p3疼痛基礎(chǔ)的研究法

1、11/9/2021neuroscience research institute, peking university1疼痛的動(dòng)物模型與研究方法animal models and methodsin pain research 萬(wàn)有北京大學(xué)基礎(chǔ)醫(yī)學(xué)院神經(jīng)生物學(xué)系11/9/2021neuroscience research institute, peking university2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

2、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11/9/2021neuroscience research institute, peking university3常用的動(dòng)物模型 外周炎性痛模型 皮膚炎性痛模型：formalin test, bee venom 致炎劑模型：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11/9/2021neuroscience research institute, peking university4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,

3、 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11/9/2021neuroscience research institute, peking university5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11/9/2021neuroscience research institute, peking university6animal models of p

4、ain acute stimulus-evoked pain the tail-flick test the hot-plate test the formalin test the paw flick test immersion test for thermal hypersensitivity cold-allodynia test the pin-prick test for mechano-hyperalgesia von frey hair test for mechano-allodynia the writhing test the distension of a hollow

5、 viscus muscle pain11/9/2021neuroscience research institute, peking university7animal models of pain models of chronic inflammatory pain adjuvant-induced arthritis unilateral arthritis inflammation of a hollow viscus ureteral calculosis11/9/2021neuroscience research institute, peking university8扭體模型

6、 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(jiàn)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) 該模型可以模擬腹腔炎癥引起的腹痛癥狀11/9/2021neuroscience research institute, peking university9the abdominal constriction (writhing) test tonic inflammatory pain spinally mediated v

7、isceral/subcutaneous0.9% acetic acid(10 ml/kg; intraperitoneal)11/9/2021neuroscience research institute, peking university10白陶土-鹿角菜膠炎癥模型 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(guò)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

8、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(guò)程一般在第一次注射后2小時(shí)內(nèi)開(kāi)始。動(dòng)物后足紅腫，皮溫升高，pwt值降低等類(lèi)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11/9/2021neuroscience research institute, peking university11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

9、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) 11/9/2021neuroscience research institute, peking university12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11/9/2021neuroscience research institute, pe

10、king university13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(jué)過(guò)敏(hyperalgesia): 對(duì)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).11/9/2021neuroscience research institute, peking university14炎癥痛模型 inflammatory pain

11、 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?chē)啄Ｐ?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?chē)植拷M織的炎癥反應(yīng) 單發(fā)性佐劑關(guān)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(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11/9/2021neuroscience research institute, peking university15ankle joint:intra-articular injection of cfa

12、week 1: acute periodweek 2-3: subacute periodweek 4-9: chronic periodchronic inflammatory pain model - monoarthritis11/9/2021neuroscience research institute, peking university16012345690246810#/*scores of extension pain test#/time (weeks after injection of cfa)ifa: incomplete freunds adjuvantcfa: co

13、mplete freunds adjuvantn=10 / group*p0.05, *p0.01, *p0.001 compared with ifa group#p0.05, #p0.01, #p0.001 compared with left anklechronic inflammatory pain model-monoarthritisifa leftcfa left ifa rightcfa right11/9/2021neuroscience research institute, peking university17animal models of pain neuropa

14、thic pain models experimental anesthesia dolorosa experimental models of painful peripheral neuropathy due to traumatic, partial nerve damage chronic constriction injury partial nerve transection injury spinal nerve transection injury experimental models of painful diabetic neuropathy chemotherapy-e

15、voked painful peripheral neuropathy11/9/2021neuroscience research institute, peking university18neuropathic pain from nerve inflammation eliav 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 i

16、s saturated with cfa. within 24 and 48 h the animals develop heat-hyperalgesia, mechano-hyperalgesia, mechano-allodynia, and (to a lesser 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 experimen

17、tal neuritis of the rat sciatic nerve. pain 1999; 83:169)11/9/2021neuroscience research institute, peking university19l2l3l4l5l6l2l3l4l5chungscciseltzer11/9/2021neuroscience research institute, peking university2011/9/2021neuroscience research institute, peking university21allodynia in rats infected

18、 with varicella zoster virusa small animalmodel for post-herpetic neuralgiafollowing vzv infection of the left footpad rats develop a chronic 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 reproduci

19、ble with animals consistently developing allodynia by 3 days post-infection and continuing to present with symptoms for at least 30 days. 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 investig

20、ate the pathogenic mechanisms underlying the development of vzv-induced allodynia. comparative studies using hsv-1 show that the induction of the chronic allodynia is vzv-specific and is not a result is of virus replication-induced tissue damage or accompanying inflammation.11/9/2021neuroscience res

21、earch institute, peking university22fig. 1. duration of vzv-induced allodynia11/9/2021neuroscience research institute, peking university23fig. 2. reproducibility of the modelthe mean withdrawal thresholds observed in four individual vzv studies (n=24) are presented individually (, ,). the data from

22、the controls (n=24) from these four studieswere pooled and are plotted as a single line ().11/9/2021neuroscience research institute, peking university24fig. 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-

23、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 treated 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 determi

24、ned ipsilateral paws and plotted against time post-infection in days for each group and sem shown. hsv-1 (), hsv plus valaciclovir (), control (). (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

25、 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-infection. 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 show

26、n. vzv (), vzv plus valaciclovir (), control (). the line above the graphs indicates the duration of administration of valaciclovir.11/9/2021neuroscience research institute, peking university25animal models of pain visceral pain models colonic-rectal distension (crd) small bowel distension artificia

27、l kidney stones urinary bladder distension urinary bladder irritants ischemic stimuli (coronary artery occlusion)11/9/2021neuroscience research institute, peking university26chemotherapy-evoked painful peripheral neuropathy (1) painful peripheral neuropathy is a common, although seldom acknowledged,

28、 side effect of cancer chemotherapy. chemotherapy-evoked neuropathic pain has been made using vincristine and paclitaxel. the use of dose that are 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 produ

29、ced a significant mechano-hyperalgesia beginning around the time of the last injection on day 10 and continuing for at least 12 days after dosing ceased. both doses produced a significantly increased threshold to heat-evoked pain. (aley ko, et al. vincristine hyperalgesia in the rat: a model of pain

30、ful cincristine neuropathy in humans, neuroscience 1996; 73: 259)11/9/2021neuroscience research institute, peking university27chemotherapy-evoked painful peripheral neuropathy (2) polomano et al described a paclitaxel-evoked painful peripheral neuropathy in the rat that is not associated with any ev

31、idence of injury to sensory or motor axons and that is not accompanied by significant effects on the animals general health. rats were treated with 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-hyper

32、algesia, mechano-allodynia, and cold-allodynia. the abnormal pain sensations began within several days of the initiation of treatment and lasted for at least several weeks afterward. (polomano rc, et al. a painful peripheral neuropathy in the rat produced by the chemotherapeutic drug, paclitaxel. pa

33、in 2001; 94: 293-304)11/9/2021neuroscience research institute, peking university28colonic-rectal distension in rats, a flexible latex balloon fixed 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 late

34、x 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 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 sil

35、k suture. (gebhart gf, et al. evaluation of visceral pain, in methods in gastrointestinal pharmacology, gaginella, ts ed, crc press, boca ratom 1996, 359)11/9/2021neuroscience research institute, peking university29animal models of pain models of cancer pain 大鼠脛骨乳腺癌痛模型 小鼠足底癌痛模型11/9/2021neuroscience

36、research institute, peking university30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(jué)過(guò)敏、痛覺(jué)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11/9/2021neuroscience research institute, peking university31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 old b16-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)：

37、 0.1m pbs 20ul 對(duì)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, 200211/9/2021neuroscience research institute, peking university32小鼠腳掌腫瘤生長(zhǎng)情況小鼠腳掌腫瘤生長(zhǎng)情況11/9/2021neuroscience research institute, peking university33疼痛的常見(jiàn)癥狀 人類(lèi)的“疼痛”與動(dòng)物的“傷害性感受” 常見(jiàn)癥狀：主要包括on

38、going pain and stimulus-evoked pain 自發(fā)痛(ongoing pain ) 誘發(fā)痛(stimulus-evoked pain)，包括痛覺(jué)過(guò)敏hyperalgesia和痛覺(jué)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，通過(guò)觀察動(dòng)物的行為，實(shí)驗(yàn)者來(lái)推測(cè)動(dòng)物是否發(fā)生了“疼痛”11/9/2021neuroscience research institute, peking university34慢性痛的常見(jiàn)癥狀 自發(fā)痛spontaneous pain 持續(xù)存在的通感覺(jué) 痛覺(jué)過(guò)敏hyperalgesia 弱的痛刺激

39、引起強(qiáng)的痛感覺(jué) 痛覺(jué)超敏allodynia，或稱觸誘發(fā)痛 非痛刺激引起痛感覺(jué)11/9/2021neuroscience research institute, peking university35痛敏的種類(lèi)與機(jī)制 痛敏的種類(lèi)(types of hyperalgesia) 痛敏包括痛覺(jué)過(guò)敏(hyperalgesia)與痛覺(jué)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?chē)茄装Y區(qū)也發(fā)生痛敏11/9/2021neuroscience research institute, peking univers

40、ity36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(jué)超敏 (觸痛)11/9/2021neuroscience research institute, peking university37philosophy of measuring pain the human subject can report his sensations to us. he does so with an act, some sort of behavior- the spoken word, a pen

41、cil 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 private subjective experience (a “sharper” image) is the same as what he would experience under the same circumstances. 11/9/2021neuroscience research

42、institute, peking university38philosophy of measuring pain we assume that other people see like us because they look like us. rats do not look like us. 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 an

43、d depends on exactly what kind of experience we are discussing.11/9/2021neuroscience research institute, peking university39philosophy of measuring pain we 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 i

44、s 45c under normal circumstances, the sensation of pain is tightly related to tissue damage. it is reasonable to argue that this relationship has obvious evolutionary value. it is also an obviously primitive relationship that is likely to be highly conserved in man, rat, other mammals, and probably

45、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 order of the potency of opioids is the same as in human beings and rats.11/9/2021neuroscience research institute, peking university40measuring pain i

46、n animals acute and chronic pain the distinction is arbitrary “acute” refers to pain that lasts for seconds to about a day “chronic” refers to pain that lasts 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 model

47、 but 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 opioid-like receptors.11/9/2021neuroscience research institute, peking university41methods in pain research behavioral: hot (cold) plate, von

48、 frey hair, pain score pharmacological: antagonist, radio ligand binding assay psychological neurochemical: neurotransmitter content measurement with high performance liquid chromatography (hplc) cellular, molecular, and genetic morphological: histochemical, immunohistochemical, fluorescent electrop

49、hysiological: extracellular, multi-channel recording patch clamp evoked potential non-invasive: pet, fmricombination of methods at different levels, integration of above methods11/9/2021neuroscience research institute, peking university42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)

50、eric r. kandel, et al. principles of neuroscience4th edition. fig. 11-8.patch clamp 技術(shù)11/9/2021neuroscience research institute, peking university43影響傷害性感受測(cè)量的因素 動(dòng)物種類(lèi)、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(duì)閾值和反應(yīng)的影響 皮膚基礎(chǔ)溫度對(duì)傷害性熱刺激閾值得影響11/9/2021neuroscience research institute, peking univ

51、ersity44differences of basal thresholds amonginbred strains of mice129aakr b6b10 b/c c3h c58 riiis sm23456n=46-80strainsbasal threshold (s)sex difference of basal thresholdsamong inbred strains129aakrb6b10b/cc3hc58riiissm23456n=14-51strainsbasal threshold (s)strain and sex differences in basal thres

52、hold in mice11/9/2021neuroscience research institute, peking university45測(cè)量疼痛的兩類(lèi)方法 第一類(lèi)：測(cè)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(cè)定刺激的強(qiáng)度和時(shí)程。 第二類(lèi)：測(cè)量產(chǎn)生傷害性反應(yīng)所需的刺激強(qiáng)度和時(shí)程。即刺激是標(biāo)準(zhǔn)化的。與第一類(lèi)不同，它測(cè)定的不是閾值，而是反應(yīng)的大小。11/9/2021neuroscience research institute, peking university46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11/9/2021ne

53、uroscience research institute, peking university47實(shí)驗(yàn)動(dòng)物的疼痛評(píng)價(jià)方法 較理想的行為學(xué)評(píng)價(jià)方法應(yīng)該具備 能區(qū)分動(dòng)物對(duì)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(cè)得的行為改變可以反映動(dòng)物的痛感受 動(dòng)物的行為反應(yīng)對(duì)鎮(zhèn)痛藥物的處理敏感 能將非感覺(jué)性變化，如注意力、活動(dòng)能力等與感覺(jué)性變化區(qū)分開(kāi) 反復(fù)刺激不引起或只引起極小的組織損傷11/9/2021neuroscience research institute, peking university48實(shí)驗(yàn)動(dòng)物的疼痛評(píng)價(jià)方法 簡(jiǎn)單的反射行為 甩尾實(shí)驗(yàn)(t

54、ail flick test) 鉀離子測(cè)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(cè)痛法(hot plate test) 冷板測(cè)痛法(cold plate test) 翻滾實(shí)驗(yàn)(wrotjomg 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)11/9/2021neuroscience research

55、institute, peking university49動(dòng)物疼痛的行為學(xué)研究方法外周神經(jīng)損傷后的機(jī)械痛敏oxoxox11/9/2021neuroscience research institute, peking university51cold-induced ongoing pain after peripheral nerve injury5 c11/9/2021neuroscience research institute, peking university52輻射熱甩尾測(cè)定痛閾（電針）11/9/2021neuroscience research institute, pekin

56、g university5311/9/2021neuroscience research institute, peking university5411/9/2021neuroscience research institute, peking university5511/9/2021neuroscience research institute, peking university56012345690246810#/*scores of extension pain test#/time (weeks after injection of cfa)ifa: incomplete fre

57、unds 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 anklechronic inflammatory pain model-monoarthritisifa leftcfa left ifa rightcfa right11/9/2021neuroscience research institute, peking university5711/9/2021

58、neuroscience research institute, peking university5811/9/2021neuroscience research institute, peking university5911/9/2021neuroscience research institute, peking university6011/9/2021neuroscience research institute, peking university6111/9/2021neuroscience research institute, peking university62the

59、abdominal constriction (writhing) test tonic inflammatory pain spinally mediated visceral/subcutaneous0.9% acetic acid(10 ml/kg; intraperitoneal)11/9/2021neuroscience research institute, peking university6311/9/2021neuroscience research institute, peking university6411/9/2021neuroscience research in

60、stitute, peking university65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)、對(duì)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) 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(jué)超敏痛敏，通覺(jué)超敏 （allodynia)