelse til udskrivelse fra hospitalet er kun 5% 1. Myokardiedysfunktion, neurologisk dysfunktion, systemisk inflammation, tilstødende sygdomme, eller en kombination deraf forekommende efter genoplivning konto for stor andel af patienter, der dør på trods af indledende afkast på cirkulation. Således er større forståelse for de underliggende patofysiologi samt nye genoplivning tilgange presserende behov for at øge hastigheden af initial genoplivning og efterfølgende overlevelse med intakt organfunktion.
Animal tilstandls af hjertestop spiller en kritisk rolle i udviklingen af nye genoplivning behandlingsformer ved at give indsigt i patofysiologien af hjertestop og genoplivning og tilbyde praktiske midler at konceptualisere og afprøve nye tiltag, før de kan blive testet på mennesker 3. Rotten model af lukkede brystet genoplivningsudstyr (CPR) beskrevet her har spillet en vigtig rolle. Modellen er udviklet i 1988 af Irene von Planta – en forskningsstipendiat på det tidspunkt – og hendes samarbejdspartnere 4 i laboratoriet af afdøde Professor Max Harry Weil MD, Ph.D. ved University of Health Sciences (omdøbt Rosalind Franklin University of Medicine and Science i 2004) og har været flittigt brugt i forbindelse med genoplivning overvejende af stipendiater professor Weil og deres praktikanter.
Modellen simulerer en episode med pludseligt hjertestop med genoplivning forsøgt ved traditionelle CPR teknikker og omfatter således indføringion af ventrikelflimmer (VF) ved at levere en elektrisk strøm til højre ventrikel endocardium og levering af lukkede brystet CPR med en pneumatisk drevet stempel-enheden, mens samtidig leverer positivt tryk ventilation med ilt-beriget gas. Ophør af VF opnås ved transtorakal levering af elektrisk stød. Rotten model skaber en balance mellem modeller udviklet i store dyr (fx svin) og modeller udviklet i mindre dyr (fx mus), som tillader udforskning af nye forskningsresultater begreber i et godt standardiseret, reproducerbar og effektiv måde med adgang til en robust oversigt over relevante målinger. Modellen er særlig nyttig i tidlige stadier af forskning for at udforske nye koncepter og undersøge virkningerne af konfoundere før gennemførelse af undersøgelser hos større dyremodeller, som er dyrere, men større translationel virkning.
En Medline søgning på alle peer-reviewed artikler rapporteringimilar rottemodel har VF som mekanismen for hjertestop og en form for lukket bryst genoplivning afslørede i alt 69 yderligere oprindelige undersøgelser under anvendelse af modellen, da den først blev offentliggjort i 1988 4. De forskningsområder omfatter patofysiologiske aspekter af genoplivning 5-17, faktorer, der påvirker resultater 18-30, rolle farmakologiske interventioner undersøger karkontraherende midler 31-43, buffer agenter 44, inotropika 45, agenter sigte på myocardial eller cerebral beskyttelse 46-70, og også virkningerne af mesenchymstamceller 71-73.
Modellen og protokol beskrevet i denne artikel anvendes i øjeblikket på Resuscitation Institute. Men der er flere muligheder for at "tilpasse" den model baseret på den kapacitet til rådighed for de enkelte efterforskere og målene for undersøgelserne.
Kritiske trin i protokollen
Der er kritiske trin i protokollen. Når mestrer, fortsæt forberedelse og protokol som kortfattet beskrevet nedenfor. Den kirurgiske forberedelse er hurtig, fremrykkende katetre hurtigt gennem små indsnit udløser minimal eller ingen fartøj spasmer og positionering kateterspidserne efter hensigten, efterfulgt af en vellykket trakealintubation efter en enkelt eller nogle få forsøg (s); således, færdiggøre forberedelserne i ≈ 90 min fra den indled…
The authors have nothing to disclose.
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.
Name of Material/ Equipment | Company | Catalog Number | Comments/Description |
Sodium pentobarbital | Sigma Aldrich | P3761 | http://www.sigmaaldrich.com/catalog/product/sigma/p3761?lang=en®ion=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 |