本文選題：電針 + 慢性神經(jīng)痛��； 參考：《中國(guó)中醫(yī)科學(xué)院》2017年碩士論文

【摘要】：研究背景疼痛是一種與組織損傷相關(guān)聯(lián)的不愉快的感覺(jué)和情緒體驗(yàn),含有感覺(jué)、情緒、認(rèn)知等多個(gè)維度。近年來(lái),有關(guān)痛情緒及痛認(rèn)知障礙的研究受到了越來(lái)越多的關(guān)注。眾所周知:邊緣系統(tǒng)參與腦記憶、感覺(jué)和情緒活動(dòng)。杏仁核是邊緣系統(tǒng)的一個(gè)重要核團(tuán),被稱(chēng)作“情緒腦”,是腦內(nèi)調(diào)節(jié)與焦慮、恐懼相關(guān)行為、自主活動(dòng)和激素水平等生理反應(yīng)的關(guān)鍵結(jié)構(gòu)。慢性疼痛除了疼痛過(guò)敏、認(rèn)知、精神心理的變化外,還伴有大腦結(jié)構(gòu)和功能的改變。近年來(lái)的研究結(jié)果表明:慢性痛可導(dǎo)致腦結(jié)構(gòu)及功能的可塑性變化,腦解剖學(xué)結(jié)構(gòu)重組,腦皮層灰質(zhì)密度的改變。慢性痛中杏仁核突觸可塑性功能發(fā)生變化可能與突觸前后蛋白變化有關(guān),谷氨酸是神經(jīng)系統(tǒng)中主要的興奮性神經(jīng)遞質(zhì),丫-氨基丁酸(GABA)是中樞神經(jīng)系統(tǒng)主要的抑制性神經(jīng)遞質(zhì),突觸后密度蛋白95(PSD-95)是突觸后致密物中的重要結(jié)構(gòu)蛋白,可將位于細(xì)胞膜上的谷氨酸N-甲基-D-天冬氨酸受體(NMDAR)和位于細(xì)胞內(nèi)的信號(hào)轉(zhuǎn)導(dǎo)系統(tǒng)聯(lián)系起來(lái),突觸前基質(zhì)蛋白Piccolo蛋白位于突觸前細(xì)胞質(zhì)基質(zhì)活性區(qū),在大腦神經(jīng)遞質(zhì)傳遞中有重要作用。針刺作為緩解慢性痛的有效且方便快捷的治療方法,在臨床上療效顯著。隨著對(duì)痛情緒在慢性痛發(fā)展過(guò)程中重要性的研究,發(fā)現(xiàn)針刺可以治療疼痛誘發(fā)的情緒改變,針刺可以緩解抑郁、焦慮、失眠等情緒障礙性疾病以及疼痛誘發(fā)的負(fù)性情緒。我們前期研究觀(guān)察到:重復(fù)電針改善慢性神經(jīng)痛性負(fù)性情緒大鼠疼痛的感覺(jué)成分和痛情緒成分的作用可能分別與其上調(diào)杏仁核促皮質(zhì)釋放激素受體(CRF)-1R、(CRF)-2R、谷氨酸N-甲基-D-門(mén)冬氨酸(NMDA)受體亞型NR2B、γ-氨基丁酸(GABA)受體亞型A和C基因表達(dá)和阿片μ受體(MOR)蛋白表達(dá),抑制谷氨酸受體GluA1的降低有關(guān),但其機(jī)制仍遠(yuǎn)不清楚。本研究擬在同一慢性疼痛負(fù)性情緒模型上,觀(guān)察電針“足三里”-“陽(yáng)陵泉”對(duì)動(dòng)物痛行為的感覺(jué)成分和情感成分的影響,它與杏仁核神經(jīng)細(xì)胞突觸可塑性相關(guān)分子GABA受體亞型A β 2,B1,Glu-NMDA受體亞型NR1,突觸蛋白PSD95及Piccolo表達(dá)變化的關(guān)系;杏仁核內(nèi)微量注射GABAA及GABAB受體拮抗劑,驗(yàn)證它們參與電針改善動(dòng)物痛感覺(jué)成分和痛情感成分效應(yīng)的情況,并用透射電鏡觀(guān)察杏仁核細(xì)胞突觸超微結(jié)構(gòu)的變化,試圖從結(jié)構(gòu)和功能相結(jié)合的角度,探討杏仁核介導(dǎo)的針刺緩解慢性痛的作用機(jī)制。材料與方法第一部分實(shí)驗(yàn):健康雄性Wistar大鼠56只(230g-250g),隨機(jī)分為4組:正常組,CCI+NA(chronic compressive injury+NA)模型組,CCI+NA+ 電針(EA)組,CCI+NA+麻醉電針(AEA)組,其中每組8只用于Real-timePCR,其余6只用于Western Blot檢測(cè)。除正常組外,其余各組大鼠行左下肢坐骨神經(jīng)結(jié)扎術(shù),恢復(fù)三天后進(jìn)行足底電刺激造成負(fù)性情緒模型。電針組和麻醉電針組電針刺激雙側(cè)“足三里”、“陽(yáng)陵泉”,1mA,2/15Hz,每天1次,每次30分鐘,共7天。電針干預(yù)后,即刻將放入條件控制箱20分鐘,大于兩小時(shí)后,放于非條件控制箱20分鐘。采用痛覺(jué)測(cè)定儀測(cè)定大鼠雙側(cè)足底熱輻射痛閾值,取雙側(cè)縮足潛伏期差值(PWLD),采用條件性位置偏惡系統(tǒng)測(cè)定痛情緒方面。動(dòng)物直接取材后分別采用Real-time PCR、Western Blot檢測(cè)杏仁核內(nèi)突觸可塑性相關(guān)分子GABAAβ2,GABAB,,Glu-NMDA-NR1,PSD95 及 Piccolo 基因及蛋白表達(dá)。第二部分實(shí)驗(yàn):健康雄性Wistar大鼠50只(230g-250g),隨機(jī)分成5組:正常組,CCI+NA+生理鹽水組,CCI+NA+EA+GABAA抑制劑(Flumazenil)組,CCI+NA+EA+GABAB抑制劑(Phaclofen)組,CCI+NA+EA+DMSO溶劑組,每組10只(數(shù)據(jù)完整者6只)。正常組除外,將其余各組大鼠左下肢坐骨神經(jīng)結(jié)扎�；謴�(fù)5天后,除正常組外,余下4組在腦立體定位儀上實(shí)施雙邊杏仁核埋管(坐標(biāo):P:2.5mm,L,R:4.5mm,H:7.7mm),7天后造負(fù)性情緒模型,雙側(cè)分別微量注射生理鹽水0.5 μL/次/側(cè)、GABAA抑制劑 Flumazenil(2 μ g/μ L)0.5 u L/次/側(cè)、GABAB抑制劑(Phaclofen)(2μ g/μ L)0.5 μ L/次/側(cè)、DMS0 溶劑(5 u g/μ L)0.5 μ L/次/側(cè),推進(jìn)速度 0.1 μL/min,注射完成之后,將大鼠固定于鼠架上電針刺激雙側(cè)“足三里-陽(yáng)陵泉”參數(shù)為1mA,2/15Hz,每天1次,每次30分鐘,共7天。電針結(jié)束后,測(cè)定大鼠雙側(cè)足底熱輻射痛閾值,計(jì)算雙側(cè)縮足潛伏期差值(PWLD)及大鼠在條件控制箱停留時(shí)間。第三部分實(shí)驗(yàn):健康雄性Wistar大鼠16只(230g-250g),實(shí)驗(yàn)動(dòng)物分組、模型制備、電針和麻醉電針干預(yù)、熱痛閾測(cè)定、條件位置偏惡檢測(cè)同實(shí)驗(yàn)一,用4%多聚甲醛和2%戊二醛混合后灌流后取材,運(yùn)用透射電鏡技術(shù)觀(guān)察杏仁核細(xì)胞突觸界面相關(guān)參數(shù)的變化。每個(gè)樣本任選15個(gè)視野拍照。每個(gè)視野任選1個(gè)突觸前、后膜及突觸囊泡較為清晰的突觸,采用圖像分析軟件OPTPro3000。統(tǒng)計(jì)學(xué)處理:所得實(shí)驗(yàn)數(shù)據(jù)為計(jì)量資料,以均數(shù)±標(biāo)準(zhǔn)差(M±SD)表示,用SPSS16.0統(tǒng)計(jì)軟件進(jìn)行數(shù)據(jù)處理,采用重復(fù)性單因素方差分析(行為學(xué)數(shù)據(jù)采用重復(fù)測(cè)量方差分析),組間兩兩比較選用LSD檢驗(yàn),P0.05表示差異有統(tǒng)計(jì)學(xué)意義。實(shí)驗(yàn)結(jié)果:1電針對(duì)慢性痛大鼠痛感覺(jué)和情緒成分行為反應(yīng)的影響造模后(CCI+NA),除正常組外,其余各組大鼠PWLD顯著增加(P0.001),在條件控制箱停留時(shí)間顯著縮短(P0.001),提示:動(dòng)物痛閾降低,負(fù)性情緒產(chǎn)生。與模型組比較,電針和麻醉電針3天和7天后PWLD顯著降低(P0.05),在條件控制箱停留時(shí)間顯著延長(zhǎng)(P0.05)。電針組與麻醉電針組相比,兩組PWLD差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),而麻醉電針3天后條件控制箱停留時(shí)間明顯縮短(P0.05),7天后兩組條件控制箱停留時(shí)間差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),提示電針改善了痛反應(yīng)的感覺(jué)成分和負(fù)性情緒成分。2電針對(duì)杏仁核突觸可塑性相關(guān)分子GABAAβ2,GABAB1,Glu-NMDA-NR1,PSD95及Piccolo基因及蛋白表達(dá)的影響定量RT-PCR及Western Blot的結(jié)果表明,造模后模型組杏仁核內(nèi)GABAAβ2,GABAB1,PSD95基因及蛋白表達(dá)顯著降低(P0.05),Piccolo基因表達(dá)水平顯著降低(P0.05),谷氨酸NMDA-NR1基因表達(dá)有上升趨勢(shì)(P0.05),其蛋白表達(dá)明顯上升(P0.05)。電針后,與模型組相比,電針組NMDA-NR1、GABAAβ2,,GABAB1,PSD95基因及蛋白表達(dá)明顯升高(P0.05),Piccolo基因表達(dá)明顯上升(P0.05);而麻醉電針組NMDA-NR1、GABAAβ2,GABAB1、PSD95基因表達(dá)水平較模型組顯著上調(diào)(P0.05),Piccolo基因表達(dá)明顯下降(P0.001)。這些結(jié)果提示:NMDA-NR1,GABAAβ2,GABAB1,PSD95,Piccolo可能參與針刺改善慢性神經(jīng)痛感覺(jué)的成分,Piccolo有可能還參與電針改善慢性痛負(fù)性情緒的作用。3杏仁核內(nèi)注射GABAA和GABAB受體拮抗劑驗(yàn)證其參與針刺改善痛感覺(jué)和/或痛情緒的作用造模后,除正常組外,其余各組大鼠PWLD值顯著增加(P0.001),電針3天后,與模型組相比,DMS0溶劑對(duì)照電針組PWLD明顯縮短(P0.001),提示該溶劑不影響電針改善痛感覺(jué)成分的效應(yīng),其余各組未見(jiàn)明顯變化(P0.05);與GABAA抑制劑電針組、GABAB抑制劑電針組相比較,DMSO溶劑對(duì)照電針組PWLD明顯縮短(P0.01)。電針7天后,與模型組相比,DMSO溶劑對(duì)照電針組PWLD明顯縮短(P0.001),GABAA抑制劑電針組、GABAB抑制劑電針組有下降趨勢(shì),但無(wú)統(tǒng)計(jì)學(xué)差異。GABAA抑制劑電針組、GABAb抑制劑電針組組間無(wú)統(tǒng)計(jì)學(xué)差異。即說(shuō)明杏仁核內(nèi)注射GABAA和GABAB抑制劑,使電針的鎮(zhèn)痛效應(yīng)明顯減弱。造模后,各組大鼠在條件控制箱停留時(shí)間顯著減少(P0.001),電針3天后,與模型組相比,DMSO溶劑電針組在條件控制箱停留時(shí)間顯著增加(P0.01),提示該溶劑不影響電針改善痛情緒成分的效應(yīng),其余各組未見(jiàn)明顯變化(P0.05)。與GABAA抑制劑電針組、GABAB抑制劑電針組相比較,DMSO電針組在條件控制箱停留時(shí)間顯著增加(P0.01)。電針7天后,與模型組相比,DMSO電針組在條件控制箱停留時(shí)間顯著增加(P0.001),GABAA抑制劑電針組、GABAb抑制劑電針組有升高趨勢(shì),但無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。GABAA抑制劑電針組、GABAb抑制劑電針組組間無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。說(shuō)明在杏仁核內(nèi)阻斷GABAA、GABAb受體后,削弱了電針改善模型大鼠痛情緒的作用。上述結(jié)果表明:GABAA和GABAB均參與電針改善慢性神經(jīng)痛負(fù)性情緒大鼠的痛感覺(jué)及痛情緒成分的效應(yīng)。4電針對(duì)慢性痛負(fù)性情緒大鼠杏仁核突觸結(jié)構(gòu)變化的影響與正常組比,模型組大鼠杏仁核突觸活性帶長(zhǎng)度、突觸后致密物厚度和突觸界面曲率明顯減少(P0.001),突觸間隙明顯增寬(P0.001);與模型組比,電針以及麻醉電針后,大鼠杏仁核突觸活性帶長(zhǎng)度明顯增加(P0.001),突觸后致密物厚度明顯增加(P0.001),突觸間隙寬度明顯減少(P0.001,P0.01),電針組突觸界面曲率增大(P0.05),麻醉電針組突觸界面曲率無(wú)明顯變化(P0.05);電針組與麻醉電針組杏仁核突觸界面參數(shù)變化無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。說(shuō)明電針可明顯改善慢性神經(jīng)痛負(fù)性情緒大鼠杏仁核突觸間隙的增寬,突觸前活性帶的長(zhǎng)度、突觸后致密物(PSD)厚度及突觸界面的曲率的減低。小結(jié)1、電針“足三里-陽(yáng)陵泉”可以改善神經(jīng)病理性慢性痛負(fù)性情緒大鼠痛的感覺(jué)成分和情感成分的行為學(xué)反應(yīng)。2、電針改善痛感覺(jué)成分的作用可能與上調(diào)大鼠杏仁核NMDA-NR1、GABAAβ2,GABAb1、PSD95基因及蛋白表達(dá)水平及Piccolo基因表達(dá)水平有關(guān),電針改善痛情緒成分的作用可能與上調(diào)大鼠杏仁核Piccolo基因表達(dá)水平有關(guān)。3、阻斷杏仁核內(nèi)GABAA以及GABAB后,電針的鎮(zhèn)痛效應(yīng)以及改善痛情緒的作用明顯減弱,GABAA和GABAb受體可能同時(shí)參與了痛感覺(jué)成分與情感成分的調(diào)節(jié)。4、電針可以改善慢性痛引發(fā)的杏仁核神經(jīng)細(xì)胞突觸可塑性變化,調(diào)節(jié)杏仁核突觸界面,明顯減小突觸間隙,增加突觸前活性帶的長(zhǎng)度、突觸后致密物(PSD)厚度及突觸界面的曲率。其改善突觸可塑性可能與其調(diào)節(jié)突觸可塑性相關(guān)蛋白的表達(dá)有關(guān)。
[Abstract]:Background pain is a kind of unpleasant sensation and emotional experience associated with tissue damage, including sensory, emotional, cognitive and other dimensions. In recent years, more and more attention has been paid to the research on pain and cognitive impairment. It is known that the marginal system is involved in brain memory, sensory and emotional activity. The amygdala is a marginal system. An important nuclear group, known as the "emotional brain", is the key structure of physiological responses such as regulation and anxiety, fear related behavior, autonomic activity, and hormone levels in the brain. Chronic pain is accompanied by changes in pain allergy, cognitive, mental and mental changes, and changes in brain structure and function. The changes in the plasticity of the brain structure and function, the reorganization of the brain anatomy, the change in the density of the gray matter in the cortex. The changes in the synaptic plasticity in the amygdala may be related to the changes in the pre and post synaptic proteins in the chronic pain. Glutamate is the main excitatory neurotransmitter in the nervous system, and GABA is the main central nervous system. The inhibitory neurotransmitter, postsynaptic density protein 95 (PSD-95) is an important structural protein in the postsynaptic densification. It can connect the glutamic acid N- methyl -D- aspartic acid receptor (NMDAR) on the cell membrane and the signal transduction system in the cell. The presynaptic protein Piccolo protein is located in the pre synaptic cytoplasmic matrix activity. Areas have an important role in the transmission of neurotransmitters in the brain. Acupuncture as an effective and convenient and quick treatment to alleviate chronic pain has a significant clinical effect. With the study of the importance of pain mood in the process of chronic pain development, acupuncture can be used to treat pain induced mood changes, and acupuncture can relieve depression, anxiety, insomnia and so on. The effect of repeated electroacupuncture on the improvement of the sensory and emotional components of pain in chronic neuropathic negative emotional rats may be compared with the up regulation of the amygdalotropin receptor (CRF) -1R, (CRF) -2R, and N- methyl -D- aspartate (NMD), respectively. A) receptor subtype NR2B, gamma aminobutyric acid (GABA) receptor subtype A and C gene expression and opioid receptor (MOR) protein expression, inhibit the decrease of glutamate receptor GluA1, but its mechanism is still far from clear. This study is to observe the feeling of "Zusanli" - "Yang Ling spring" on animal pain behavior in the same chronic negative emotional model. The effects of components and emotional components on the relationship between the synaptic plasticity related molecular GABA receptor subtype A beta 2, B1, Glu-NMDA receptor subtype NR1, the synaptic protein PSD95 and Piccolo expression in the amygdala nerve cells, and the microinjection of GABAA and GABAB receptor antagonists in the amygdala to verify their involvement in electroacupuncture in improving pain sensory components and pain in animals. The changes in the synaptic ultrastructure of the amygdala cells were observed by transmission electron microscopy. The mechanism of amygdala mediated acupuncture for alleviating chronic pain was explored from the angle of combination of structure and function. The first part of the materials and methods: 56 healthy male Wistar rats (230g-250g) were randomly divided into 4 groups: normal group, C The CI+NA (chronic compressive injury+NA) model group, the CCI+NA+ electroacupuncture (EA) group, and the CCI+NA+ anesthesia electroacupuncture (AEA) group, of which 8 were used in Real-timePCR and the other 6 were used for Western Blot detection. Except the normal group, the rest of the rats were ligation of the left lower extremities, and the electrical stimulation of the foot caused the negative emotional model for three days. The acupuncture group and the electroacupuncture group stimulated bilateral "Zusanli", "Yang Ling spring", 1mA, 2/15Hz, 1 times a day, 30 minutes each time, for a total of 7 days. The prognosis of electroacupuncture would be put into the condition control box for 20 minutes, after more than two hours, and placed in the non conditioned control box for 20 minutes. The difference in the incubation period (PWLD) and the conditioned position deviation system were used to determine the pain mood. After direct sampling, Real-time PCR and Western Blot were used to detect the synaptic plasticity related molecules GABAA beta 2, GABAB, Glu-NMDA-NR1, PSD95 and Piccolo genes and protein expression in the amygdala. The second part of the experiment: healthy male Wistar rats 50 (230g-250g) were randomly divided into 5 groups: normal group, CCI+NA+ saline group, CCI+NA+EA+GABAA inhibitor (Flumazenil) group, CCI+NA+EA+GABAB inhibitor (Phaclofen) group, CCI+NA+EA+DMSO solvent group, 10 rats in each group (6 patients with complete data). The rest of the left lower extremities were ligated in the rest of the rats. 5 days later, except the normal group, The remaining 4 groups carried out bilateral amygdala buried tube (coordinates: P:2.5mm, L, R:4.5mm, H:7.7mm) on the stereotaxis, and 7 days later, the negative emotional model was made. The bilateral injection of physiological saline was 0.5 mu L/ / side, GABAA inhibitor Flumazenil (2 mu L) 0.5 u L/ / side, GABAB inhibitor (2 mu) 0.5 micron / side, 5 solvent (5) U g/ mu L) 0.5 mu L/ sub / side, propelling speed of 0.1 mu L/min, after the injection completed, the rats were fixed on the rat frame to stimulate bilateral "Zusanli Yang spring" parameters to be 1mA, 2/15Hz, 1 times a day, 30 minutes a time for 7 days. After the end of the electroacupuncture, the threshold of the bilateral foot heat radiation pain was measured, and the difference of the bilateral foot latent period (PWLD) and the large amount were calculated. Rats in the condition control box residence time. Third part experiment: 16 healthy male Wistar rats (230g-250g), experimental animals group, model preparation, electroacupuncture and anaesthesia electroacupuncture intervention, thermal pain threshold determination, condition location deviation detection and experiment one, after mixed with 4% polyformaldehyde and 2% amyl two aldehyde, after perfusion, using transmission electron microscope technique to observe almond Changes in the parameters related to the synaptic interface of the nuclear cells. Each sample selected 15 fields of vision to take pictures. Each field of vision was selected 1 presynaptic presynaptic, posterior membrane and synaptic vesicles with a clearer synapse, using the image analysis software OPTPro3000. statistics processing: the obtained experimental data were measured with a mean number of standard deviation (M + SD), and SPSS16.0 statistical software was used. Data processing, repeated single factor analysis of variance (behavior data using repeated measurement of variance analysis), 22 comparison between groups selected LSD test, P0.05 indicated that the difference was statistically significant. Experimental results: 1 electroacupuncture on the pain sensation and emotional components of chronic pain in rats (CCI+NA), except the normal group, the rest of the other groups The PWLD of rats increased significantly (P0.001), the retention time of the condition control box was significantly shortened (P0.001), suggesting that the pain threshold of the animal decreased and the negative emotion produced. Compared with the model group, the PWLD decreased significantly (P0.05) for 3 days and 7 days after the electroacupuncture and Electroacupuncture (P0.05), and the retention time in the condition control box was significantly prolonged (P0.05). Compared with the electroacupuncture group, the two groups of PWLD were compared with the electroacupuncture group. The difference was not statistically significant (P0.05), but the retention time of the condition control box was significantly shortened (P0.05) after 3 days of anaesthetized electroacupuncture (P0.05), and there was no significant difference in the residence time between the two groups of condition control cases (P0.05) after 7 days, suggesting that electroacupuncture improved the sensory components of the pain response and the negative emotional formation of.2 electroacupuncture on the amygdala synaptic plasticity related molecule GABAA beta 2, GABA The effects of B1, Glu-NMDA-NR1, PSD95 and Piccolo gene and protein expression on the quantitative RT-PCR and Western Blot showed that the expression of GABAA beta 2, GABAB1, PSD95 genes and protein expression in the amygdala was significantly reduced (P0.05), and the expression level of Piccolo genes decreased significantly after the model group. After electroacupuncture, the expression of NMDA-NR1, GABAA beta 2, GABAB1, PSD95 gene and protein expression in the electroacupuncture group increased significantly (P0.05), and the expression of Piccolo gene increased significantly (P0.05) compared with the model group. The expression level of the gene expression in the anesthetic electroacupuncture group was significantly higher than that in the model group. P0.001. These results suggest that NMDA-NR1, GABAA beta 2, GABAB1, PSD95, Piccolo may be involved in acupuncture improvement of chronic neuralgia, and Piccolo may also be involved in the effect of electroacupuncture in improving chronic pain and negative emotions. The.3 amygdala is injected with GABAA and GABAB receptor antagonists to verify their participation in acupuncture to improve pain sensation and / or pain mood. In addition to the normal group, the PWLD value of the rest of the rats increased significantly (P0.001). After 3 days of electroacupuncture, the PWLD in the DMS0 solvent control group was significantly shorter than that in the model group (P0.001), suggesting that the solvent did not affect the effect of Electroacupuncture on the improvement of pain sensory components, and the other groups did not change significantly (P0.05); and GABAA inhibitor electroacupuncture group, GABAB suppression. Compared with the electroacupuncture group, the PWLD in the DMSO solvent control group was significantly shorter (P0.01). Compared with the model group, the PWLD in the electroacupuncture group was significantly shorter than the model group (P0.001). The GABAA inhibitor electroacupuncture group and the GABAB inhibitor electroacupuncture group had a downward trend, but there was no statistical difference between the.GABAA inhibitor electroacupuncture group and the GABAb inhibitor electroacupuncture group. The statistical difference. That means the injection of GABAA and GABAB inhibitors in the amygdala made the analgesic effect of the electroacupuncture significantly weakened. After the model, the residence time of the rats in the condition control box was significantly reduced (P0.001). After 3 days of electroacupuncture, the retention time of the DMSO solvent electroacupuncture group increased significantly (P0.01), suggesting that the solvent was not in shadow. The effect of electroacupuncture improved pain emotion components, and the rest of the other groups did not change significantly (P0.05). Compared with the GABAA inhibitor electroacupuncture group, GABAB inhibitor electroacupuncture group, the retention time of the DMSO electroacupuncture group increased significantly (P0.01). After 7 days of electroacupuncture, the retention time of the DMSO Electroacupuncture group was significantly increased (P0.001), G ABAA inhibitor electroacupuncture group, GABAb inhibitor electroacupuncture group has a rising trend, but there is no statistical difference (P0.05).GABAA inhibitor electroacupuncture group, GABAb inhibitor electroacupuncture group there is no statistical difference between groups (P0.05). Indicating that the blockage of GABAA in the amygdala, GABAb receptor weakens the effect of electroacupuncture to the model rats' pain emotion. The results show that GABAA and GA are: GABAA and GA BAB all participated in the effect of Electroacupuncture on the pain and emotional components of the chronic neuralgia negative emotional rats. The effect of.4 electroacupuncture on the synaptic structural changes in the amygdala of the chronic painful negative emotional rats was compared with the normal group. The length of the synapse active zone in the amygdala, the thickness of the postsynaptic density and the curvature of the synaptic interface decreased significantly (P0.001). The synaptic gap was significantly widened (P0.001), and the synaptic active zone length of the amygdala was significantly increased (P0.001), the thickness of the postsynaptic density increased significantly (P0.001), the synaptic gap width decreased significantly (P0.001, P0.01), the synaptic interface curvature increased (P0.05) in the electroacupuncture group (P0.05), and the synaptic interface curvature of the electroacupuncture group. There was no significant change (P0.05); there was no significant difference in the synaptic interface parameters between the electroacupuncture group and the anaesthetized electroacupuncture group (P0.05), indicating that the electroacupuncture could obviously improve the widening of the synaptic gap in the amygdala, the length of the presynaptic active band, the thickness of the postsynaptic density (PSD) and the curvature of the synapse interface in the chronic neuropathic rats. 1, Electroacupuncture "Zusanli Yang Ling spring" can improve the behavioral response of the sensory and emotional components of neuropathic chronic pain negative emotional rats. The effect of Electroacupuncture on the improvement of pain sensory components may be related to the up-regulation of NMDA-NR1, GABAA beta 2, GABAb1, PSD95 gene, the level of egg white and the level of Piccolo gene expression in the amygdala of rats. The effect of Electroacupuncture on improving the emotional components of pain may be related to the increase of the expression level of Piccolo gene in the amygdala of rats. After blocking the GABAA and GABAB in the amygdala, the analgesic effect of the electroacupuncture and the effect of improving the pain mood are obviously weakened. The GABAA and GABAb receptors may participate in the regulation of.4 in the sense of pain and emotional components at the same time, and the electroacupuncture can be changed. The synaptic plasticity changes in amygdala nerve cells induced by good chronic pain, modulate the synaptic interface of the amygdala, reduce the synaptic gap, increase the length of the presynaptic active band, the thickness of the postsynaptic density (PSD) and the curvature of the synaptic interface. The improvement of synaptic plasticity may be related to the regulation of the expression of synaptic plasticity related protein.
【學(xué)位授予單位】：中國(guó)中醫(yī)科學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R245

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