本文選題：電針 + 偏頭痛�。� 參考：《首都醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的偏頭痛是常見(jiàn)的原發(fā)性頭痛,多以單側(cè)或雙側(cè)顳部反復(fù)疼痛為主要的臨床表現(xiàn)。針刺在治療偏頭痛的臨床療效確切,但其治療機(jī)制尚未完全明確。隨著醫(yī)學(xué)的不斷進(jìn)步,對(duì)偏頭痛發(fā)病機(jī)制研究從細(xì)胞水平逐漸深入到神經(jīng)肽水平,大量研究[1,2]發(fā)現(xiàn)多種痛性神經(jīng)肽與偏頭痛的發(fā)作密切相關(guān),如神經(jīng)肽家族群SP及其受體NK-1、降鈣素相關(guān)肽(CGRP)、5-羥色胺(5-HT)、一氧化氮(NO)、內(nèi)皮素等,其中P物質(zhì)與降鈣素相關(guān)肽(CGRP)作為神經(jīng)遞質(zhì)在感覺(jué)傳導(dǎo)和痛覺(jué)調(diào)制中作用越來(lái)越引起研究者的注意。有大量實(shí)驗(yàn)[3,4]已證明神經(jīng)肽SP及CGRP在神經(jīng)調(diào)制中既能傳遞痛覺(jué)信息,有致痛作用;又能促進(jìn)腦啡肽(enkehalin,ENK)的釋放,從而達(dá)到鎮(zhèn)痛作用。故本實(shí)驗(yàn)通過(guò)皮下注射硝酸甘油備制模型,觀察電針對(duì)急性發(fā)作期偏頭痛大鼠中腦SP、NK-1分布與表達(dá)及血漿CGRP濃度變化的影響,來(lái)探討電針?lè)乐渭毙园l(fā)作期偏頭痛相關(guān)機(jī)制。方法250±50g健康Wistar雄性大鼠80只,每4只一個(gè)籠子,清潔級(jí)實(shí)驗(yàn)室喂養(yǎng),自由攝食、飲水。飼養(yǎng)溫度(22±1?C),濕度40%~70%,噪音60分貝,換氣次數(shù)10~20次/小時(shí),光照時(shí)間每天12小時(shí)(8:00~20:00),光照時(shí)間模擬晝夜交替,適應(yīng)性飼養(yǎng)7天。隨機(jī)分為4組:空白組、模型組、電針組、偽電針組,每組20只。空白組不作任何處理,正常飲食攝水飼養(yǎng)7天,于第8天皮下注射生理鹽水;模型組不作電針干預(yù)治療,正常飲食攝水飼養(yǎng)7天,于第8天皮下注射硝酸甘油制備偏頭痛大鼠模型;電針組通過(guò)電針雙側(cè)太陽(yáng)、風(fēng)池、太沖、足臨泣穴干預(yù)7天;偽電針組進(jìn)行非穴位(非穴位點(diǎn)為非經(jīng)非穴部位,其位置不固定)電針干預(yù)7天后,于第8天分別對(duì)兩組進(jìn)行皮下注射硝酸甘油制備偏頭痛大鼠模型。造模成功后,對(duì)兩組進(jìn)行相應(yīng)的電針干預(yù)治療。本實(shí)驗(yàn)主要采用以下幾項(xiàng)作為偏頭痛大鼠觀察指標(biāo):撓頭、爬籠、咬尾、往返運(yùn)動(dòng)的次數(shù)并計(jì)數(shù)(每個(gè)癥狀出現(xiàn)1次計(jì)1分);采用酶聯(lián)免疫吸附法(ELISA)測(cè)定大鼠血漿中CGRP的濃度;免疫組化檢測(cè)中腦導(dǎo)水管周圍灰質(zhì)區(qū)SP、NK-1陽(yáng)性細(xì)胞數(shù);通過(guò)實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)大鼠中腦導(dǎo)水管周圍灰質(zhì)區(qū)SP及其受體NK-1基因表達(dá)。結(jié)果1、電針對(duì)偏頭痛大鼠血漿CGRP濃度變化的影響:與空白組比較,模型組、偽電針組血漿CGRP濃度均明顯升高(P0.05),電針組無(wú)明顯變化(P0.05);與模型組比較,電針組濃度明顯降低(P0.05),偽電針組無(wú)明顯變化(P0.05);與偽電針組比較,電針組濃度明顯降低(P0.05)。提示電針能顯著降低急性發(fā)作期偏頭痛大鼠血漿中CGRP濃度,其療效明顯優(yōu)于偽電針治療。2、電針對(duì)大鼠中腦組織SP及NK-1基因表達(dá)影響:與空白組比較,模型組、電針組、偽電針組SP、NK-1m RNA表達(dá)量明顯增高(P0.05);與模型組比較,電針組、偽電針組SP、NK-1m RNA表達(dá)量明顯降低(P0.05);與偽電針組比較,電針組SP、NK-1 m RNA表達(dá)量明顯降低(P0.05)。提示電針能顯著降低急性發(fā)作期偏頭痛PAG區(qū)的SP及NK-1的基因表達(dá)水平,其療效明顯優(yōu)于偽電針治療。3、電針對(duì)大鼠中腦導(dǎo)水管周圍灰質(zhì)區(qū)SP及NK-1蛋白表達(dá)及分布的影響。SP陽(yáng)性細(xì)胞數(shù):與空白組(A組)比較,模型組(B組)、偽電針組(D組)陽(yáng)性細(xì)胞數(shù)均明顯升高(P0.05),電針組(C組)無(wú)明顯變化(P0.05);與模型組(B組)比較,電針組(C組)、偽電針組(D組)陽(yáng)性細(xì)胞數(shù)均明顯降低(P0.05);與偽電針組(D組)相比較,電針組(C組)陽(yáng)性細(xì)胞數(shù)明顯降低(P0.05)。NK-1陽(yáng)性細(xì)胞數(shù)表達(dá):與空白組(A組)比較,模型組(B組)、電針組(C組)、偽電針組(D組)均明顯升高,且差異有統(tǒng)計(jì)學(xué)意義(P0.05);模型組(B組)、電針組(C組)、偽電針組(D組)三組間各自比較均無(wú)明顯差異(P0.05)。提示,電針能顯著降低急性發(fā)作期偏頭痛大鼠中腦SP的陽(yáng)性分布及表達(dá),其療效明顯優(yōu)于偽電針治療;對(duì)于NK-1陽(yáng)性表達(dá)及分布的影響有降低趨勢(shì),但無(wú)明顯變化。結(jié)論 電針通過(guò)下調(diào)急性發(fā)作期偏頭痛大鼠痛性相關(guān)神經(jīng)肽CGRP、SP表達(dá)量,發(fā)揮防治偏頭痛的作用。
[Abstract]:Objective migraine is a common primary headache, mostly with unilateral or bilateral temporal and recurrent pain as the main clinical manifestation. Acupuncture is effective in the treatment of migraine, but its mechanism of treatment is not completely clear. With the continuous progress of medicine, the study of the pathogenesis of migraine from cell level to neuropeptide level, [1,2] found that a variety of painful neuropeptides are closely related to the attack of migraine, such as the neuropeptide family group SP and its receptor NK-1, calcitonin related peptide (CGRP), 5- serotonin (5-HT), nitric oxide (NO), endothelin, etc., in which P substance and calcitonin related peptide (CGRP) Act more and more as neurotransmitters in sensory conduction and pain modulation Researchers' attention. A large number of experiments [3,4] have proved that neuropeptide SP and CGRP can not only transmit pain information, cause pain, but also promote the release of enkephalin (enkehalin, ENK) and thus achieve analgesic effect. Therefore, the experiment was made by subcutaneous injection of nitroglycerin to observe the effect of Electroacupuncture on acute migraine rats. The influence of the distribution and expression of SP, NK-1 and the change of CGRP concentration in the plasma to discuss the mechanism of electroacupuncture in the prevention and treatment of acute migraine. Methods 80 healthy Wistar male rats of 250 + 50g, every 4 cages, free feeding, drinking water, feeding temperature (22 + 1? C), humidity 40%~70%, noise 60 decibels, air exchange times 10~20 The light time was 12 hours a day (8:00~20:00), and the light time was simulated day and night and adapted for 7 days. It was randomly divided into 4 groups: blank group, model group, electroacupuncture group, pseudo-needle group, 20 in each group. The blank group was not treated with any treatment, normal diet for 7 days and eighth days by subcutaneous injection of saline; the model group did not do electroacupuncture intervention, The rat model of migraine was prepared by subcutaneous injection of nitroglycerin on the eighth day for 7 days, and the electroacupuncture group was treated with Electroacupuncture of the sun, the wind pond, the Tai Chi, and the foot and the weeping point for 7 days. The pseudo electroacupuncture group carried out the non acupoint (non acupoint points for non acupoint points, and its position was not fixed) for 7 days, and the two groups were carried out on the eighth day respectively. The rat model of migraine was prepared by injection of nitroglycerin. After the model was successful, the two groups were treated with electroacupuncture. This experiment mainly used the following indexes as observation indexes of migraine rats: bending head, crawling cage, tail biting, and counting the number of round-trip times (1 times each symptom was 1); enzyme linked immunosorbent assay (ELISA) The concentration of CGRP in the rat plasma; immunohistochemistry to detect the number of SP, NK-1 positive cells in the periaqueductal gray area of the middle cerebral aqueduct; the expression of SP and its receptor NK-1 gene in the periaqueductal gray area of the rat by real-time fluorescence quantitative PCR. Results 1, the effect of Electroacupuncture on the plasma CGRP concentration in the migraine rats: compared with the blank group, the model group, The concentration of plasma CGRP in the pseudo electroacupuncture group increased significantly (P0.05), and there was no obvious change in the electroacupuncture group (P0.05). Compared with the model group, the concentration of electroacupuncture group decreased significantly (P0.05), and there was no obvious change in the pseudo-electroacupuncture group (P0.05), and the concentration of electroacupuncture group decreased significantly (P0.05) compared with the pseudo electroacupuncture group (P0.05). It suggested that electroacupuncture could significantly reduce the CGRP in the plasma of migraine rats at acute attack. The effect was obviously better than that of the pseudo-electroacupuncture treatment of.2, and the effect of Electroacupuncture on the expression of SP and NK-1 gene in the middle brain tissue of rats: compared with the blank group, the expression of SP and NK-1m RNA in the model group, the electroacupuncture group and the pseudo electroacupuncture group increased significantly (P0.05). Compared with the model group, the expression of SP and NK-1m RNA in the electroacupuncture group, the pseudo-needle group and the NK-1m RNA was obviously reduced (P0.05); compared with the pseudo electroacupuncture group, The expression of SP and NK-1 m RNA decreased significantly in the electroacupuncture group (P0.05). It was suggested that electroacupuncture could significantly reduce the gene expression level of SP and NK-1 in the PAG region of acute migraine, and the effect was obviously better than that of the pseudo-electroacupuncture in the treatment of.3. The number of SP and NK-1 protein in the periaqueductal gray area of the rat middle cerebral aqueduct affected the number of.SP positive cells and the blank group. Compared with the model group (group B), the number of positive cells in the pseudo electroacupuncture group (group D) increased significantly (P0.05), and there was no significant change in the electroacupuncture group (group C) (P0.05). Compared with the model group (group B), the number of positive cells in the electroacupuncture group (group C) and the pseudo electroacupuncture group (D group) decreased significantly (P0.05), and the number of positive cells in the electroacupuncture group (C group) was significantly decreased (P0.05) compared with the pseudo electroacupuncture group (D group). The expression of NK-1 positive cells: compared with the blank group (group A), the model group (group B), the electroacupuncture group (group C), the pseudo electroacupuncture group (D group) were all significantly increased, and the difference was statistically significant (P0.05). There was no significant difference between the three groups in the model group (B group), the electroacupuncture group (C group) and the pseudo electroacupuncture group (D group). It suggested that the electroacupuncture could significantly reduce the acute attack stage head. The positive distribution and expression of SP in the midbrain of the rats were significantly better than that of the pseudo-electroacupuncture treatment, and the effect on the positive expression and distribution of NK-1 had a decreasing trend, but there was no obvious change. Conclusion electroacupuncture can play a role in the prevention of migraine by reducing the pain related neuropeptide CGRP and SP expression in the acute attack stage of migraine rats.
【學(xué)位授予單位】：首都醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R245

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