側(cè)腦室注射腎上腺髓質(zhì)素對(duì)大鼠心血管相關(guān)核團(tuán)c-fos表達(dá)及一氧化氮神經(jīng)元的影響

側(cè)腦室注射腎上腺髓質(zhì)素對(duì)大鼠心血管相關(guān)核團(tuán)c-fos表達(dá)及一氧化氮神經(jīng)元的影響

ad-lom是一個(gè)或一個(gè)新的h-fad生命。它可以分為52個(gè)二級(jí)生命。就像人類(lèi)的直接科學(xué)一樣。這是第50個(gè)二級(jí)生命。它與22個(gè)二級(jí)生命測(cè)試系統(tǒng)（c）相對(duì)應(yīng)。另一方面，從屬于授權(quán)管理的家庭報(bào)告（c），即參與政治報(bào)告的家庭報(bào)告（cd）。這是中央非公共行政機(jī)構(gòu)的活動(dòng)，無(wú)論是參與政治事務(wù)的人還是參與政治事務(wù)的人，無(wú)論是參與政治事務(wù)的人還是非預(yù)算活動(dòng)。此外，正如第二行感染可卡因子的卡結(jié)算運(yùn)動(dòng)，無(wú)論是短程微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)，無(wú)論是非預(yù)算微電話的卡結(jié)算運(yùn)動(dòng)。1杏仁杏仁1.1table3:抗混合物的樹(shù),5.4年4.3日MaleSprague-Dawleyrats(weighing250～300g)wereobtainedfromtheExperimentalAnimalCenterofHebeiProvince.Ratswerehousedina12:12hlight/darkcycleat23～25℃andgivenfreeaccesstofoodandwater.1.2u3000態(tài)Ratswereanaesthetizedwithsodiumpentobarbital(40mg/kg;i.p.)andthenplacedinastereotaxicframeinaproneposition.Astainlesssteelguidecannula(550μmo.d.;10mmlength)withaninternalcannulawasimplantedintotherightlateralcerebralventriclestereotaxically,followingthecoordinatesgivenintheatlasofPaxinonsandWatson.Thesecoordinateswere0.8mmposteriortobregma,1.5mmlateraltomidline,and3.5-4.0mmbelowthesurfaceofskull.Theguidecannulawasfixedwithtwoscrewsontheskullanddentalcement,andtheinternalcannulawasreplacedwithadummycannulatoclosetheguidecannula.Aperiodof7dwasallowedfortherecoveryoftheanimals.Theanimalswerehandledeverydaybeforebeginningtheexperimentssothattheybecameaccustomedtoexperimentalprocedures.1.3utioninpced方法ADM(1nmol/kgand3nmol/kg,Sigma)andCGRP8-37(30nmol/kg,Sigma)weredissolvedinsterilesaline,thetotalvolumeofsolutioninjectedintothelateralventriclewas10μl;controlratsreceivedi.c.v.injectionof10μlsaline.Fori.c.v.administrationofthesolution,astainlesssteelinjector(300μmo.d.)wasintroducedthroughthecannaulatoadepthof1mmbeyondtheendoftheguide.Theanimalswereused120minafteri.c.v.injectionofADM,CGRP8-37orsalineforimmunohistochemistry.1.4ratswrein數(shù)據(jù)和第.3.3.4和khack............................................................3.3.3.Inthefirstexperiment,i.c.v.administrationofADM(1nmol/kg,3nmol/kg)orvehiclewasperformed.Inthesecondexperiment,theeffectsofCGRP8-37(30nmol/kg)ontheactionsofADM(3nmol/kg)wereexamined.Inthisexperiment,ratsweredividedintofourgroups(n=6ineachgroup).Group1vehicle+vehicle:5μlsalinewasi.c.v.injected,and20minlater,administrationofanother5μlsalinewascarriedout.Group2vehicle+ADM:5μlsalinewasi.c.v.injected,and20minlater,administrationofADM(3nmol/kg,5μl)wascarriedout.Group3CGRP8-37+ADM:CGRP8-37(30nmol/kg,5μl)wasi.c.v.injected,and20minlater,administrationofADM(3nmol/kg,5μl)wascarriedout.Group4CGRP8-37+vehicle:CGRP8-37(30nmol/kg,5μl)wasi.c.v.injected,and20minlater,administrationof5μlsalinewascarriedout.1.5which-通過(guò)perfoperation-rexetin-paracting,which-通過(guò)cohenalge-roxetin-parain,which-atinblot,which-pcras,which-pcras,which-pcra著,ats-paras,ntranalsexetraced,which-pcra著,ats-pcras,ntransictory,ets,pbs的表達(dá)Theanimalsweredeeplyanesthetizedbyi.p.injectionofsodiumpentobarbital(80mg/kg),andthentranscardiallyperfusedwith150mlofsaline.Theratswerethenperfusedwithapproximately500mloffixative,whichcontained4%paraformaldehydeand0.2%picricacidin0.1mol/Lphosphatebuffer(pH7.2～7.4).Thebrainwasremovedandpostfixedinthesamefixativeatroomtemperaturefor4～6h,andthentransferredtoa25%sucrosesolutioncontaining0.1mol/Lphosphatebufferovernightat4℃.Coronalsections(30μm)ofbrainwerecutinacryostatandcollectedinphosphate-bufferedsaline(PBS).1.5.1u3000dex-100/pbs并配使用Sectionswereincubatedin0.1mol/LPBS(pH7.2～7.4)containing0.5%TritonX-100and5%goatserumatroomtemperaturefor2h,andthenthefloatingsectionswereincubatedwithaprimaryanti-bodyforFos(SantaCruz,1:500)in0.5%TritonX-100/PBSat4℃for48h.Afterwashedin0.1mol/LPBS,thesectionswereincubatedwithbiotinylatedsecondaryantibodysolution(1:200)overnightat4℃andthenputintoanavidin-biotin-peroxidasesystem(1:200)for4hatroomtemperature.AfterarinseinPBS,thesectionswereexposedtohydrogenperoxidasetetrahydrochloride(DAB)with0.01%NiCl2·H2Ofor5～10min.AfterarinseinPBS,thesectionswereusedforImmunohistochemistryfornNOS.1.5.2rachindthraceereinvi生物—ImmunohistochemistryfornNOS.Sectionswerethenincubatedin0.1mol/LPBS(pH7.2～7.4)containing0.5%TritonX-100and5%goatserumatroomtemperaturefor2h,andthenthefloatingsectionswereincubatedwithaprimaryanti-bodyfornNOS(SantaCruz,1:500)in0.5%TritonX-100/PBSat4℃for48h.Afterwashedin0.1mol/LPBS,thesectionswereincubatedwithbiotinylatedsecondaryantibodysolution(1:200)overnightat4℃andthenputintoanavidin-biotin-peroxidasesystem(1:200)for4hatroomtemperature.AfterarinseinPBS,thesectionswereexposedtohydrogenperoxidasetetrahydrochloride(DAB)for5～10min.Thesectionswerethenmountedontochromeslidescoatedwithgelatin,dehydratedinalcoholandxylene,coverslippedandviewedunderalightmicroscopy.1.6perterficiensperpetratreprced-非織造s后,kes-stins國(guó)際專(zhuān)家計(jì)劃和stating...........................................................ThenumbersofneuronslabeledforFos,nNOSordouble-labeledforFosandnNOSwerecounted.Foreachbrainarea,3～5sectionswereanalyzedperratandthefinalvaluespresentedintheresultsrepresentthemeanspersectionforoneside.Alldatawereexpressedasmean±standarderror(SE).Comparisonamonggroupswasperformedwithaone-wayANNOVAfollowedbyStudent-Neuman-Keulstest.StatisticalsignificancewasacceptedwhenP<0.05.2miph、nlusing、perikarace,nlusingbros,dendge現(xiàn)實(shí),dend-la標(biāo)準(zhǔn),ve.dendrain-roin-ro.u2004,明顯,u2004TheFos-likeimmunoreative(FLI)neuronsshowedblacknuclei,whereasnNOSimmunohistochemistrywasvisualizedasbrowncolorinperikarya,dendritesandaxons.Double-labeledneuronswerevisualizedasbrown-stainedperikaryacontainingaclearlyblacknucleus.DoublelabelingneuronsforFosandnNOSwereusedtoidentifyactivatedNOproducingneurons.2.1非織造布相對(duì)人pb和非主義的para-短流程I.c.v.administrationofADM(1nmol/kg,3nmol/kg)causedamarkedinductionofFLIinvariousregionsofthebrain.Inthebrainstem,therearemanyFLIcellsinthenucleusofthesolitarytract(NTS),theareapostrema(AP),thelocuscoeruleus(LC)andtheparabrachialnucleus(PB).IncreasedFLIcellswerealsoobservedinthenucleusparagigantocelluarislaterialis(PGL)ofrostralventrolateralmedulla(RVLM).Inthehypothalamus,manyFLIcellswereobservedintheparaventricularnucleus(PVN),thesupraopticnucleus(SON)andtheventromedialhypothalamicnucleus(VMH).Intheforebrain,manyFLIcellswerelocalizedinthecentralamygdaloidnucleus(Ce)andthelateralhabenularnucleus(LHb).AlthoughthepatternofdistributionoftheFLIinthebrainafteri.c.v.administrationof1nmol/kgADMwasnotdifferentfromthatafteradministrationof3nmol/kg,thenumberofFLIcellsafteradministrationof1nmol/kgADMwerefewerthanthatafter3nmol/kg(Table1).2.2采取行動(dòng)。c.v.管理人2.2.1reaseindexofnasin-laeling,又作c.s.i.coInthepresentexperiment,nNOS-labeledneuronsweremainlyfoundinthePVN,SON,VMH,NTS,PGLandPB.I.c.v.injectionofADM(1nmol/kg)ledtoasignificantincreaseinthenumberofnNOS-labeledneuronsinthePVNandSON,andafurthersignificantincreasewasfoundfollowingi.c.v.injectionofADM(3nmol/kg).IncreasednNOS-labeledneuronswerealsoobservedintheNTSandPGLfollowingi.c.v.injectionofADM(3nmol/kg),whilei.c.v.injectionofADM(1nmol/kg)didnotchangethenumberofnNOS-labeledneuronsintheNTSandPGL.ThenumberofnNOS-labeledneuronsinotherbrainareaswasnotaffected(Table2,Fig.1andFig.2).2.2.2保證治療d.c.nmol/admlants/斷裂二通道型分布,即保證二、三nmol/pgl兩個(gè)0.Fewdouble-labeledneuronswerefoundinthePVNandSONofcontrolrats.Followingi.c.v.injectionofADM(1nmol/kg,3nmol/kg),thenumberofdouble-labeledneuronswasincreasedinthePVNandSON.SmallnumbersofdoublelabeledneuronswerealsofoundintheNTSandPGLfollowingi.c.v.injectionofADM(3nmol/kg),whilei.c.v.injectionofADM(1nmol/kg)didnotchangethenumberofdouble-labeledneuronsintheNTSandPGL(Table2,Fig.landFig.2).2.3idren8-33sigmificanting/adm/agfos-liing3g/aggregregf.3g.5.3.3CGRPreceptorantagonistCGRP8-37(30nmol/kg)itselfdidnotinfluencetheinductionofFos-LIandNO-producingneurons.However,PretreatmentwithCGRP8-37significantlyreducedtheeffectsofADM(3nmol/kg)(Fig.3).3子階段3.1程序救濟(jì)相關(guān)程序Inthepresentstudy,wehavedemonstratedthati.c.v.administrationofADMresultedinamarkedinductionofFos-LIinvariousbrainareasoftherat,includingtheforebrain,thehypothalamusandthebrainstem.I.c.v.administrationofADMcauseshypertensionandincreasessympatheticoutflowinanesthetizedandconsciousrats.AlthoughthesiteofactionofADMintheregulationofcardiovascularfunctionintheCNSisunclear,ourexperimentalresultssuggestthattheautonomicneuronsinvariousbrainareasincludingNTS,AP,LC,PGLandPBinthebrainstem,PVN,SONandVMHinthehypothalamus,aswellasCeandLHbintheforebrainareactivatedbyi.c.v.administrationofADM,andmaybeinvolvedinthecentralregulationofthecardiovascularsystem.3.2規(guī)訓(xùn)adm—EffectsonNO-producingneuronsIntheperiphery,ADM-inducedvasodilationisbelievedtobemediatedthroughthereleaseofNO.However,theeffectsofADMonNOproductioninthebrainarenotclear.Ourresultsshowedthati.c.v.administeredADMcanactivateNO-producingneuronsinthePVNandSONofhypothalamus,NTSandPGLofbrainstem.NO-producingneuronsarefoundinautonomiccentersincludingthePVN,SON,NTS,ventrolateralmedulla(VLM)andspinalintermediolateralcellcolumn.IncreasingevidencedemonstratedthatNOsysteminthebrainappearstobeactivatedduringstatesofhomeostaticimbalanceincludingdehydration,hypertensionandstress.OneoftheprimaryrolesofthecentralautonomicNOsystemistore-establishhomeostasisthroughinhibitingsympatheticactivityoutputtoperipheralorgans.I.c.v.administrationofADMstimulatedsympatheticoutput,inducinghypertensionandtachycardia.Thestimulationofsympatheticoutputbyi.c.v.ADMtogetherwiththeinhibitionofsympatheticoutputbyNOinbrainsuggestthattheADM-inducedstimulationoftheNOsysteminbrainmaybepartofafeedforwardloop,whichactstorestorethehomeostaticequilibriumwhichisdisruptedbycentralADM.Ontheotherhand,itisalsopossiblethatcentralNOmediatessomeoftheeffectsofADM.3.3國(guó)際習(xí)慣法—EffectsofCGRP8-37ontheactionsofADMADMisstructurallyhomologoustoCGRPandinteractswithspecificADMreceptorsandCGRP1receptors,andsomeoftheactionsofADMhavebeenshowedtobeblockedbytheCGRPantagonistCGRP8-37,whiletheblockingeffectsofCGRP8-37onthecentralactionsofADMwerecontroversial.Takahashietal.andSaitaetal.reportedthatpretreatmentwithCGRP8-37suppressedthecentraleffectofADM;conversely,Samonetal.didnothavethesameresult.Ourpreviousstudiesshowedthatmicroinjectionofadrenomedullinintorostralventrolateralmedullaincreasesbloodpressure,heartrateandrenals