抗小鼠

Change in Hemoglobin Levels due to Anesthesia in Mice: An Important Confounder in Studies on Hematopoietic Drugs

Analgesicandanestheticdrugsmayhaveanimpactontheresultsachievedfromanimalexperiments.Inthestudypresentedhere,wetrytoenlightenwhetheranesthesiawithfentanyl/fluniasoneandmidazolam(HypnormandDormicum)hasaninfluenceonmeasurementsofhemoglobininmice.Inacross-overstudy,wehavecomparedhemoglobinlevelsintwogroupsofmice:anesthetizedversusnon-anesthetizedandfoundsignificantdecreaseinhemoglobinlevelsintheanesthetizedgroup(p < 0.05)unrelatedtowhichgroupreceivedtheanesthesia.ThemeanhemoglobinlevelsafterintraperitonealadmiNISTrationofHypnormandDormicumwas8.7mmol/Lcomparedtomeanhemoglobin9.9mmol/Lbeforeanesthesia(p < 0.001),andthedecreaselastedformorethan30min.Theseresultsshowthatanesthesiacanbeanimportantconfounderinstudiesinvolvingmeasurementsofhemoglobin,andthisshouldbetakenintoaccountwhenplanningstudiesandanalyzingdata.
KeyWords:anesthesia-hemoglobin-invivo-fentanyl-midazolam

Introduction

Whenperformingstudiesinvolvinglaboratoryanimals,appropriateanesthesiaiswarrantedalthoughcertaindrugsmayinfluenceontheresults.

Ithasbeenshownthatanesthetizedmicehaveadecreasedrespiratoryrateandmaydevelophypoxia,hypercapnia,andacidosis(1),allofwhichtheoreticallycanaffectthedatadependingofaimanddesignofthestudy.

Inourunit,weusethecombinationofHypnorm(fentanyl/fluniasone),whichisaneuroleptanalgesia,andasedativeDormicum(midazolam).ThiscombinationprovidessufficientdegreeofanesthesiaandanalgesiabuthassideeffectsasshownbyWhelanetal.(1).

Duringstudieswithgeneelectrotransfertomuscle(2,3)andskin(Gothelfetal,submittedforpublication)witherythropoietin(EPO)involvingsequentialbloodsamplingovertime,wehavediscoveredsomeinconsistencyregardingthebloodsamples.Attheendofthestudy,anunexpecteddecreaseinhemoglobinlevelscomparedtoearlierlevelswasobserved,theonlyobviousdifferencebeingthattheanimalswereanesthetized.

Wethusplannedtwostudiestoenlightenwhethertheanesthesiahasaninfluenceonthemeasurementofhemoglobin.


MaterialandMethods
Animals

FemaleNMRImice(ownbreed,CopenhagenUniversityHospitalHerlev),15weeksold,wereusedinthisstudy.Theanimalswerekeptinapathogen-freeenvironmentwith12h/12hlight/darknesscycleandfoodandwateradlibitum.ThestudywasperformedwithpermissionfromtheDanishAnimalExperimentsInspectorateandinaccordancewiththeEuropeanConventionfortheProtectionofVertebrateAnimalsusedforExperimentations.

Allmicewerehumanelyeuthanizedwithquickcervicaldislocationattheterminationofthestudy.

Anesthesia

Whenapplied,theanesthesiawasHypnorm(0.4ml/kg,JanssenSaunderton,Buckinghamshire,England)andDormicum(2mg/kg,Roche,Basel,Switzerland)dilutedinsterilewater.Theanesthesiawasadministeredasintraperitonealinjections.

MeasurementoftheHemoglobinLevels

HemoglobinwasanalyzedinmillimoleperliterfrombloodsamplesusingaHemoCueHb201+(HemoCue,AB,Sweden).Ahematocritstrawwasinsertedinthemedialangleoftheeye,directedalongthebonywallandtheplexusretro-orbitaliswaspunctured.AdropofbloodwasplacedonaHemoCueslideandanalyzedimmediately.

DesignofCross-OverStudy
ThedesignofthestudyisvisualizedinTable1.TwelvemiceweredividedintwogroupsAandB.Bloodsampleswerecollectedatday0andfromthenonceaweek.ThefirstfourbloodsampleswerecollectedduringanesthesiaingroupAandwithoutanesthesiaingroupB.Fromthefifthbloodsampleandtoterminationofthestudy,bloodsampleswerecollectedwithoutanesthesiaingroupAbutduringanesthesiaingroupB.

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