脊髓以上多巴胺D2受体介导左旋四氢巴马汀的镇痛作用

脊髓以上多巴胺D2受体介导左旋四氢巴马汀的镇痛作用 脊髓以上多巴胺D2受体介导左旋四氢巴马 汀的镇痛作用 ]SSNfY25~-9756ActaPh邮na.Sin中国药理学报]999Aug;20(8):715719 hUp?,.l’1~irnm*,*chiaainfogov~rL..’FrkMica[.715 SupraspinalD2receptorinvolvedinantin0cicepti0n inducedby1-tetrahydropalmatine HUJiangYuan,J1NGuo-Zhang~(DepartnwntPharmacolo~,,ShanghaiInstituteofMateriaMedica, ChineseAcademyofSciences,Shanghai200031,China) KEYWORDStewahydropalmatine;dopamine receptors;quinpirole;spiperone;SKF38393; Sch-23390;naloxoneianfinociception; non—narcoticanalgesics ABSTIT 删:Tostudythemleofdoparnine(DAjreceptors inl-tetrahydropalmatine(1-THP)一inducedantinocicep- tionM匝TH0DS:einwapedtonealLip)and intl”athecal”th)injectionswereusedtogivethedrugs. T}】etail-flicktestwasusedtoassessthenociceptive threshoIdofrats.REsu1:ByiPinjection. - THP(10,20,40mg’kg)aswellasD2receptor antagonistspiperone(1,2.3I’jproduced dose-dependentantinociceptiveeffectsonthenocicep- tionofrats,whilereceptoragonistquinpirole,Dt receptoragonistSKF38393.andDxreceptorantagonist Sch-23390showednoantinociceptionMoreover, l—Tl{orspiperone-inducedantinociceptionwas markedlyattenuatedbyquinpirole(2,3mg’kg..)but notSKF3gornaloxone.Ontheotherhand,ith quinpimle(20,30,40g’kg)alsoinducedadose- dependentantinociception.whileith?-THP. spiperone,SKF38393,andSch一23390didnotexhibit any.antinociceptionFurthermore,曲peronet20, 30,40g’)butnotSch-23390dose—dependently antugonisedtheanthiociceptioninducedbyquinpimle. 1-THP(,l00,200,300唱’jalsodranuatical- lyattenuatedthequinpirole—inducedantinociception withadose—dependentrelationship.C0NCLUS10N: ProjectsupportedbytheNationalNaturalScienceFoundationof China.3967t3~29and39600179.andStateKeyLaboratory ofDmgResearch.K016 CorrespondencetoProfJ1NGuo-Zhang Plm86216431一l端3.~xt,102Fax86-21—643. E-mailgzjin@se1%,ershcnc.acell Received1{)98-儿一10Accepted19994)’202 Activatingthespinalreceptororblockingthe supraspinalreceptorproducesantinociception. receptormightbenotdirectlyinvolvedinthe anthiociception.t-THP(asaantagonist)aswell asspiperoneproducesantinociceptionviablockingthe supraspinalreceptor. INTRODUCTION TeWahydropalmatine(THP)isthemainactive ingredientoftheCoo~talisambiguaChainetSch(or calledCoo,dalisturtshaninoviiBessfYanhuauo.YH ChouetCCHsu),afamousanalgesicoftraditional ChinesemedicineIts1evo-enatiomer{1-THP) possessestheanalgesicactionwithremarkablesedative manquilizingeffectL一1]}lPhasbeenactedasa remedyforanalgesicorsedation1istedintheChinese Pharmacopoeiaandtextbooksofphannacology. However.theexactanalgesicmechanismof1-THPstil1 remainsunclearuDtonow.Inthel)s.?一THPwas verifiedasadopamme(DA)receptorantagonistwith behavioral,biochemical,andelectrophysiologic experimentsl2,3J.andt-THPhadnoaffinityforoDiate receptors.Now.averyattractivetaskishowthe DAantagonisdceffectoff—THPelicitstheanalgesic actioninclinic.T0elucidutethismechanismnotonly wil1explaintheanalgesicmechanismof—THP.but alsowil1understandtheroleofDAnervoussystem involvedinanalgesia Muchevidencelmsshowedthatdopaminergic systemisdirectlyorindirectlyhivolvedintheprocesses ofnociceptionorantinociceptionIngeneral,most scholarsagreethattheprocessesofnociceptionatthe spinalorsupraspinallevelmightbeinfluencedbythe differentDAreceptorssubtypes’….However.many repomaboutD1andIreceptorareinvolvedin nociceptionwerediscordant.Somescholarsdemon— stratedthatD]andreceptorantagonistscould 716 IsSN雌_.J?56AcbPh棚ac0lSin Emaltaps@s日ersl~tLcaccn 中国药理学报1999Aug;20(8 Hm/Fax8,5,21_6474.2GX) enhancetheantinociceptiveeffectofrats,butother scholarsshowedthatmixedDl/D2orD2agonists producedadose—dependentantinocicepfiveeffect,while D.agonistSKF38393wasdevoidofsucheffect. RecentreportindicatedthatD2agonistquinpirole prolongednociceptivethresholdL….Therefoie,the mlethatDlandD2receptorsareinvolvedinnociception orantinocicepfion11~2dstobeclarifled.Fortiffsaim. thepresentstudyattemptedtocomparetheeffectsof 2一THP.selectiveDAreceptoragonistsandantagonists onnociceptionofratsbyintraperitoneal(ip)and inWathecal(ithjiniection.Inadd晡on,opioidergic receptorantagonistnaloxonewasalsousedtodetermine whe山erornotopioidtec?torsinfiuenced血e antinociceptiveeffectofl-THP. CH30 CH30 TdHlIlyd】?paI瑚dne MATERIAI.SANDMETHODS carriedout48hafteroperation. DrugsWereinjected slowlyviatllei山catlleteratavolumeof10., uL followedbvNS1OuLtoflushthecatheter Tai1nicktestThenociceptivethresholdwas assessedusingtail—flicktest.Thelalencyforaratto flickitstailawayfromasouw~ofradiantheatwas measuredwitllTailFlickTimer1.1(IITCInc,USA) thi’otl曲applyingnoxiousradiantheattostimulatethe blackenedundersurfaceofmiddletllirdportionoftlle taill—flicklatency(TFL)wasrecordedbvtlle di西”timer.Thebaselinelatency【BLjineachrat waskeptfrom3.0sto5.0s.ABLwasestablished bythreeWailsat5.minintervals.TheTFLofWailsat If)一minintervals协measuredafterinjectionofdrags. Eachgroupconsistedof6rats. StafisfiealanalysisTIwasconverted%of tllemaximalpossibleeffect.11%changeofTFL wascalculatedaccordingtotlleforl-llala.%C: (_,TFL—L)/TBL×100%.Data(x?s)were analyzedbvANOVAfollowedbvBonferronit-test. H3RESULTS MaterialsAdultSprague-Dawleyrats(t, 180—20Og)weresuppliedbyShanghaiAnimal Center,0daeseAcademyofSciences(Grade?, CertificateNo0O5)l-Tf口(mp141—142?,【JD 一 289.),isolatedbyShanghaiInstituteofMatefia Med.wasdissolved_nH2S0401mo1.L-..and adjustedtopH5.5with?OHO.1mol?L,. SKF38393,Sch一23390,quinpirole,spiperone,and naloxone,purchasedfromResearchBiochemicals IntemationalCompanyfUSA).wemdilutedwitll normalsaline(NS). 1ntrathecal(ith】injectionofanle,s catheterswereinstalledintothelumbarsubarachroid spaceforithinjectioneflv,ratswem anesthetizedwi山sodiumpentobarbital30mg?kg,a PE一10polyethylenecatheterof75IraTllongwas implantedintotllelranbarenlargementoftllespinal cord.Thoseratsshowinganymotordeficitresulting fromtllesurgicalprocedurewereexcludedfrom山e study.ExperimentswithithinjectionofdrugswGiv. Anfinecicepfiveeffectsofl-T日andn receptorantagonistspiperonebyipinjection AdmiifisWafionofipI-THP(10.20,40mg’kg) andD2receptorantagonistspiperone(1,2,3mg’ k2-.)producedsignificantanddose—dependent anfinociceptiveeffectsollthenociceptionofrats,while ipD2receptoragonistquinpirole(2.3Ingj,Dl receptoragonistSKFB8393(2,3mg’)and antagonistSch-23390【2,3mg’kg)showedno antinocicepfionAdministrationofiptllesamevolume ofvehicleorNSalsoexhibitediloeffectollnociception ofrats(T曲1,Fig1). D2receptoragonistquinpiroleattenuating theantlnocicepfioninducedbyipl-THPor spiperoneToevalttatethepossibleinteractionof 1-THP-induc酣antinociceptionwi山DAandopioidergic receptOrs,quinpirole,SKF38393,andnaloxonewere ipected5millafaripl-THP(40mg’kg..)or spiperone(3mg’kg).Quinpirole(2.3g’) producedadose—dependentantagonisticeffecton l-THE-orspipemne—inducedantinociception,while SKF38393【2,3mg’kg),naloxone【2,4mg? -.),andNSexhibitednoeffectolltheandnociceg- ISSN02?_956ActaPha力na?lSin E—mailaps@Ne/~,g.r.shcnccR 中国药理学报1999Aug;20(8l Phn/Fax8B0l一6474一蚴7】7 Tab1.EffectsofI-THP,DAreceptoragonistsand antagonistsontail?flicklalcy(TFL)ofm谯. DrugsDose%change ofTFL 1p DIagonistSKR393 DIantagonistSch-23390 a呈嘶qLdnpim|e antagonistspipemne NS 一 _rHP Vehicle 一 _rHP40mg’kg +quinpirole +NS +SKF38393 +na]OXOllle ith DtagonistSKR畦93 DIantagonistSch.23390 agonistquinpimle D2antagonistspipemne NS 一 _rHP Vehicle Qui~pim]e40JIg’ +st~pemne +NS +一_rHP +vehicle +Sch-23390 23mg’.. 2,3mg’.. 2.3mg’kg一. l,2,3mg’kg.. 0.4『nIl l0.?.40mg’kg 0.4mL 2,3mg’kg’ 04mL 2,3mg’kg一 2,4mg’kg’ ?,40旭’ ?.40g’ ?,3o,40g’ ?,40g’ l0L山 1O0,’200,300, ug’kg’ l0L山 ?.30,40JIg’ 10lfL 200,300;zg’kg. l0uL 20,40g’.. 十:denotesignificantincreaseofTFL:一:denote?changeof TFL;9:denolesigniticantantagonisticefl~t;+:denote? antagonisticeffect tioninducedbyf-n{Porspiperone(Tabl,Fig1). Antinociceptiveeffectsofithnreceptor agonlstquinpiroleatthespinallevel Administrationof|山agonistquinpirole(舶,3O,4o g’..)inducedasi~cantanddose-dependent antinociception,but山l-THP【100,200,3O0g’ kg).D2antagonistspiperone(20,40gkg),Dl agonistSKF38393【20,40g’kg),andDI antagonistSch-23390(20,40g’kg)showedno effectonnociception.ThesameadminisWalionof vehicleorNSalsoexhibitednoeffectonnociception (Tab1,Fig2). Dzreceptorantagonistspiperoneand 1.THPantagonizingithquinpirole-induced antinociceptionatthespinallevelToexplore whetherquinpirole-inducedantinociceptionismediated 鲁 8 Time,mill ng1.1~rcentagedqgesofTFLafteripl-THP 【A),bP<0.05.甲<0.01借vehicle;andipD2 receptoragomstquinpirolea恤nl】atedanllnoclcep? doninducedbipI-THPcB】.n=6.?s. Arrowdenotestimeatwhichipqulnplrole删NS. 中<0.05.rP<0.01珊I-THP. viaDxorrecel~orsatthespinallevel,Dlantagonist Sch?23390andantagonistspiperonewere|山injected 5minafterithquinpirole(40Pg.kg)respectively. Qninpirole—inducedantinociceptionwasdose-dependent— lyantagonisedbyspipemne(20,30,40g’kg)but notSch-23390(20,40gkg)orNSMoreover, 曲l-THP(200,300/*g’kg)alsoexhibited.皿 antagomsficeffectagainstquinpirole-inducedanti? nocicepfionwithadose—dependentrelationship,while itl1vehiclehadiioantagonisticeffect(Tabl,Fig2). DISCUSSION Inthepresentexperiments,thenociceptive thresholdofratswasmeosumdwithtai1-flicktest applyingnoxiousradiantheatstimuli.Theevidences ofl-THP,D1andagonistsorantagonistsithected directlyintospinallevelhavetx’esentedaconception thatatthespinalonlyagonistquinpirole,butRot 砷蚰蚰肿曲如钟0椰种”帅鲫神蛐如 7】8 ISSN2醋-977~PhmacSm E-m~l?s@??sI1acn 中画药理学报l9Ag;2OlB) Pt?Faxt36.21.6429 苫 目 巴 Time/min Fig2.Percentagedltng~iofTFLafterithl-THP (A);andithl-THPattenuatedantinociception inducedbyithreceptoragomstquinpirole(B). n=6.髯?s.Arrowdenotestimeatwhichith I-THPorvehicle.bp<O.05.<O.01quinpirole. - nlP,D2antagonist,Dtagonistandantagonist, participatesinantinocicepfion.Moreover,ourexperl’ mentstillshowedthatithantagonistspipeloneand f.11?butnotDlantagonistSch-23390COUldblockthe antinociceptioninducedbyagonistquinpimle TheresultsaIesimilartosomereportsindicatingthati山 DAagonistinhibiiedthespinalnociception,andi山 apomorphinecouldprolongthe1’FLofrats,.一】l’. Obviously,OUrfindingsstronglysupporttheideathat Dreceptorisinvolved-n山emodulationofnociception transmission,madthatactivatingthespinalD2receptor contributstotheantinociception.Thus,itispresumed thatanalgesicsiteofl-?口isriotatthespiI1allevel duetoits13,2antagonisticaction. Ontheotherhand,thepresentstudyresultssti11 showedthatipDe. antagonistspipemnecouldinducethe antinociception,whileipD2agorlistquinpirdiehadno antinociception.Tmsfactindicates山altheperipheral— lyadministrationofspipemneproducestheantinocicep— tionat山esupraspinallevelbutnotat山espinalleve1, jtsactionsitesofantinociceptiona佗1LIStoppositeto thoseofDzagonistqunipimleTheresuitsaresimilar tofilepreviousreport’orrecentreport,.More— over,ipspiperone-inducedantinuciceptionwasmarked- lyattenuatedbyipagonistquinpimlebutnotipDt agonistSch-T3390.Inotherwords,thepresentresults indicatethatreceptorinvolvedinantinociceptionjs dependent0ntheactionsites.bo血activatingthespinal D2receptorandblockingthesupraspinalD2receptor exerttheanalgesiceffect. Astothelx~sibleroleofD1receptorin nociceptionorantinociception,OlWexperimentshowed thatiporit11Dtagonistandantagonistdidnothaveany antinociceptionOurobservationisagreementwith otherreport-’.111eresultssuggestthatDtreceptor mightbenotdirectlyinvolvedintheantinociception I-THP,anovelDAreceptorantagonist,which actsasananalgesicdrug.alsoproducedantinociceptive effectbyipbutnotith.Interestingly,itht-THPcould alsodramaticallyattenuatetheantinociceptioninduced byithquinpirole.ThiseffectW~LSverysimilartoD2 antagonistspiperone.Itsuggeststhattheanalgesic effectofl-THPismediatedbyblockingthesupraspinal receptorInaddition,theresultthatipf-THP- Inducedantinociceptionisnotaaenuatedbynaloxone indicatesopioidergicreceptorsnotdirectlyinvolvedin theantmociceptionoff-THP. However.inthepresentstudy.itisdi街cultto asc~laintheexactactionsitesofl-THPinthebrain. Owingtothat?11?solubleintheacidicn~tnTlis difficulttobeadjustedtopH74forintmoerebmventri— cularoFIntro-nucleus~iection.wededu.ethatf-?{P— inducedantinociceptionismediatedbyblockingthe supraspInalD2receptorbasednotonlyonpresentstu results.butalsoontheotherresultsL.Inprevious experiment,usingFosproteinimmunohistochemistry technique.wehavedemonstratedthatl-THPactedon thesWiatum,accumbensnucleus,andsensofimotor cortext4J . Asweallknow,Fosprotein,aproductof immediateearlygenec-fosexpression,isamarkerof neuronalactivityfortracingneuronalactionsites Furthermore,f-THPshoweditsanalgesiceffectagainst formalin-painbasedonitsenhancingtheactb~ityof brainsterndescendingmodulationsystemTherefore. thesedatasupportthatfileanalgesicsitesoff-THPare atthesupraspinallevel Inconelusion.f-THPactsasaantagonist Involvedinantinuciceveeffectbyblocking山e 0{鲫曲钟栅仲0 ISSN0253-9756AbPharmacolSin中国药理学报1999】】g;20(8 E-rmilaps@set~,ershcncalPlm/Fax86-21~474—2629 supraspinalreceptorannmOi1painandacuptmCoareanalgesiainrats ActaPhysiolSin]998;50:49一. 13Fleetwood-WalkerSM,HopePJ,MitchellRAntinoci— REFERENCES.epdveactions0fdescendingaopmainergictractsoi1catand ratdorsalhornsnserBrleuroll~JPhysiol(Lrmd) 1KinKC,ZhellXF?HsuBStudiesoffthepharmacologica1]988 ;399:335—48 actionofcoryde0is?X?Theeffectsofopticalisomers14FluJY ,JinGZ.嗡Df?枷0pahna血earIaI[m oftewahydrop~lafine(THP)oncef血/l~lWOUSsystem?Fosexnindu?dbyfb?rlaliI卜pAc咀 ActaPhysiolSin1964;27:47—58PhaI_n ac0lSin]999;20:193—200 2XuSX—YuLP?HartYR,nYtJinGZE脓bof15M [不岛anJI.0删1TS山I附面[mc0u咖m 吼叫mopber越ondopanllI1eec印b母mtherv【?ssvstel/1:_盯v锄eI】toftheinduciblepro brain.ActaPhanmacolSin]989;10:104—10. ‘ 3HKX,JinGZ .. 啦0I1ic慨oftetrahydro- l;】4:421.5lpo 出 tol~er/nes..recep ; tom : :.芝一7,/e’toalstudiesSin1. 9913473843/出sciBl:一. 42~aang,JmGZ,XuSX,YuIJP,ChertY,Jia~gFY, a/.Effectsof1-stepholidineollcentralnervoll8and cardiovascularsystems. ActaPh~o]Sin19~6;7:522—6 5Fmssa-FjlhoR.RochaTB,ConceicaoIM,Mel1oCF, PereiraME.Effectsofdoparaine~icagentsonvisceml paintreasuredby虹writhingtest.ArchIntPlaarmaccdyn 1berl;331:74—93. 6Liuq6,OiJT.DafnyN.D2d0paln;T】ereceptor involvementinpinaldop~mne-inducedanfinociception Ijfesci1992;51:l485—92. 7一RoyJA,Standish,Tm’yLC.D0paeDl andreceptorantagonistspotentateanalgesicandmotet effectsofmolphinePh~olBiochemBehav1999;321 717—21 8.1eI]Se1]”IS.YakshTLEffectsofaIlintratheca1.agomst, apo~orplfine,oi1thermalandch.~tlcalevokednoxious response~inl’atsBrainRes1984;296:285—93. 9RoamDS.BoundsJK,AngcY.AdlooAA印i10le_ inducedaltemsoftailtemperattweappearashyperalgesia iIitheradiantheattail—flicktestPhamlacolBi~hernBehav 1998;59:77一艘. 10YakshTL.RudyTA.CI~oniccatheterizationofthespiilal subarachnoidspacePhysiol~ehav197,’6;17:1031—6. 1】BarasiS,DuggalKneeffectoflocalandsystec applicationof&vaminergicagentsontailflicklatencyin虹 rat.EurJPhamlaco1]985;JJ7:28T一94. 12GooX.XinBM.ZhucB.WuGc.XuSF.Eff~tof irara0~cali11je0nofdopo~~ecetnoragonists/ 脊髓以上多巴胺D2受体介导左旋四氢巴马汀的 镇痛作用 , (中国科学院上海药物研究所,上海2013031,中国) 关键词璺氢里里丕里堕至垡;喹吡罗; 螺哌隆;SKF38393;Sch.23390;纳洛酮; 抗痛效应;非麻醉性镇痛药 目的研究DA受体与左旋四氢巴马汀(1-THP)镇 痛作用的关系,以阐明l-THP的镇痛机制.方法: 腹腔(ip)与鞘内()给药,以大鼠甩尾反应观测热 伤害性致痛阐.结果:ipl-THP或受体拮抗剂 螺哌隆产生剂量依赖性镇痛效应,并能被受体 激动剂喹吡罗翻转,但不被纳洛酮翻转.而 l-THP或螺哌隆无镇痛效应,但它们能拮抗喹 吡罗引起的镇痛效?