Guide to Pharmacological Experiments全英班药理学实验教材.docx
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Guide to Pharmacological Experiments全英班药理学实验教材.docx
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GuidetoPharmacologicalExperiments全英班药理学实验教材
GuidetoPharmacologicalExperiments
DepartmentofPharmacology
GuangxiMedicalUniversity
2010.08.
Preface
PharmacologicalPracticalCourseisveryimportantforstudent,because
itnotonlyaffordsthembasicpharmacologicaltheory,butalsoletthemlearn
aboutsomebasictechniquesandmethodsofPharmacologyThispractical
courseincludesbothexperimentscarriedoutonisolatedtissuesororgansand
experimentsusingwholeanimals.Theseexperimentsaregoodguideto
students.
Wesincerelyhopethatthiscollectionofexperimentswillbeofrealvalue
toboththestaffofPharmocologyandstudents.Wewelcomeanycomments,
criticismsorsuggestionsfromourreaders.
DepartmentofPharmocology
Aug28,2010
CONTENTS
1.HandlingAnimalsandAdministration
2.One-compartmentopenmodel
3.Two-compartmentopenmodel
4.DeterminationofMedianEffectiveDose(ED50)ofSodiumPentobarbital
5.ComparingtheeffectsofPentobarbitalSodiumonMicebyDifferentRoutesofAdministration
6.DeterminationofHalf-lifeofDrugPlasmaConcentration
7.Dose-effectRelationshipofEffectofNAonBloodPressureinRabbit
8.IntoxicationandDetoxicationofOrganophosphorus
9.EffectsofDrugsAffectingtheSympatheticnervousSystemonBloodPressureinrabbits
10.Videorecordinganddiscussionontiledrugsofautonomicnervoussystemandβ–blockers
11.AnticonvulsantEffectoftheDrug
12.ObservationofAnalgesicActionofDrugswithTurningTrunkMethod
13.AntagonismofLidocaineontheArrhythmiaInducedbyOuabain
14.EffectofGlucocorticoidsontheInflammation
15.CasesDiscussion
ExperimentIHandlingAnimalsandAdministration
1.Rabbit
1)Holdingarabbit:
@Thecorrectholding:
Onehandgripsthescruffoftheneckfirmly,theother
handholdsthebuttockoftherabbit.
@Theerrorholdingincludes:
i.Onlygrasptheearsoftherabbitwithonehand.
ii.Onlygraspthebacklegsoftherabbitwithonehand.
2)Todistinguishanimalsaremaleorfemale:
forfemale,thedistancefrom
genitalporetoanusisshorterthanthatofthemale;youcanfinefivepairs
papillaeontheabdomen.Formale,thedistancefromgenitalporetoanusislongerthanthatofthefemale,insummeryoucanfindthetesteseasily.
3)Intravenousinjection(iv):
Inconsciousrabbits,injectionscanbemadeinto
themarginalveinoftheear.First,lookatthepictureoftherabbit’searon
TV.Thebluecolorsshowtheveinandtheredcolorshowstheartery.We
usuallyusetheveinoftheinnermarginoftheearasarouteof
administration.Therabbitisheldsuitablyorfixedinaboxandthefuris
removedfromtheveinatthetopoutsideedgeoftheeareitherbyplucking
orwithclippers.Itmaybenecessarytorubtheedgeoftheeargently.A
needlecaneasilybeinsertedintotheveinwhenitisdistendedandthe
plungerofthesyringepressedin.Asageneralprincipletheplungerofthe
syringeshouldbewithdrawntoseeifbloodappears.Iftheneedleisnot
correctlyplaceditisbesttoremoveitandtorepeattheattempt.Itisalways
wisesttostartasfarawayfromthebaseoftheearaspossibletoallowa
numberofattemptstobemade.Itisinhibitedthatthereisanyairinthevein
orneedle.Thespeedofpushingsyringeplungermustbeslow.
4)Asubcutaneousinjection(sc.):
Forasubcutaneousinjectiontheskinonthe
backoftherabbitisliftedup(likethatshowedontheTV)andtheneedle
pushedfirmlythroughit.Thepointoftheneedleshouldbequitefreeto
movebetweentheskinandthemuscle.Eithersideofthebackofananimal
ismostsuitableforasubcutaneousinjection.Ifthereisnooneavailabeto
holdtheanimal,itmaybenecessarytomaketheinjectionhigheruptowards
theheadbeauseonehandwillbeneededtoholdthescruftoftheneckof
docilerabbitswithoutassistance.Afterinjectionthereisalumponthesurface
theskin.
5)Anintramuscularinjection(im):
Foranintramuscularinjectionthe
procedureissimilartothesubcutaneousinjectionexceptthatthesitemustbe
amuscularpartofthebody,usuallyintheregionofthethighorbuttock.
Withthisinjectionthetipoftheneedleshouldnotbefreebutshouldhave
penetratedthemuscle.Afterinjectionthereisnotanylumponthesurfaceofistheskin.
6)Oralinjection:
WithrabbitsaNo.9rubbercatheterisused,insertedthrougha
holeinawoodenorplasticgagplacedbehindthefrontteeth.Thecatheteris
coveredwithglycerolandconnectedtoafunnelintowhichthedrugsolution
ispoured.Thecatheteristhencarefullyinsertedbetweenthetongueandthe
roofofthemouth.Oncepastthelarynxthecathetershouldmoveeasilyand
itmaybenecessarytomarkthetubetoindicatehowfaritshouldbeinserted.
Beforeadministrationyoushouldchecktheoperationisrightornotby
lookingatwhetherthesurfaceofthesolutionchanges(upordown)withthe
rabbit'srespiration.Ifnoanychangesthatmeansyoucaninjectthedrugto
therabbitbycatheter.Otherwise,thatmeansyouinsertthecatheterintothe
trachea.Thatisafaultyoperation.Faultyinjections(e.g.intothelungs)
usuallycauseimmediatedeath.
7)Dropdrugintotheeyes:
Whentherabbitisheldsuitably,onehandholdsa
lowereyelidoftherabbitasacup-shapewiththeforefingerandthumb.At
thesametime,thesamehandpressesthepunctalacrimaliawiththemiddle
finger.Anotherhanddropsthedrugintotheeye.Usually,thedrugisgiven
totheeyeabout1~---2drops.Afteradministrationkeepthedrugintheeye
foroneminute,thenloosetheeyelid.Noticethedrugsolutionisnotleaking
fromtheeye.
8)Intrathecalinjection:
Forintrathecalinjection,theneedleisinsertedintothe
vertebralcanalfromtheclearancebetweentheseventhlumbervertebraand
thefirsisacralbone.Thispositionislocatedonthemiddlepointoftheline
thatconnecttwosideiliaccrest,whentherabbit'sheadandhindlimbsare
curveddowntotheabdomen.Whentheneedleisinsertedintothevertebral
canal,therabbit'shindlimbsmakeajmnp.Thenthedrugisgiven.After
administration,thehindlimbsoftherabbitareparalysiswhiletheforelimbs
arenormal.
2.Rat
1)Holdingarat:
Ratsmaybeliftedbytheirtailsthoughtheyshouldbeputona
flatsurfaceassoonaspossible.Ifthisallowstheirfeetanygrip(whenthey
areputonaroughsurface),theywillthenpullawayfromwheretheyare
heldandthescruffoftheneckcanthenbegraspedintheotherhand.
2)Todistinguishanimalsaremaleorfemale:
forfemale,thedistancefrom
genitalporetoanusisshorterthanmale,youcanfindfivepairspapillaeon
abdomen.Formale,thedistancefromgenitalporetoanusislongerthan
femaleandtherearefursonthedistance,insummeryoucanfindthetestes.
3)Oralinjectioninarat:
Theratisheldwithonehandbygripingthescruffof
theneckwiththemouthheldsothattheesophagusisasstraightaspossible.
Theneedleisthencarefullyinsertedbetweenthetongueandtheroofofthe
mouth.Oncepastthelarynxtheneedleshouldmoveeasilyanditmaybe
necessarytomarktheneedletoindicatehowfaritshouldbeinserted
(usuallythisis3-4cm).Youmakesurethatoperationiscorrect,thenthe
drugisgiventotherat.Faultyinjections(e.g.intothelungs)usuallycause
immediatedeath.
4)Anintraperitonealinjection:
Foranintraperitonealinjectiontheratshouldbe
grippedbythescruffoftheneckandbyitstailorhindlimbs.Theneedleis
pushedfirmlythroughtheabdomeninaregionwhereitisnotlikelyto
penetratetheliver,kidney,spleenorbladderandtheplungerpressed.Then
thethumbpressestheplunger.
5)Determinetherat'sbodytemperature:
Thelefthandraisesthetailoftherat
andtheanusisexposed.Thethermometertopiscoveredwithvaseline
beforemeasure.Thethermometerisgentleinsertedintotheanusforlcm
depthandkeptthispositionwithrighthand.Afterremain3minutesinthe
rectumthethermometeristakenoutandthenumberisread.
6)Intravenousinjectionissimilartothatofthemouse.
3.Mouse
Itisthemostcommonusedanimalinthepharmacologicalexperiments.
1)Holdamouse:
Itisthesimilartothatofarat.Exceptthatmicecanevenbe
heldsingle-handedwiththeforefingerandthumbholdingthescruffofthe
neckandthelittlefingerholdingthetailorhindlegs.
2)Therelationshipofagesandthebodyweightofmice.Pleaselookatthe
tableontheTV.
3)Tonumbermouse:
Thesolutionofpicricacidisusedtonumberthemouse
onitsback.Thereareninesectionsonthebackofthemouse.Theright
forelimbofamouseisNo.I.Themiddleoftherightsideonthebackofa
mouseisNo.2.TherighthindlimbofamouseisNo.3.TheheadisNo.4.
ThemiddleofthebackinamouseisNo.5.TherootofthetailisNo.6.The
leftforelimbofamouseisNo.7.Themiddleoftheleftsideonthebackofa
mouseisNo.8.ThelefthindlimbofamouseisNo.9.No.10mousehasno
anypicricacidonthebodyanditisawholewhitemouse.(Pleaselookatthe
sketchmapontheTV).
4)Anintravenousinjection:
Inmice,intravenousinjectionismadeintothetail
vein.Firstlookattheanatomicalsketchmapofthemouse'stailatthe
horizontallevel(showedonTV).Thecartilageisinthemiddlepositionand
thearteryinthelow.Theveinsareontheaboveandtwosidesofthe
cartilage.Twosideveinsareoftenemployedtomakeintravenousinjection
becausetheyareclear.Themouseisplacedinatubethatisstopperedsothat
itstailprot
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