疼痛基礎研究方法

疼痛的動物模型與研究方法

AnimalModelsandMethods

InPainResearch萬有北京大學根底醫(yī)學院神經(jīng)生物學系12/29/2023NeuroscienceResearchInstitute,PekingUniversity1常用的動物模型神經(jīng)病理性痛模型神經(jīng)損傷：神經(jīng)瘤、慢性壓迫性損傷、局部神經(jīng)損傷、背根節(jié)慢性壓迫、低溫神經(jīng)損傷中樞神經(jīng)痛模型炎癥痛模型癌癥痛模型甩尾反射模型熱輻射或熱水甩尾機械刺激甩尾熱(冷)板反響模型內(nèi)臟痛模型化學誘導的軀體扭動模型膨脹結腸模型12/29/2023NeuroscienceResearchInstitute,PekingUniversity2常用的動物模型外周炎性痛模型皮膚炎性痛模型：Formalintest,BeeVenom致炎劑模型：白陶土-鹿角菜膠炎癥模型紫外線致炎扭體模型關節(jié)炎模型單關節(jié)炎模型多關節(jié)炎模型實驗型肌炎模型手術創(chuàng)傷模型12/29/2023NeuroscienceResearchInstitute,PekingUniversity3常用的動物模型炎癥痛模型外周炎性痛模型皮膚炎性痛模型：Formalintest,BeeVenom致炎劑模型：角叉菜膠模型紫外線致炎關節(jié)炎模型單關節(jié)炎模型多關節(jié)炎模型實驗型肌炎模型12/29/2023NeuroscienceResearchInstitute,PekingUniversity4常用的動物模型神經(jīng)病理性痛模型神經(jīng)損傷：神經(jīng)瘤、慢性壓迫性損傷、局部神經(jīng)損傷、背根節(jié)慢性壓迫、低溫神經(jīng)損傷中樞神經(jīng)痛模型內(nèi)臟痛模型化學誘導的軀體扭動模型膨脹結腸模型癌癥痛模型大鼠脛骨乳腺癌痛模型12/29/2023NeuroscienceResearchInstitute,PekingUniversity5AnimalmodelsofpainAcutestimulus-evokedpainThetail-flicktestThehot-platetestTheformalintestThepawflicktestImmersiontestforthermalhypersensitivityCold-allodyniatestThepin-pricktestformechano-hyperalgesiavonfreyHairtestformechano-allodyniaThewrithingtestTheDistensionofahollowviscusMusclepain12/29/2023NeuroscienceResearchInstitute,PekingUniversity6AnimalmodelsofpainModelsofchronicinflammatorypainAdjuvant-inducedarthritisUnilateralarthritisInflammationofahollowviscusUreteralcalculosis12/29/2023NeuroscienceResearchInstitute,PekingUniversity7扭體模型可采用小鼠或大鼠有多種刺激物都可誘發(fā)動物扭體(writhing)行為最常見的刺激物是醋酸(aceticacid)。將1克阿拉伯膠(arabicgum)參加9ml濃度為1%的醋酸溶液中，再注入實驗動物體內(nèi)，觀察注射后90分鐘期間每15分鐘內(nèi)出現(xiàn)典型扭體病癥的次數(shù)該模型可以模擬腹腔炎癥引起的腹痛病癥12/29/2023NeuroscienceResearchInstitute,PekingUniversity8TheAbdominalConstriction(Writhing)Test

tonicinflammatorypainspinallymediatedvisceral/subcutaneous0.9%AceticAcid(10ml/kg;intraperitoneal)12/29/2023NeuroscienceResearchInstitute,PekingUniversity9白陶土-鹿角菜膠炎癥模型白陶土(Kaolin)是一種細顆粒狀物質，成分為氧化鋁，起機械刺激作用；鹿角菜膠(carrageenan)是由水生植物鹿角菜中提取的膠體物質，具有過敏刺激作用。鹿角菜膠單獨實驗即可誘發(fā)炎癥，假設與白陶土合并使用，那么炎癥更為強烈可采用家兔或大鼠麻醉動物，由一側后肢足底注入4%白陶土混懸液0.1ml，并按摩5分鐘使之在組織中分散。在注射后1小時，再注入2%鹿角菜膠溶液0.05ml并按摩5分鐘。炎癥過程一般在第一次注射后2小時內(nèi)開始。動物后足紅腫，皮溫升高，PWT值降低等類似痛敏的病癥一般能持續(xù)12小時以上，24小時后根本復原。因而本模型屬于亞急性炎癥痛模型范圍本模型亦可采用關節(jié)腔注射12/29/2023NeuroscienceResearchInstitute,PekingUniversity10福爾馬林致痛模型模擬組織急性炎癥損傷所致的持續(xù)性疼痛大鼠或小鼠足底福爾馬林致痛模型：在動物一肢足底皮下注射稀釋的福爾馬林(formalin)溶液，動物的行為改變，如安靜時的屈腿、運動時的跛行以及舔足等。這些行為的程度(如舔足時間)與福爾馬林濃度成正比面部福爾馬林致痛模型：把不同濃度的福爾馬林溶液(0.2~10%)皮下注射到大鼠的右上唇，記錄注射后每3分鐘時間內(nèi)動物用同側前肢或后肢摩擦注射部位的秒數(shù)作為痛分數(shù)12/29/2023NeuroscienceResearchInstitute,PekingUniversity11福爾馬林致痛模型各種病癥普遍分為兩個時相：急性相或第一相：前5分鐘。之后有5-10分鐘的間歇持續(xù)相或第二相：15~60分鐘兩相均可用于實驗，但以第二相為常用。兩個時相的發(fā)生機制并不相同12/29/2023NeuroscienceResearchInstitute,PekingUniversity12慢性病理性疼痛慢性病理痛炎癥性痛(inflammatorypain)神經(jīng)病理性痛(neuropathicpain)癌癥痛(cancerpain)病理性痛時，共同存在：痛覺過敏(hyperalgesia):對傷害性刺激敏感性增強和反響閾值降低；觸誘發(fā)痛(allodynia):非痛刺激誘發(fā)持續(xù)性痛和自發(fā)痛(ongoingpainorspontaneouspain).12/29/2023NeuroscienceResearchInstitute,PekingUniversity13炎癥痛模型inflammatorypainmodel多發(fā)性佐劑關節(jié)炎模型含高濃度結核桿菌的福氏佐劑，向大鼠尾根部或足底作皮內(nèi)注射，一側或雙側后肢通常首先出現(xiàn)多個關節(jié)的炎癥單發(fā)性佐劑關節(jié)周圍炎模型完全福氏佐劑注射到動物后肢足底，造成單個關節(jié)周圍局部組織的炎癥反響單發(fā)性佐劑關節(jié)腔炎模型將高濃度的福氏佐劑直接注射到大鼠后肢踝關節(jié)腔中，引起一個具有急性、慢性兩相的高度局限的關節(jié)炎癥福氏佐劑關節(jié)炎模型福氏佐劑足底炎癥模型12/29/2023NeuroscienceResearchInstitute,PekingUniversity14Anklejoint:intra-articularinjectionofCFAWeek1:acuteperiodWeek2-3:subacuteperiodWeek4-9:chronicperiodChronicInflammatoryPainModel-Monoarthritis12/29/2023NeuroscienceResearchInstitute,PekingUniversity15012345690246810###//**********Scoresofextensionpaintest##############////////Time(weeksafterinjectionofCFA)IFA:IncompleteFreund'sAdjuvantCFA:CompleteFreund'sAdjuvantn=10/group*p<0.05,**p<0.01,***p<0.001comparedwithIFAgroup#p<0.05,##p<0.01,###p<0.001comparedwithleftankleChronicInflammatoryPainModel-MonoarthritisIFAleftCFAleftIFArightCFAright12/29/2023NeuroscienceResearchInstitute,PekingUniversity16AnimalmodelsofpainNeuropathicpainmodelsExperimentalanesthesiadolorosaExperimentalmodelsofpainfulperipheralneuropathyduetotraumatic,partialnervedamageChronicconstrictioninjuryPartialnervetransectioninjurySpinalnervetransectioninjuryExperimentalmodelsofpainfuldiabeticneuropathyChemotherapy-evokedpainfulperipheralneuropathy12/29/2023NeuroscienceResearchInstitute,PekingUniversity17NeuropathicpainfromnerveinflammationEliavandhiscolleagueshavedevelopedanenexperimentalmodelofaneuritis.TherataciaticnerveisexposedandlooselywrappedwithoxidizedcellulosethatissaturatedwithCFA.Within24and48htheanimalsdevelopheat-hyperalgesia,mechano-hyperalgesia,mechano-allodynia,and(toalesserdegree)cold-evokedpainslastuntil5to6daysaftertreatment,afterwhichresponsesallreturntonormal.(Eliav,E.etal.Neuropathicpainfromanexperimentalneuritisoftheratsciaticnerve.Pain1999;83:169)12/29/2023NeuroscienceResearchInstitute,PekingUniversity18L2L3L4L5L6L2L3L4L5Chung’sCCISeltzer12/29/2023NeuroscienceResearchInstitute,PekingUniversity1912/29/2023NeuroscienceResearchInstitute,PekingUniversity20Allodyniainratsinfectedwithvaricellazostervirus—asmallanimal

modelforpost-herpeticneuralgiaFollowingVZVinfectionoftheleftfootpadratsdevelopachronicmechanicalallodynia,whichispresentforlongerthan60dayspost-infectionandwhichresolvesby100dayspost-infection.Themodelisrobustandreproduciblewithanimalsconsistentlydevelopingallodyniaby3dayspost-infectionandcontinuingtopresentwithsymptomsforatleast30days.Thereproduciblenatureoftheinductionandcourseoftheallodyniaallowstheuseofthismodeltodeterminetheeffectofvariouscompoundson,andtoinvestigatethepathogenicmechanismsunderlyingthedevelopmentofVZV-inducedallodynia.ComparativestudiesusingHSV-1showthattheinductionofthechronicallodyniaisVZV-specificandisnotaresultisofvirusreplication-inducedtissuedamageoraccompanyinginflammation.12/29/2023NeuroscienceResearchInstitute,PekingUniversity21Fig.1.DurationofVZV-inducedallodynia12/29/2023NeuroscienceResearchInstitute,PekingUniversity22Fig.2.ReproducibilityofthemodelThemeanwithdrawalthresholdsobservedinfourindividualVZVstudies(n=24)arepresentedindividually(,,,

).Thedatafromthecontrols(n=24)fromthesefourstudieswerepooledandareplottedasasingleline(

).12/29/2023NeuroscienceResearchInstitute,PekingUniversity23Fig.3.SpecificityofthemodelAnimals(n=20)wereinfectedwith107pfuofHSV-1in50

lPBS.Controlanimals(n=6)receivedheat-inactivatedHSV-1.AllodyniawasassessedusinganelectronicvonFreyhairdailyuptoday6post-infection.Onegroup(n=10)ofinfectedanimalswastreatedwithvalaciclovir(50mg/kgtwicedailybyoralgavage)fromday0today6post-infection.Themeanwithdrawalthresholdsmeasuredingramsforweredeterminedipsilateralpawsandplottedagainsttimepost-infectionindaysforeachgroupandSEMshown.HSV-1(

),HSVplusvalaciclovir(

),control(

).(B)Animalswereinjectedinthelefthindpawonday0witheither4–8

106VZV-infectedCV-1cells(VZV,n=12)oruninfectedCV-1cells(control,n=6).Onegroup(n=6)ofinfectedanimalsweretreatedwithvalaciclovir(50mg/kgtwicedailybyoralgavage)fromday0today10post-infection.Themeanwithdrawalthresholdsmeasuredingramsweredeterminedforipsilateralpawsandplottedagainsttimepost-infectionindaysforeachgroupandSEMshown.VZV(

),VZVplusvalaciclovir(

),control(

).Thelineabovethegraphsindicatesthedurationofadministrationofvalaciclovir.12/29/2023NeuroscienceResearchInstitute,PekingUniversity24AnimalmodelsofpainVisceralpainmodelsColonic-rectaldistension(CRD)SmallboweldistensionArtificialkidneystonesUrinarybladderdistensionUrinarybladderirritantsIschemicstimuli(coronaryarteryocclusion)12/29/2023NeuroscienceResearchInstitute,PekingUniversity25Chemotherapy-evokedpainfulperipheralneuropathy(1)Painfulperipheralneuropathyisacommon,althoughseldomacknowledged,sideeffectofcancerchemotherapy.Chemotherapy-evokedneuropathicpainhasbeenmadeusingvincristineandpaclitaxel.Theuseofdosethatareconsiderablylowerthanthoseusedpreviously.Aleyetalinjectedvincristine5daysperweekfor2weeks.Theyfoundthatdosesof50and75g/kgproducedasignificantmechano-hyperalgesiabeginningaroundthetimeofthelastinjectiononday10andcontinuingforatleast12daysafterdosingceased.Bothdosesproducedasignificantlyincreasedthresholdtoheat-evokedpain.(AleyKO,etal.Vincristinehyperalgesiaintherat:amodelofpainfulcincristineneuropathyinhumans,Neuroscience1996;73:259)12/29/2023NeuroscienceResearchInstitute,PekingUniversity26Chemotherapy-evokedpainfulperipheralneuropathy(2)Polomanoetaldescribedapaclitaxel-evokedpainfulperipheralneuropathyintheratthatisnotassociatedwithanyevidenceofinjurytosensoryormotoraxonsandthatisnotaccompaniedbysignificanteffectsontheanimals’generalhealth.Ratsweretreatedwithpaclitaxelvia4i.p.injectionsgivenonalternatedayswithdosesof0.5,1.0,or2.0mg/kg.Allthreedosesproducedheat-hyperalgesia,mechano-hyperalgesia,mechano-allodynia,andcold-allodynia.Theabnormalpainsensationsbeganwithinseveraldaysoftheinitiationoftreatmentandlastedforatleastseveralweeksafterward.(PolomanoRC,etal.Apainfulperipheralneuropathyintheratproducedbythechemotherapeuticdrug,paclitaxel.Pain2001;94:293-304)12/29/2023NeuroscienceResearchInstitute,PekingUniversity27Colonic-RectalDistensionInrats,aflexiblelatexballoonfixedtoapliablecatheterispalcedintothedescendingcolonand/orrectumtransanally,securingthecathetertothetailwithtape.Briefly,eitheralatexcondomorafingerfromalatexglovemaybeusedastheballoon.ThecatheterinratsisTygon?flexibletubing.Fora7to8-cmlongballoon,6cmofoneendoftheflexibletubingisrepeatedlyperforatedwitha#35holepunch(20to25holes),insertedintheballoon,andtiedtightlywithsilksuture.(GebhartGF,etal.evaluationofvisceralpain,inMethodsinGastrointestinalpharmacology,Gaginella,TSEd,CRCPress,BocaRatom1996,359)12/29/2023NeuroscienceResearchInstitute,PekingUniversity28AnimalmodelsofpainModelsofcancerpain大鼠脛骨乳腺癌痛模型小鼠足底癌痛模型12/29/2023NeuroscienceResearchInstitute,PekingUniversity29癌痛實驗進展情況培養(yǎng)腫瘤細胞，建立癌癥痛模型行為學指標

痛覺過敏、痛覺超敏、自發(fā)性疼痛病理學指標

腫瘤形態(tài)大小、腫瘤病理切片、骨病理12/29/2023NeuroscienceResearchInstitute,PekingUniversity30小鼠腳掌皮膚癌痛模型動物：C57BL6,Male,6weeksoldB16-BL6(黑色素瘤細胞)模型組：右側腳掌皮下接種：B16-BL6105/20ul左側：0.1MPBS20ul對照組：右側:B16-BL6105/20ul〔heatkilled)左側:0.1MPBS20ulReference:SasamuraTetal.EurJPharmacol,200212/29/2023NeuroscienceResearchInstitute,PekingUniversity31小鼠腳掌腫瘤生長情況12/29/2023NeuroscienceResearchInstitute,PekingUniversity32疼痛的常見病癥人類的“疼痛〞與動物的“傷害性感受〞常見病癥：主要包括ongoingpainandstimulus-evokedpain自發(fā)痛(ongoingpain)誘發(fā)痛(stimulus-evokedpain)，包括痛覺過敏hyperalgesia和痛覺超敏〔觸誘發(fā)痛allodynia〕更為復雜的幻肢痛、鏡像痛、動物的自噬等動物模型上研究的策略是，通過觀察動物的行為，實驗者來推測動物是否發(fā)生了“疼痛〞12/29/2023NeuroscienceResearchInstitute,PekingUniversity33慢性痛的常見病癥自發(fā)痛spontaneouspain持續(xù)存在的通感覺痛覺過敏hyperalgesia弱的痛刺激引起強的痛感覺痛覺超敏allodynia，或稱觸誘發(fā)痛非痛刺激引起痛感覺12/29/2023NeuroscienceResearchInstitute,PekingUniversity34痛敏的種類與機制痛敏的種類(typesofhyperalgesia)痛敏包括痛覺過敏(hyperalgesia)與痛覺超敏(allodynia，也稱觸痛)原發(fā)性(primary)和繼發(fā)性(secondary)痛敏(hyperalgesia)繼發(fā)性痛敏：病區(qū)周圍非炎癥區(qū)也發(fā)生痛敏12/29/2023NeuroscienceResearchInstitute,PekingUniversity35⊙軸—軸反射末梢釋放SP+EAAPrimaryhyperalgesia原發(fā)性痛敏Secondaryhyperalgesia繼發(fā)性痛敏Allodynia痛覺超敏(觸痛)12/29/2023NeuroscienceResearchInstitute,PekingUniversity36PhilosophyofMeasuringPainThehumansubjectcanreporthissensationstous.Hedoessowithanact,somesortofbehavior-thespokenword,apencilmarkonaruledline,etc.Whatthenofmeasuringsensationinananimal?Theoptometrist’sprocedureisbasedontheimplicitassumptionthatmyprivatesubjectiveexperience(a“sharper〞image)isthesameaswhathewouldexperienceunderthesamecircumstances.12/29/2023NeuroscienceResearchInstitute,PekingUniversity37PhilosophyofMeasuringPainWeassumethatotherpeopleseelikeusbecausetheylooklikeus.Ratsdonotlooklikeus.Canwemaketheassumptionthatarat’sprivateandsubjectiveexperienceisIikeours?Initsbroadestsense,thequestionisdifficulttoansweranddependsonexactlywhatkindofexperiencewearediscussing.12/29/2023NeuroscienceResearchInstitute,PekingUniversity38PhilosophyofMeasuringPainWefindthattheaverageratheat-painthresholdisabout45°C.Itisalsotrueforahumanbeing.Thethresholdfordenaturationofmanyproteinsis45°CUndernormalcircumstances,thesensationofpainistightlyrelatedtotissuedamage.Itisreasonabletoarguethatthisrelationshiphasobviousevolutionaryvalue.Itisalsoanobviouslyprimitiverelationshipthatislikelytobehighlyconservedinman,rat,othermammals,andprobablyinallanimalswithanervoussystem.Thereispharmacologicalevidencethatarguesforthesimilaritybetweenpaininmanandothermammals:therankorderofthepotencyofopioidsisthesameasinhumanbeingsandrats.12/29/2023NeuroscienceResearchInstitute,PekingUniversity39MeasuringpaininanimalsAcuteandchronicpainThedistinctionisarbitrary“acute〞referstopainthatlastsforsecondstoaboutaday“chronic〞referstopainthatlastsforatleastseveraldays.Intheory,oncouldproduceanysortofinjurytoanybodypartintheanialanddeclarethatonehadapainmodelButpainfromdifferentcausesandfromdifferenttissuesmaybedissimilarinimportantways.Abdominalpainmaybeuniquelymodulatedbydrugsthatblockaopioid-likereceptors.12/29/2023NeuroscienceResearchInstitute,PekingUniversity40MethodsinPainResearchBehavioral:hot(cold)plate,vonFreyhair,painscorePharmacological:antagonist,radioligandbindingassayPsychologicalNeurochemical:neurotransmittercontentmeasurementwithhighperformanceliquidchromatography(HPLC)Cellular,molecular,andgeneticMorphological:Histochemical,immunohistochemical,fluorescentElectrophysiological:Extracellular,multi-channelrecordingpatchclampEvokedpotentialNon-invasive:PET,fMRICombinationofmethodsatdifferentlevels,integrationofabovemethods12/29/2023NeuroscienceResearchInstitute,PekingUniversity41單通道電流的記錄Neher&Sakmann(1976,1981)微玻管去神經(jīng)肌膜1991獲諾貝爾獎EricR.Kandel,etal.PrinciplesofNeuroscience4thedition.Fig.11-8.PatchClamp技術12/29/2023NeuroscienceResearchInstitute,PekingUniversity42影響傷害性感受測量的因素動物種類、品系、性別的選擇傷害性敏感度的晝夜變化身體不同部位的傷害性感受閾值得差異刺激區(qū)域的大小和連續(xù)刺激的間隔對閾值和反響的影響皮膚根底溫度對傷害性熱刺激閾值得影響12/29/2023NeuroscienceResearchInstitute,PekingUniversity43Strainandsexdifferencesinbasalthresholdinmice12/29/2023NeuroscienceResearchInstitute,PekingUniversity44測量疼痛的兩類方法第一類：測量產(chǎn)生傷害性反響所需的刺激的閾值。即設定一個標準反響，當發(fā)生了傷害性反響時，測定刺激的強度和時程。第二類：測量產(chǎn)生傷害性反響所需的刺激強度和時程。即刺激是標準化的。與第一類不同，它測定的不是閾值，而是反響的大小。12/29/2023NeuroscienceResearchInstitute,PekingUniversity45常用的痛刺激方法熱刺激冷刺激機械刺激化學刺激電刺激缺血12/29/2023NeuroscienceResearchInstitute,PekingUniversity46實驗動物的疼痛評價方法較理想的行為學評價方法應該具備能區(qū)分動物對傷害性和非傷害性刺激的不同反響痛刺激引起的行為反響隨刺激強度從痛閾到耐痛閾間出現(xiàn)相應改變測得的行為改變可以反映動物的痛感受動物的行為反響對鎮(zhèn)痛藥物的處理敏感能將非感覺性變化，如注意力、活動能力等與感覺性變化區(qū)分開反復刺激不引起或只引起極小的組織損傷12/29/2023NeuroscienceResearchInstitute,PekingUniversity47實驗動物的疼痛評價方法簡單的反射行為甩尾實驗(tailflicktest)鉀離子測痛法(Potassiumiontophoreticdolorimetry)缺血實驗：尾部束縛缺血后，動物搖頭、前肢回縮非訓練學會的組合行為熱板測痛法(hotplatetest)冷板測痛法(coldplatetest)翻滾實驗(wrotjomgresponse)發(fā)聲反響(colcalization)訓練學會的或自發(fā)反響逃跑或躲避反響(escapeandavoidancebehaviors)動機性選擇(motivationalchoiceparadign)12/29/2023NeuroscienceResearchInstitute,PekingUniversity48動物疼痛的行為學研究方法12/29/2023NeuroscienceResearchInstitute,PekingUniversity49外周神經(jīng)損傷后的機械痛敏OXOXOXCold-inducedOngoingPain

AfterPeripheralNerveInjury5

C12/29/2023NeuroscienceResearchInstitute,PekingUniversity51輻射熱甩尾測定痛閾〔電針〕12/29/2023NeuroscienceResearchInstitute,PekingUniversity5212/29/2023NeuroscienceResearchInstitute,PekingUniversity5312/29/2023NeuroscienceResearchInstitute,PekingUniversity5412/29/2023NeuroscienceResearchInstitute,PekingUniversity55012345690246810###//**********Scoresofextensionpaintest##############////////Time(weeksafterinjectionofCFA)IFA:IncompleteFreund'sAdjuvantCFA:CompleteFreund'sAdjuvantn=10/group*p<0.05,**p<0.01,***p<0.001comparedwithIFAgroup#p<0.05,##p<0.01,###p<0.001comparedwithleftankleChronicInflammatoryPainModel-MonoarthritisIFAleftCFAleftIFArightCFAright12/29/2023NeuroscienceResearchInstitute,PekingUniversity5612/29/2023NeuroscienceResearchInstitute,PekingUniversity5712/29/2023NeuroscienceResearchInstitute,PekingUniversity5812/29/2023NeuroscienceResearchInstitute,PekingUniversity5912/29/2023NeuroscienceResearchInstitute,PekingUniversity6012/29/2023NeuroscienceResearchInstitute,PekingUniversity61TheAbdominalConstriction(Writhing)Test

tonicinflammatorypainspinallymediatedvisceral/subcutaneous0.9%AceticAcid(10ml/kg;intraperitoneal)12/29/2023NeuroscienceResearchInstitute,PekingUniversity6212/29/2023NeuroscienceResearchInstitute,PekingUniversity6312/29/2023NeuroscienceResearchInstitute,PekingUniversity64疼痛的研究方法疼痛研究是現(xiàn)代神經(jīng)科學研究的一局部從傳統(tǒng)的行為學、藥理學、臨床觀察，到電生理學、神經(jīng)化學，以及到現(xiàn)代的細胞學、組織學、分子生物學、影像學、蛋白質組的方法臨床研究遵循隨機、對照、多中心，以及志愿、雙盲等根本原那么根底研究的多學科方法行為學藥理學〔包括腦內(nèi)核團立體定位注射、蛛網(wǎng)膜下腔注射等〕生理學〔包括電生理學〕細胞學解剖學〔如神經(jīng)示蹤〕與組織學〔包括一般組織學與免疫組織化學〕生物化學和分子生物學基因組學和蛋白質組學……12/29/2023NeuroscienceResearchInstitute,PekingUniversity65脊髓丘腦束神經(jīng)元中樞敏化痛敏，通覺超敏〔allodynia)遞質釋放配體/受體反響跨膜信號轉導基因轉錄因子基因激活SPEAANK1RNMDARNO/CGMPNO/PKGPKCPKAMARPKCREBFOS通道、受體基因表達痛敏傷害性刺激短時程中時程長時程12/29/2023NeuroscienceResearchInstitute,PekingUniversity67Vanilloidreceptortype1(VR1)Distribution

inDRGinNormalRatsIB4VR1mergedScalebar=40μm

12/29/2023NeuroscienceResearchInstitute,PekingUniversity68ChangeofVR1ExpressioninDRGafterCFAInjectionAverageVR1-ir=averagegrayvalue(meandensity)-backgroundn=3,*p<0.05**p<0.01background12/29/2023NeuroscienceResearchInstitute,PekingUniversity69CorrelationbetweenHyperalgesiaandVR1ProteinLevel

12/29/2023NeuroscienceResearchInstitute,PekingUniversity70PAIN12/29/2023NeuroscienceResearchInstitute,PekingUniversity71Threedifferenttypicalpatternsofectopicdischarges12/29/2023NeuroscienceResearchInstitute,PekingUniversity72ChangesofectopicdischargesafterSNL12/29/2023NeuroscienceResearchInstitute,PekingUniversity73Correlationanalysisbetweentactileallodyniaandectopicactivityinthefirst24hoursafterSNL12/29/2023NeuroscienceResearchInstitute,PekingUniversity74Correlationanalysisbetweentactileallodyniaandectopicactivityinthedays1-14afterSNL12/29/2023NeuroscienceResearchInstitute,PekingUniversity75Neuropathicpain:

EarlyspontaneousafferentactivityisthetriggerAfterCCInerveinjury,bothA-fibers(c)andC-fibers(f)displayedhighlevelsofspontaneousactivityintheCCI-ratsthatwereuntreated(‘na?¨ve’)orperfusedwithsaline,buttreatmentwitheitherbupivacaineorTTXduringthefirst4–7dayspost-injuryinhibitedthiselevationinspontaneousactivity.

WenruiXie,JudithA.Strong,JohannaT.A.Meij,Jun-MingZhang,LeiYu.Painxx(2005)1–1412/29/2023NeuroscienceResearchInstitute,PekingUniversity76AllodyniaOpioids--C.N.S.CentralSensitization+NMDA(Pain)(Touch=Pain)NE5-HT12/29/2023NeuroscienceResearchInstitute,PekingUniversity77Wind-upofaWDRneuron12/29/2023NeuroscienceResearchInstitute,PekingUniversity78InductionofLTPofC-fiber-evokedpotentials

in