本文選題：痛情緒 + 條件性位置回避(CPA)��； 參考：《山西醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:疼痛包含兩種成分,即痛的感覺分辨(sensory-discriminative)與痛的情緒反應(yīng)(emotional-affective dimensions)[1]。痛的感覺分辨感受痛刺激的性質(zhì)、強(qiáng)度和位置等屬性;痛的情緒體驗(yàn)包括焦慮、厭惡、恐懼等不良情緒。大量證據(jù)表明,電針刺激可以緩解疼痛的感覺分辨,且電針是通過激活體內(nèi)的阿片受體來發(fā)揮緩解疼痛的作用[2-6]。一些臨床研究和動(dòng)物實(shí)驗(yàn)表明,電針刺激也可以緩解人和動(dòng)物的痛情緒反應(yīng),但作用機(jī)制尚不清楚[7,8]。人類和動(dòng)物研究發(fā)現(xiàn),ACC(anterior cingulate cortex,ACC)的吻側(cè)部(r ACC)在痛情緒中有著重要的作用[9-12]。有實(shí)驗(yàn)證明,痛情緒引起的條件性位置回避(conditioned place avoidance,CPA)反應(yīng)可通過切除大鼠雙側(cè)r ACC得到緩解[13-16]。因此,我們推測(cè)傷害性刺激引起的痛情緒反應(yīng),可能會(huì)通過激活r ACC神經(jīng)元的阿片受體得到緩解。前期實(shí)驗(yàn)中,我們?cè)诖笫髍 ACC內(nèi)給予μ-阿片受體激動(dòng)劑DAMGO,結(jié)果發(fā)現(xiàn)其可緩解由完全弗氏佐劑(complete Freund’s adjuvant,CFA)誘導(dǎo)的條件性位置回避,但不影響大鼠的熱縮足反射潛伏期(paw withdrawal latency,PWL)[17]。由此得出結(jié)論,大鼠r ACC內(nèi)μ-阿片受體的活化緩解了大鼠的痛情緒反應(yīng),且此過程不依賴于大鼠的疼痛感覺分辨而實(shí)現(xiàn)。基于前期實(shí)驗(yàn)結(jié)果,我們進(jìn)一步推測(cè),電針刺激緩解痛情緒反應(yīng)也可能通過r ACC內(nèi)μ-阿片受體介導(dǎo)。實(shí)驗(yàn)過程中,我們先觀察電針刺激是否可以緩解CFA誘導(dǎo)的CPA反應(yīng),然后預(yù)先在大鼠r ACC內(nèi)注射μ-阿片受體拮抗劑CTOP后再給予電針刺激,觀察r ACC內(nèi)μ-阿片受體的拮抗劑是否可反轉(zhuǎn)電針緩解痛情緒的作用,并試圖對(duì)其機(jī)制進(jìn)行探討。方法:1.建立由CFA誘導(dǎo)的持續(xù)性疼痛模型取250-270g成年雄性SD(Sprague Dawley)大鼠,左側(cè)足底皮下注射0.08ml完全弗氏佐劑(Complete Freund’s adjuvant,CFA),建立炎癥性疼痛模型。對(duì)照組為左側(cè)足底皮下注射0.08ml生理鹽水(NS)。2.建立由CFA誘導(dǎo)的條件性位置回避(CPA)模型將250-270g成年雄性SD大鼠,隨機(jī)分為2組:(1)足底注射NS(0.08ml)組;(2)足底注射CFA(0.08ml)組。比較大鼠腳掌注射CFA前后大鼠在痛環(huán)境中停留的時(shí)間,并計(jì)算回避分?jǐn)?shù)(CPA Score),判斷大鼠是否產(chǎn)生CPA反應(yīng)。3.電針刺激取成年雄性SD大鼠250-270g,隨機(jī)分為真電針組(electroacupuncture EA)和假電針組(sham EA)。在腳掌注射NS/CFA后,將針灸針插入大鼠雙側(cè)環(huán)跳穴(GB30),用膠帶固定于大腿。通過導(dǎo)線將針灸針與電針儀相連,通電后開始電針刺激。4.檢測(cè)熱縮足反射潛伏期(Paw Withdrawal Latency,PWL)在CPA的第一天和第三天分別檢測(cè)各組大鼠的PWL,觀察CFA對(duì)大鼠的熱痛行為反應(yīng)的影響,以及給予電針刺激、r ACC注射不同劑量CTOP后大鼠PWL數(shù)值的變化。5.實(shí)驗(yàn)分組根據(jù)左側(cè)足底皮下注射、r ACC區(qū)給藥和雙側(cè)環(huán)跳穴刺激將大鼠分為以下8組。(1)足底注射NS,r ACC注射NS,EA;(2)足底注射NS,r ACC注射NS,sham EA;(3)足底注射CFA,r ACC注射NS,EA;(4)足底注射CFA,r ACC注射NS,sham EA;(5)足底注射NS,r ACC分別注射2/10/20(nmol/μl)CTOP,EA;(6)足底注射NS,r ACC分別注射2/10/20(nmol/μl)CTOP,sham EA;(7)足底注射CFA,r ACC分別注射2/10/20(nmol/μl)CTOP,EA;(8)足底注射CFA,r ACC分別注射2/10/20(nmol/μl)CTOP,sham EA。結(jié)果:1.足底注射CFA可誘導(dǎo)大鼠產(chǎn)生條件性位置回避(CPA)反應(yīng)。足底注射CFA與足底注射NS后相比,大鼠的熱縮足反射潛伏期(PWL)明顯縮短(9.1±1.0 s vs 13.1±1.9 s,P0.05,n=5),足底注射CFA可引起大鼠產(chǎn)生炎癥性疼痛。足底注射NS組,(Post 352.1±113.8 s vs Pre 317.2±67.8 s,P0.05,n=5),大鼠在痛環(huán)境停留時(shí)間第三天和第一天相比沒有顯著性差異。足底注射CFA組,(Post151.4±58.38 s vs Pre 323.5±105.5 s,P0.05,n=6),大鼠在痛環(huán)境停留時(shí)間第三天明顯小于第一天,有統(tǒng)計(jì)學(xué)差異。足底注射CFA組和足底注射NS組回避分?jǐn)?shù)之間有顯著性差別(-172.1±97.3 s vs 12.6±81.2 s,P0.05,n1=5,n2=6),足底注射CFA組回避分?jǐn)?shù)比較大,對(duì)痛環(huán)境產(chǎn)生厭惡反應(yīng)。2.電針刺激可緩解CFA誘導(dǎo)的大鼠條件性位置回避(CPA)反應(yīng)。大鼠左側(cè)足底注射CFA后,在雙側(cè)環(huán)跳穴分別給予真電針刺激(EA)或者假電針刺激(sham EA)。炎性痛大鼠模型中,假電針刺激后,大鼠在痛環(huán)境停留的時(shí)間第三天和第一天相比縮短,有顯著性差異,(Post 151.4±23.8 s vs Pre 323.5±43.1 s,P0.05,n=6);真電針刺激后,大鼠在痛環(huán)境中停留時(shí)間第三天與第一天相比沒有統(tǒng)計(jì)學(xué)差異,(Post 248.9±144.1 s vs Pre 234.9±54.57 s,P0.05,n=5)。真電針組(EA)回避分?jǐn)?shù)明顯小于假電針組(sham EA),兩組有統(tǒng)計(jì)學(xué)差異(13.9±47.2 s vs-172.1±39.7 s,P0.05,EA組n=5,sham EA組n=6)。大鼠左側(cè)足底注射NS后,在雙側(cè)環(huán)跳穴分別給予真電針刺激(EA)或者假電針刺激(sham EA)。兩組大鼠在痛環(huán)境中停留時(shí)間第三天與第一天相比均沒有統(tǒng)計(jì)學(xué)差異,假電針組(Post 352.1±50.9 s vs Pre 317.2±30.3 s,P0.05,n=5);真電針組(Post250.2±112.4 s vs Pre 305.6±45.9 s,P0.05,n=5)。兩組回避分?jǐn)?shù)相比沒有統(tǒng)計(jì)學(xué)差異(-64.49±52.9 s vs 12.6±36.3 s,P0.05,NS+sham EA組n=5,NS+EA組n=9)。電針刺激正常大鼠不會(huì)引起大鼠的厭惡或者獎(jiǎng)賞反應(yīng)。大鼠左側(cè)足底注射CFA后,在雙側(cè)環(huán)跳穴分別給予真電針刺激(EA)或假電針刺激(sham EA),在第二天測(cè)量大鼠的熱痛反應(yīng),PWL之間沒有統(tǒng)計(jì)學(xué)差異(12.0±1.4s vs 9.1±0.4 s,P0.05,n=5),電針刺激雙側(cè)環(huán)跳穴不影響刺激后第二天由CFA引起的熱痛行為。3.r ACC內(nèi)注射CTOP反轉(zhuǎn)了電針刺激緩解痛情緒的作用。足底注射CFA組,r ACC內(nèi)注射μ-阿片受體拮抗劑CTOP后進(jìn)行電針刺激。足底注射CFA組,在r ACC內(nèi)注射NS后進(jìn)行電針刺激,作為對(duì)照組。大鼠在痛環(huán)境中停留時(shí)間第三天與第一天相比:CTOP組(Post 210.5±59.8 s vs Pre 307.8±47.0 s,P0.05,n=5);NS組(Post 248.9±64.5 s vs Pre 234.9±24.4 s,P0.05,n=5)。CTOP組回避分?jǐn)?shù)明顯大于NS組,有顯著性差異(-127.5±24.1 s vs 13.9±47.2 s,P0.05,n=5)。r ACC內(nèi)注射CTOP反轉(zhuǎn)了電針刺激緩解痛情緒的作用。足底注射CFA組,r ACC內(nèi)注射μ-阿片受體的拮抗劑CTOP或NS,不進(jìn)行電針刺激。大鼠在痛環(huán)境中停留時(shí)間第三天與第一天相比:CTOP組(Post 255.1±71.8 s vs Pre 403.5±46.2 s,P0.05,n=5);NS組(Post 151.4±23.8 s vs Pre 323.5±43.1 s,P0.05,n=6);CTOP組和NS組回避分?jǐn)?shù)沒有統(tǒng)計(jì)學(xué)差異(-148.4±50.5 s vs-172.1±39.7 s,P0.05,n=5)。r ACC內(nèi)注射CTOP本身不會(huì)影響大鼠的痛情緒反應(yīng)。r ACC內(nèi)注射2/10/20(nmol/μl)的μ-阿片受體拮抗劑CTOP,反轉(zhuǎn)電針刺激緩解痛情緒的作用效果不同。大鼠在痛環(huán)境中停留時(shí)間第三天與第一天相比:2 nmol/μl CTOP組與NS組之間沒有統(tǒng)計(jì)學(xué)差異(-2.0±18.8 s vs 13.9±47.2 s,P0.05,n=7),10 nmol/μl CTOP組與NS組之間有統(tǒng)計(jì)學(xué)差異(-97.3±30.3 s vs 13.9±47.2 s,P0.05,n=7),20 nmol/μl CTOP組與NS組之間有統(tǒng)計(jì)學(xué)差異(-127.5±24.1 s vs 13.9±47.2 s,P0.05,n=7);10nmol/μl CTOP組與2 nmol/μl CTOP組之間有統(tǒng)計(jì)學(xué)差異(-97.3±30.3s vs-2.0±18.8 s,P0.05,n=7),20nmol/μl CTOP組與2 nmol/μl CTOP組之間有統(tǒng)計(jì)學(xué)差異(-127.5±24.1 s vs-2.0±18.8 s,P0.05,n=7),10nmol/μl CTOP組與20 nmol/μl CTOP組之間沒有統(tǒng)計(jì)學(xué)差異(-97.3±30.3 s vs-127.5±24.1 s,P0.05,n=7)。預(yù)先在r ACC內(nèi)注射2 nmol/μl CTOP未觀察到反轉(zhuǎn)電針緩解痛情緒的作用,預(yù)先在r ACC內(nèi)給予10nmol/μl CTOP則反轉(zhuǎn)了電針緩解痛情緒的作用,預(yù)先在r ACC內(nèi)給予20nmol/μl CTOP反轉(zhuǎn)電針緩解痛情緒的作用更強(qiáng),但與10nmol/μl CTOP的作用相比沒有統(tǒng)計(jì)學(xué)差異。大鼠左側(cè)足底注射CFA,r ACC注射2/10/20(nmol/μl)CTOP的各組大鼠PWL值與r ACC注射NS相比,都沒有統(tǒng)計(jì)學(xué)差異(P0.05)。各組PWL值為:CFA+NS組10.9±0.9 s,n=6;CFA+2nmol/μl CTOP組7.2±0.6 s,n=5;CFA+10nmol/μl CTOP組11.0±1.3 s,n=5;CFA+20nmol/μl CTOP組8.1±2.1 s,n=5。r ACC內(nèi)注射CTOP不改變CFA引起的熱痛行為。結(jié)論:1.電針刺激可緩解CFA誘導(dǎo)的大鼠CPA反應(yīng)(痛情緒)。2.電針刺激緩解CFA誘導(dǎo)的痛情緒的作用是通過激活r ACC內(nèi)μ-阿片受體實(shí)現(xiàn)。
[Abstract]:Objective: the pain consists of two components: the properties, intensity and location of sensory discrimination of [1]. pain in the pain of sensory discrimination (sensory-discriminative) and painful emotional response (emotional-affective dimensions); the emotional experience of pain includes anxiety, dislikes, and fear. The sensory discrimination of pain is alleviated and the electroacupuncture is activated by activating the opioid receptor in the body to play a role in alleviating pain. [2-6]. some clinical studies and animal experiments have shown that electroacupuncture can also relieve the pain and emotional responses of humans and animals, but the mechanism of action is not yet clear in [7,8]. human and animal studies, ACC (anterior cingulate cortex, A) CC (R ACC) plays an important role in the pain mood. [9-12]. has proved that the conditioned place avoidance (conditioned place avoidance, CPA) reaction caused by pain emotion can be relieved by the removal of bilateral R ACC in rats. We speculate that the pain emotional response caused by nociceptive stimuli may be activated by activating R God. The opioid receptor of the trans element was relieved. In the previous experiment, we gave the micron opioid receptor agonist DAMGO in the rat R ACC, and found that it could alleviate the conditional position avoidance induced by the complete Freund's adjuvant (complete Freund 'adjuvant, CFA), but did not affect the latent period of the rat's heat contraction reflex (paw withdrawal latency, PWL). It is concluded that the activation of micron opioid receptor in R ACC in rats relieves the pain response of rats, and this process is not dependent on the pain sensation resolution of rats. Based on the previous experimental results, we further speculate that the electroacupuncture stimulation may also be mediated by the R ACC opioid receptor. Whether the electroacupuncture stimulation can relieve the CPA response induced by CFA, and then injecting the micron opioid receptor antagonist CTOP in the rat R ACC to the electroacupuncture stimulation in advance, observe whether the antagonist of the R opioid receptor in R ACC can reverse the effect of the electroacupuncture on the pain emotion, and tries to explore the mechanism of it. Method: 1. the establishment of CFA induced by CFA The model of 250-270g adult male SD (Sprague Dawley) rats was taken from the persistent pain model, and the left foot was injected subcutaneously with 0.08ml complete Freund's adjuvant (Complete Freund 's adjuvant, CFA), and the inflammatory pain model was established. The control group was subcutaneously injected with 0.08ml physiological saline (NS) to establish a conditioned location avoidance model. The adult male SD rats of 250-270g were randomly divided into 2 groups: (1) the plantar injection of NS (0.08ml) group; (2) the plantar injection of CFA (0.08ml) group. The time of the rats' paws before and after the injection of CFA in the pain environment was compared and the avoidance fraction (CPA Score) was calculated to determine whether the rat generated CPA reaction.3. electroacupuncture stimulation to take adult male SD rats, random. It is divided into the true electroacupuncture group (electroacupuncture EA) and the false electroacupuncture group (sham EA). After the injection of NS/CFA in the foot, the acupuncture needle is inserted into the bilateral ring jump point (GB30) in the rat, and the needle is attached to the thigh with the adhesive tape. The acupuncture needle is connected with the electroacupuncture instrument through the wire. The electroacupuncture stimulates.4. to detect the latent period of the reflex (Paw Withdrawal Latency, PWL) in the CPA. The PWL of rats in each group was detected on the first and third days, and the effect of CFA on the heat pain behavior of rats was observed and the stimulation of electroacupuncture was given. The PWL value of R ACC was given after CTOP in different doses of CTOP. The.5. experiment group was subdivided into 8 groups according to the left foot, and the R ACC district administration and bilateral ring hopping stimulation were divided into the following groups. (1) foot Injection of NS, R ACC injection NS, EA; (2) foot injection NS, R ACC injection NS, sham EA; (3) foot injection CFA. Do not injecting 2/10/20 (nmol/ L) CTOP, EA; (8) CFA in the foot, R ACC injection 2/10/20 (nmol/ Mu L) CTOP, which can induce conditioned place avoidance response in rats. .05, n=5), the plantar injection of CFA could cause inflammatory pain in rats. NS group was injected into the plantar group (Post 352.1 + 113.8 s vs Pre 317.2 + 67.8 s, P0.05, n=5). There was no significant difference between the rats in the pain environment for third days and the first day. There was a significant difference between the third days of the residence time of third days. There were significant differences between the foot injection group and the NS group (-172.1 + 97.3 s vs 12.6 + 81.2 s, P0.05, n1=5, n2=6). The avoidance fraction of the foot injection CFA group was larger, and the negative reaction to the pain ring was relieved by CFA induction. Rat conditioned place avoidance (CPA) reaction. After CFA was injected into the left foot of the rat, real electroacupuncture stimulation (EA) or pseudo electroacupuncture stimulation (sham EA) were given at the bilateral ring jump points. In the model of inflammatory pain rats, the time of the rats in the pain environment was shortened third days compared to the first day, and there was a significant difference (Post 151.4 + 23.8 s VS). Pre 323.5 + 43.1 s, P0.05, n=6); after real electroacupuncture stimulation, there was no statistical difference between the third days in the pain environment and the first day (Post 248.9 + 144.1 s vs Pre 234.9 + 54.57 s, P0.05, n=5). The true electroacupuncture group (EA) avoidance score was significantly smaller than that of the false electroacupuncture group, and the two groups had statistical differences (13.9 + 47.2 S, P0.05, group EA, n=5, sham EA n=6). After injection of NS in the left foot of the rat, the third days of the two groups of rats in the pain environment were not significantly different from those in the first day (sham EA). True electroacupuncture group (Post250.2 + 112.4 s vs Pre 305.6 + 45.9 s, P0.05, n=5). There was no statistical difference between the two groups (-64.49 + 52.9 s vs 12.6 + 36.3 s). No real electroacupuncture stimulation (EA) or pseudo electroacupuncture stimulation (sham EA) was used to measure the heat pain response of rats in second days. There was no statistical difference between PWL (12 + 1.4s vs 9.1 + 0.4 s, P0.05, n=5). Electroacupuncture stimulation of bilateral ring hops did not affect the heat pain caused by CFA second days after stimulation. An injection of CFA in group CFA, R ACC injection of CTOP opioid receptor antagonist CTOP for electroacupuncture stimulation. The plantar injection of CFA group, the injection of NS in R ACC and electroacupuncture stimulation, as the control group. The duration of the stay in the pain environment was third days compared to the first day: CTOP group (Post 210.5 + 59.8 s vs 307.8 + 47 47 The avoidance score of OST 248.9 + 64.5 s vs Pre 234.9 + 24.4 s, P0.05, n=5) was significantly greater than that of the NS group, and there was a significant difference (-127.5 + 24.1 s vs 13.9 + 47.2). Rats in the pain environment were stopped for third days compared with the first day: group CTOP (Post 255.1 + 71.8 s vs Pre 403.5 + 46.2 s, P0.05, n=5), NS group (Post 151.4 + 23.8 s vs 323.5 + 43.1). The body did not affect the pain emotion response of the rats to the.R ACC injection of 2/10/20 (nmol/ Mu L) micron opioid receptor antagonist CTOP, and the effect of the reverse electroacupuncture stimulation on the pain emotion was different. The duration of the stay in the pain environment was third days compared with the first day: there was no statistical difference between the 2 nmol/ Mu L CTOP group and the NS group (-2.0 + 18.8 s 13.9 + 47.2) S, P0.05, n=7), there are statistical differences between the 10 nmol/ L CTOP group and the NS group (-97.3 + 30.3 s vs 13.9 + 47.2 s, P0.05, and P0.05). N=7), there were statistical differences between the 20nmol/ L CTOP group and the 2 nmol/ L CTOP group (-127.5 + 24.1 s vs-2.0 + 18.8 s, P0.05, 24.1). The effect of giving 10nmol/ Mu L CTOP in advance in R ACC reverses the effect of Electroacupuncture on relieving pain emotion, and the effect of giving 20nmol/ u l CTOP reverse electroacupuncture in R ACC to relieve pain emotion is stronger, but there is no statistical difference compared with 10nmol/ micron L. The PWL values of rats were not statistically different from that of R ACC injection NS (P0.05). The PWL values in group CFA+NS were 10.9 + 0.9 s, n=6 and CFA+2nmol/ u l CTOP group 7.2 + 0.6. The effect of CFA induced CPA response (pain).2. electroacupuncture on alleviating CFA induced pain was achieved by activating the R opioid receptor in R ACC.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R402

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相關(guān)期刊論文 前4條

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