疼痛基礎(chǔ)研究方法

1、萬有北京大學(xué)基礎(chǔ)醫(yī)學(xué)院神經(jīng)生物學(xué)系1/16/2022Neuroscience Research Institute, Peking University1常用的動(dòng)物模型常用的動(dòng)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、部分神經(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(jī)械刺激甩尾熱(冷)板反應(yīng)模型內(nèi)臟痛模型化學(xué)誘導(dǎo)的軀體扭動(dòng)模型膨脹結(jié)腸模型1/16/2022Neuroscience Research Institute, Peking University2常用的動(dòng)物模型常用的動(dòng)物模型外周炎性痛模型皮膚炎性痛模型：Formal

2、in test, Bee Venom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(guān)節(jié)炎模型單關(guān)節(jié)炎模型多關(guān)節(jié)炎模型實(shí)驗(yàn)型肌炎模型手術(shù)創(chuàng)傷模型1/16/2022Neuroscience Research Institute, Peking University3常用的動(dòng)物模型常用的動(dòng)物模型炎癥痛模型外周炎性痛模型皮膚炎性痛模型：Formalin test, Bee Venom致炎劑模型：角叉菜膠模型紫外線致炎關(guān)節(jié)炎模型單關(guān)節(jié)炎模型多關(guān)節(jié)炎模型實(shí)驗(yàn)型肌炎模型1/16/2022Neuroscience Research Institute, Peking University4常用的動(dòng)物模

3、型常用的動(dòng)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、部分神經(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(xué)誘導(dǎo)的軀體扭動(dòng)模型膨脹結(jié)腸模型癌癥痛模型大鼠脛骨乳腺癌痛模型1/16/2022Neuroscience Research Institute, Peking University5Animal models of painAcute stimulus-evoked painThe tail-flick testThe hot-plate testThe formalin testThe paw flick testImmersion test for ther

4、mal hypersensitivityCold-allodynia testThe pin-prick test for mechano-hyperalgesiavon frey Hair test for mechano-allodyniaThe writhing testThe Distension of a hollow viscusMuscle pain1/16/2022Neuroscience Research Institute, Peking University6Animal models of painModels of chronic inflammatory painA

5、djuvant-induced arthritisUnilateral arthritisInflammation of a hollow viscusUreteral calculosis1/16/2022Neuroscience Research Institute, Peking University7扭體模型扭體模型可采用小鼠或大鼠有多種刺激物都可誘發(fā)動(dòng)物扭體(writhing)行為最常見的刺激物是醋酸(acetic acid)。將1克阿拉伯膠(arabic gum)加入9ml濃度為1%的醋酸溶液中，再注入實(shí)驗(yàn)動(dòng)物體內(nèi)，觀察注射后90分鐘期間每15分鐘內(nèi)出現(xiàn)典型扭體癥狀的次數(shù)該模型可以

6、模擬腹腔炎癥引起的腹痛癥狀1/16/2022Neuroscience Research Institute, Peking University8The Abdominal Constriction (Writhing) Test tonic inflammatory pain spinally mediated visceral/subcutaneous0.9% Acetic Acid(10 ml/kg; intraperitoneal)1/16/2022Neuroscience Research Institute, Peking University9白陶土白陶土-鹿角菜膠炎癥模型鹿角菜

7、膠炎癥模型白陶土(Kaolin)是一種細(xì)顆粒狀物質(zhì)，成分為氧化鋁，起機(jī)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(zhì)，具有過敏刺激作用。鹿角菜膠單獨(dú)實(shí)驗(yàn)即可誘發(fā)炎癥，若與白陶土合并使用，則炎癥更為強(qiáng)烈可采用家兔或大鼠麻醉動(dòng)物，由一側(cè)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(shí)，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(shí)內(nèi)開始。動(dòng)物后足紅腫，皮溫升高，PWT值降低等類似痛敏的癥狀一般能持續(xù)12小時(shí)以上，24小時(shí)后基本復(fù)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(guān)節(jié)腔注射1/1

8、6/2022Neuroscience Research Institute, Peking University10福爾馬林致痛模型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(dòng)物一肢足底皮下注射稀釋的福爾馬林(formalin)溶液，動(dòng)物的行為改變，如安靜時(shí)的屈腿、運(yùn)動(dòng)時(shí)的跛行以及舔足等。這些行為的程度(如舔足時(shí)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(0.210%)皮下注射到大鼠的右上唇，記錄注射后每3分鐘時(shí)間內(nèi)動(dòng)物用同側(cè)前肢或后肢摩擦注射部位的秒數(shù)作為痛分?jǐn)?shù) 1/16/2022Neuroscience Researc

9、h Institute, Peking University11福爾馬林致痛模型福爾馬林致痛模型各種癥狀普遍分為兩個(gè)時(shí)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：1560分鐘兩相均可用于實(shí)驗(yàn)，但以第二相為常用。兩個(gè)時(shí)相的發(fā)生機(jī)制并不相同1/16/2022Neuroscience Research Institute, Peking University12慢性病理性疼痛慢性病理性疼痛慢性病理痛炎癥性痛(inflammatory pain)神經(jīng)病理性痛(neuropathic pain)癌癥痛(cancer pain)病理性痛時(shí)，共同存在：痛覺過敏(hyperalges

10、ia): 對傷害性刺激敏感性增強(qiáng)和反應(yīng)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(ongoing pain or spontaneous pain).1/16/2022Neuroscience Research Institute, Peking University13炎癥痛模型炎癥痛模型 inflammatory pain model多發(fā)性佐劑關(guān)節(jié)炎模型含高濃度結(jié)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(cè)或雙側(cè)后肢通常首先出現(xiàn)多個(gè)關(guān)節(jié)的炎癥 單發(fā)性佐劑關(guān)節(jié)周圍炎模型完全福氏佐劑注射到動(dòng)物后肢足底，造成單個(gè)關(guān)節(jié)周圍局部組織的炎癥反應(yīng)單發(fā)性佐劑關(guān)節(jié)腔炎模

11、型將高濃度的福氏佐劑直接注射到大鼠后肢踝關(guān)節(jié)腔中，引起一個(gè)具有急性、慢性兩相的高度局限的關(guān)節(jié)炎癥 福氏佐劑關(guān)節(jié)炎模型福氏佐劑足底炎癥模型1/16/2022Neuroscience Research Institute, Peking University14Ankle joint:intra-articular injection of CFAWeek 1: acute periodWeek 2-3: subacute periodWeek 4-9: chronic periodChronic Inflammatory Pain Model - Monoarthritis1/16/2022Ne

12、uroscience Research Institute, Peking University15012345690246810#/*Scores of extension pain test#/Time (weeks after injection of CFA)IFA: Incomplete Freunds AdjuvantCFA: Complete Freunds Adjuvantn=10 / group*p0.05, *p0.01, *p0.001 compared with IFA group#p0.05, #p0.01, #p0.001 compared with left an

13、kleChronic Inflammatory Pain Model-MonoarthritisIFA leftCFA left IFA rightCFA right1/16/2022Neuroscience Research Institute, Peking University16Animal models of painNeuropathic pain modelsExperimental anesthesia dolorosaExperimental models of painful peripheral neuropathy due to traumatic, partial n

14、erve damageChronic constriction injuryPartial nerve transection injurySpinal nerve transection injuryExperimental models of painful diabetic neuropathyChemotherapy-evoked painful peripheral neuropathy1/16/2022Neuroscience Research Institute, Peking University17Neuropathic pain from nerve inflammatio

15、nEliav and his colleagues have developed an en experimental model of a neuritis. The rat aciatic nerve is exposed and loosely wrapped with oxidized cellulose that is saturated with CFA. Within 24 and 48 h the animals develop heat-hyperalgesia, mechano-hyperalgesia, mechano-allodynia, and (to a lesse

16、r degree) cold-evoked pains last until 5 to 6 days after treatment, after which responses all return to normal. (Eliav, E. et al. Neuropathic pain from an experimental neuritis of the rat sciatic nerve. Pain 1999; 83:169)1/16/2022Neuroscience Research Institute, Peking University18L2L3L4L5L6L2L3L4L5

17、ChungsCCISeltzer1/16/2022Neuroscience Research Institute, Peking University191/16/2022Neuroscience Research Institute, Peking University20Allodynia in rats infected with varicella zoster virusa small animalmodel for post-herpetic neuralgiaFollowing VZV infection of the left footpad rats develop a ch

18、ronic mechanical allodynia, which is present for longer than 60 days post-infection and which resolves by 100 days post-infection. The model is robust and reproducible with animals consistently developing allodynia by 3 days post-infection and continuing to present with symptoms for at least 30 days

19、. The reproducible nature of the induction and course of the allodynia allows the use of this model to determine the effect of various compounds on, and to investigate the pathogenic mechanisms underlying the development of VZV-induced allodynia. Comparative studies using HSV-1 show that the inducti

20、on of the chronic allodynia is VZV-specific and is not a result is of virus replication-induced tissue damage or accompanying inflammation.1/16/2022Neuroscience Research Institute, Peking University21Fig. 1. Duration of VZV-induced allodynia1/16/2022Neuroscience Research Institute, Peking University

21、22Fig. 2. Reproducibility of the modelThe mean withdrawal thresholds observed in four individual VZV studies (n=24) are presented individually (, ,). The data from the controls (n=24) from these four studieswere pooled and are plotted as a single line ().1/16/2022Neuroscience Research Institute, Pek

22、ing University23Fig. 3. Specificity of the modelAnimals (n=20) were infected with 107 pfu of HSV-1 in 50 l PBS. Control animals (n=6) received heat-inactivated HSV-1. Allodynia was assessed using an electronic von Frey hair daily up to day 6 post-infection. One group (n=10) of infected animals was t

23、reated with valaciclovir (50 mg/kg twice daily by oral gavage) from day 0 to day 6 post-infection. The mean withdrawal thresholds measured in grams for were determined ipsilateral paws and plotted against time post-infection in days for each group and SEM shown. HSV-1 (), HSV plus valaciclovir (), c

24、ontrol (). (B) Animals were injected in the left hindpaw on day 0 with either 48106 VZV-infected CV-1 cells (VZV, n=12) or uninfected CV-1 cells (control, n=6). One group (n=6) of infected animals were treated with valaciclovir (50 mg/kg twice daily by oral gavage) from day 0 to day 10 post-infectio

25、n. The mean withdrawal thresholds measured in grams were determined for ipsilateral paws and plotted against time post-infection in days for each group and SEM shown. VZV (), VZV plus valaciclovir (), control (). The line above the graphs indicates the duration of administration of valaciclovir.1/16

26、/2022Neuroscience Research Institute, Peking University24Animal models of painVisceral pain modelsColonic-rectal distension (CRD)Small bowel distensionArtificial kidney stonesUrinary bladder distensionUrinary bladder irritantsIschemic stimuli (coronary artery occlusion)1/16/2022Neuroscience Research

27、 Institute, Peking University25Chemotherapy-evoked painful peripheral neuropathy (1)Painful peripheral neuropathy is a common, although seldom acknowledged, side effect of cancer chemotherapy.Chemotherapy-evoked neuropathic pain has been made using vincristine and paclitaxel. The use of dose that ar

28、e considerably lower than those used previously. Aley et al injected vincristine 5 days per week for 2 weeks. They found that doses of 50 and 75 g/kg produced a significant mechano-hyperalgesia beginning around the time of the last injection on day 10 and continuing for at least 12 days after dosing

29、 ceased. Both doses produced a significantly increased threshold to heat-evoked pain. (Aley KO, et al. Vincristine hyperalgesia in the rat: a model of painful cincristine neuropathy in humans, Neuroscience 1996; 73: 259)1/16/2022Neuroscience Research Institute, Peking University26Chemotherapy-evoked

30、 painful peripheral neuropathy (2)Polomano et al described a paclitaxel-evoked painful peripheral neuropathy in the rat that is not associated with any evidence of injury to sensory or motor axons and that is not accompanied by significant effects on the animals general health. Rats were treated wit

31、h paclitaxel via 4 i.p. injections given on alternate days with doses of 0.5, 1.0, or 2.0 mg/kg. All three doses produced heat-hyperalgesia, mechano-hyperalgesia, mechano-allodynia, and cold-allodynia. The abnormal pain sensations began within several days of the initiation of treatment and lasted f

32、or at least several weeks afterward. (Polomano RC, et al. A painful peripheral neuropathy in the rat produced by the chemotherapeutic drug, paclitaxel. Pain 2001; 94: 293-304)1/16/2022Neuroscience Research Institute, Peking University27Colonic-Rectal DistensionIn rats, a flexible latex balloon fixed

33、 to a pliable catheter is palced into the descending colon and/or rectum transanally, securing the catheter to the tail with tape. Briefly, either a latex condom or a finger from a latex glove may be used as the balloon. The catheter in rats is Tygon flexible tubing . For a 7 to 8-cm long balloon, 6

34、 cm of one end of the flexible tubing is repeatedly perforated with a #35 hole punch (20 to 25 holes), inserted in the balloon, and tied tightly with silk suture. (Gebhart GF, et al. evaluation of visceral pain, in Methods in Gastrointestinal pharmacology, Gaginella, TS Ed, CRC Press, Boca Ratom 199

35、6, 359)1/16/2022Neuroscience Research Institute, Peking University28Animal models of painModels of cancer pain大鼠脛骨乳腺癌痛模型小鼠足底癌痛模型1/16/2022Neuroscience Research Institute, Peking University29癌痛實(shí)驗(yàn)進(jìn)展情況癌痛實(shí)驗(yàn)進(jìn)展情況 培養(yǎng)腫瘤細(xì)胞，建立癌癥痛模型培養(yǎng)腫瘤細(xì)胞，建立癌癥痛模型?行為學(xué)指標(biāo)行為學(xué)指標(biāo) 痛覺過敏、痛覺超敏、自發(fā)性疼痛?病理學(xué)指標(biāo)病理學(xué)指標(biāo) 腫瘤形態(tài)大小、腫瘤病理切片、骨病理1/16/2022N

36、euroscience Research Institute, Peking University30小鼠腳掌皮膚癌痛模型小鼠腳掌皮膚癌痛模型動(dòng)物：C57BL6, Male, 6 weeks oldB16-BL6 (黑色素瘤細(xì)胞)模型組：右側(cè)腳掌皮下接種：B16-BL6 105/20ul 左側(cè)： 0.1M PBS 20ul對照組：右側(cè): B16-BL6 105/20ul（heat killed) 左側(cè): 0.1M PBS 20ulReference: Sasamura T et al. Eur J Pharmacol, 20021/16/2022Neuroscience Research In

37、stitute, Peking University31小鼠腳掌腫瘤生長情況小鼠腳掌腫瘤生長情況1/16/2022Neuroscience Research Institute, Peking University32疼痛的常見癥狀疼痛的常見癥狀人類的“疼痛”與動(dòng)物的“傷害性感受”常見癥狀：主要包括ongoing pain and stimulus-evoked pain自發(fā)痛(ongoing pain )誘發(fā)痛(stimulus-evoked pain)，包括痛覺過敏hyperalgesia和痛覺超敏（觸誘發(fā)痛allodynia）更為復(fù)雜的幻肢痛、鏡像痛、動(dòng)物的自噬等動(dòng)物模型上研究的策略是，

38、通過觀察動(dòng)物的行為，實(shí)驗(yàn)者來推測動(dòng)物是否發(fā)生了“疼痛”1/16/2022Neuroscience Research Institute, Peking University33慢性痛的常見癥狀慢性痛的常見癥狀自發(fā)痛spontaneous pain持續(xù)存在的通感覺痛覺過敏hyperalgesia弱的痛刺激引起強(qiáng)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1/16/2022Neuroscience Research Institute, Peking University34痛敏的種類與機(jī)制痛敏的種類與機(jī)制痛敏的種類(types of hyperalgesia)痛敏包括痛覺過

39、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1/16/2022Neuroscience Research Institute, Peking University35軸軸反射末梢釋放 SP+EAAPrimary hyperalgesia原發(fā)性痛敏Secondary hyperalgesia繼發(fā)性痛敏Allodynia痛覺超敏 (觸痛)1/16/2022Neuroscience Research Institute, Peking Univers

40、ity36Philosophy of Measuring PainThe human subject can report his sensations to us. He does so with an act, some sort of behavior- the spoken word, a pencil mark on a ruled line, etc.What then of measuring sensation in an animal? The optometrists procedure is based on the implicit assumption that my

41、 private subjective experience (a “sharper” image) is the same as what he would experience under the same circumstances. 1/16/2022Neuroscience Research Institute, Peking University37Philosophy of Measuring PainWe assume that other people see like us because they look like us. Rats do not look like u

42、s. Can we make the assumption that a rats private and subjective experience is Iike ours? In its broadest sense, the question is difficult to answer and depends on exactly what kind of experience we are discussing.1/16/2022Neuroscience Research Institute, Peking University38Philosophy of Measuring P

43、ainWe find that the average rat heat-pain threshold is about 45C. It is also true for a human being.The threshold for denaturation of many proteins is 45CUnder normal circumstances, the sensation of pain is tightly related to tissue damage. It is reasonable to argue that this relationship has obviou

44、s evolutionary value. It is also an obviously primitive relationship that is likely to be highly conserved in man, rat, other mammals, and probably in all animals with a nervous system.There is pharmacological evidence that argues for the similarity between pain in man and other mammals: the rank or

45、der of the potency of opioids is the same as in human beings and rats.1/16/2022Neuroscience Research Institute, Peking University39Measuring pain in animalsAcute and chronic painThe distinction is arbitrary“acute” refers to pain that lasts for seconds to about a day“chronic” refers to pain that last

46、s for at least several days.In theory, on could produce any sort of injury to any body part in the anial and declare that one had a pain modelBut pain from different causes and from different tissues may be dissimilar in important ways.Abdominal pain may be uniquely modulated by drugs that block a o

47、pioid-like receptors.1/16/2022Neuroscience Research Institute, Peking University40Methods in Pain ResearchBehavioral: hot (cold) plate, von Frey hair, pain scorePharmacological: antagonist, radio ligand binding assayPsychologicalNeurochemical: neurotransmitter content measurement with high performan

48、ce liquid chromatography (HPLC)Cellular, molecular, and geneticMorphological: Histochemical, immunohistochemical, fluorescentElectrophysiological:Extracellular, multi-channel recordingpatch clampEvoked potentialNon-invasive: PET, fMRICombination of methods at different levels, integration of above m

49、ethods1/16/2022Neuroscience Research Institute, Peking University41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(jiǎng)Eric R. Kandel, et al. Principles of Neuroscience4th edition. Fig. 11-8.Patch Clamp 技術(shù)1/16/2022Neuroscience Research Institute, Peking University42影響傷害性感受測量的因素影響傷害性感受測量的因素

50、動(dòng)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應(yīng)的影響皮膚基礎(chǔ)溫度對傷害性熱刺激閾值得影響1/16/2022Neuroscience Research Institute, Peking University43Differences of basal thresholds amonginbred strains of mice129AAKR B6B10 B/c C3H C58 RIIIS SM23456n=46-80StrainsBasal threshold (s)Sex difference of basal t

51、hresholdsamong inbred strains129AAKRB6B10B/cC3HC58RIIISSM23456n=14-51StrainsBasal threshold (s)Strain and sex differences in basal threshold in mice1/16/2022Neuroscience Research Institute, Peking University44測量疼痛的兩類方法測量疼痛的兩類方法第一類：測量產(chǎn)生傷害性反應(yīng)所需的刺激的閾值。即設(shè)定一個(gè)標(biāo)準(zhǔn)反應(yīng)，當(dāng)發(fā)生了傷害性反應(yīng)時(shí)，測定刺激的強(qiáng)度和時(shí)程。第二類：測量產(chǎn)生傷害性反應(yīng)所需的刺激強(qiáng)

52、度和時(shí)程。即刺激是標(biāo)準(zhǔn)化的。與第一類不同，它測定的不是閾值，而是反應(yīng)的大小。1/16/2022Neuroscience Research Institute, Peking University45常用的痛刺激方法常用的痛刺激方法熱刺激冷刺激機(jī)械刺激化學(xué)刺激電刺激缺血1/16/2022Neuroscience Research Institute, Peking University46實(shí)驗(yàn)動(dòng)物的疼痛評價(jià)方法實(shí)驗(yàn)動(dòng)物的疼痛評價(jià)方法較理想的行為學(xué)評價(jià)方法應(yīng)該具備能區(qū)分動(dòng)物對傷害性和非傷害性刺激的不同反應(yīng)痛刺激引起的行為反應(yīng)隨刺激強(qiáng)度從痛閾到耐痛閾間出現(xiàn)相應(yīng)改變測得的行為改變可以反映動(dòng)物的痛感受動(dòng)

53、物的行為反應(yīng)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(dòng)能力等與感覺性變化區(qū)分開反復(fù)刺激不引起或只引起極小的組織損傷1/16/2022Neuroscience Research Institute, Peking University47實(shí)驗(yàn)動(dòng)物的疼痛評價(jià)方法實(shí)驗(yàn)動(dòng)物的疼痛評價(jià)方法簡單的反射行為甩尾實(shí)驗(yàn)(tail flick test)鉀離子測痛法(Potassium iontophoretic dolorimetry)缺血實(shí)驗(yàn)：尾部束縛缺血后，動(dòng)物搖頭、前肢回縮非訓(xùn)練學(xué)會(huì)的組合行為熱板測痛法(hot plate test)冷板測痛法(cold plate test)翻滾實(shí)驗(yàn)(wrot

54、jomg response)發(fā)聲反應(yīng)(colcalization)訓(xùn)練學(xué)會(huì)的或自發(fā)反應(yīng)逃跑或躲避反應(yīng)(escape and avoidance behaviors)動(dòng)機(jī)性選擇(motivational choice paradign)1/16/2022Neuroscience Research Institute, Peking University48動(dòng)物疼痛的行為學(xué)研究方法動(dòng)物疼痛的行為學(xué)研究方法1/16/2022Neuroscience Research Institute, Peking University49外周神經(jīng)損傷后的機(jī)械痛敏OXOXOXCold-induced Ongoin

55、g Pain After Peripheral Nerve Injury5 C1/16/2022Neuroscience Research Institute, Peking University51輻射熱甩尾測定痛閾（電針）輻射熱甩尾測定痛閾（電針）1/16/2022Neuroscience Research Institute, Peking University521/16/2022Neuroscience Research Institute, Peking University531/16/2022Neuroscience Research Institute, Peking Uni

56、versity541/16/2022Neuroscience Research Institute, Peking University55012345690246810#/*Scores of extension pain test#/Time (weeks after injection of CFA)IFA: Incomplete Freunds AdjuvantCFA: Complete Freunds Adjuvantn=10 / group*p0.05, *p0.01, *p0.001 compared with IFA group#p0.05, #p0.01, #p0.001 c

57、ompared with left ankleChronic Inflammatory Pain Model-MonoarthritisIFA leftCFA left IFA rightCFA right1/16/2022Neuroscience Research Institute, Peking University561/16/2022Neuroscience Research Institute, Peking University571/16/2022Neuroscience Research Institute, Peking University581/16/2022Neuro

58、science Research Institute, Peking University591/16/2022Neuroscience Research Institute, Peking University601/16/2022Neuroscience Research Institute, Peking University61The Abdominal Constriction (Writhing) Test tonic inflammatory pain spinally mediated visceral/subcutaneous0.9% Acetic Acid(10 ml/kg

59、; intraperitoneal)1/16/2022Neuroscience Research Institute, Peking University621/16/2022Neuroscience Research Institute, Peking University631/16/2022Neuroscience Research Institute, Peking University64疼痛的研究方法疼痛的研究方法疼痛研究是現(xiàn)代神經(jīng)科學(xué)研究的一部分從傳統(tǒng)的行為學(xué)、藥理學(xué)、臨床觀察，到電生理學(xué)、神經(jīng)化學(xué)，以及到現(xiàn)代的細(xì)胞學(xué)、組織學(xué)、分子生物學(xué)、影像學(xué)、蛋白質(zhì)組的方法臨床研究遵循隨機(jī)、

60、對照、多中心，以及志愿、雙盲等基本原則基礎(chǔ)研究的多學(xué)科方法行為學(xué)藥理學(xué)（包括腦內(nèi)核團(tuán)立體定位注射、蛛網(wǎng)膜下腔注射等）生理學(xué)（包括電生理學(xué)）細(xì)胞學(xué)解剖學(xué)（如神經(jīng)示蹤）與組織學(xué)（包括一般組織學(xué)與免疫組織化學(xué)）生物化學(xué)和分子生物學(xué)基因組學(xué)和蛋白質(zhì)組學(xué)1/16/2022Neuroscience Research Institute, Peking University65脊髓丘腦束神經(jīng)元中樞敏化脊髓丘腦束神經(jīng)元中樞敏化痛敏，通覺超敏痛敏，通覺超敏 （allodynia)SPEAANK1RNMDARNO/CGMPNO/PKGPKCPKAMARPKCREBFOS通道、受體基因表達(dá)痛敏傷害性刺激1/16/2022N