本文選題：失眠 + 針刺　； 參考：《成都中醫(yī)藥大學(xué)》2016年碩士論文

【摘要】：目的：借助PCPA失眠動(dòng)物模型,觀察正常和失眠SD大鼠自發(fā)活動(dòng)晝夜節(jié)律特征,以及針刺蹺脈后自發(fā)活動(dòng)晝夜節(jié)律的調(diào)整和變化；觀察失眠大鼠海馬內(nèi)5-HT1A、5-HT2A受體mRNA及蛋白表達(dá)的改變,評(píng)價(jià)針刺蹺脈對(duì)失眠大鼠的治療作用,以探討其內(nèi)在作用機(jī)制,為針刺蹺脈治療失眠提供科學(xué)依據(jù)。方法：1.將30只雄性SD大鼠馴化與L：D(12：12)光暗同步后,利用動(dòng)物自發(fā)活動(dòng)自動(dòng)記錄及節(jié)律分析系統(tǒng)和美國CLOCKLAB數(shù)據(jù)采集分析系統(tǒng),自動(dòng)檢測(cè)和記錄自發(fā)活動(dòng)晝夜節(jié)律。然后將SD大鼠隨機(jī)分為空白組、模型組、針刺組,每組10只。2.對(duì)模型組和針刺組SD大鼠,運(yùn)用腹腔注射對(duì)氯苯丙氨酸(PCPA)混懸液的方法,復(fù)制失眠模型。觀察造模前后自發(fā)活動(dòng)晝夜節(jié)律的改變。3.造模后,每日早上8：00-10：00,對(duì)空白組及模型組給予抓放固定、不針刺干預(yù),對(duì)針刺組給予抓放固定、針刺“申脈”、“照�！备深A(yù),1次／天,20min/次,連續(xù)1周,觀察針刺蹺脈對(duì)自發(fā)活動(dòng)晝夜節(jié)律的影響。4.應(yīng)用實(shí)時(shí)熒光定量PCR(RT-PCR)檢測(cè)海馬內(nèi)5-HT1A、5-HT2A受體mRNA的表達(dá)水平。5.應(yīng)用蛋白免疫印跡(Western Blot)檢測(cè)海馬內(nèi)5-HT1A、5-HT2A受體蛋白的表達(dá)水平。結(jié)果：1.在L：D(12：12)馴化穩(wěn)定后,SD大鼠自發(fā)活動(dòng)均表現(xiàn)出近似晝夜節(jié)律。模型組與針刺組在PCPA造模后,表現(xiàn)出明顯的自發(fā)活動(dòng)晝夜節(jié)律紊亂,休息期活動(dòng)量明顯增加,與空白組比較有統(tǒng)計(jì)學(xué)意義(P0.05)。2.針刺蹺脈對(duì)針刺組大鼠自發(fā)活動(dòng)晝夜節(jié)律具有調(diào)整作用,休息期活動(dòng)量減少,與模型組比較,有統(tǒng)計(jì)學(xué)意義(P0.05)。3.造模后,模型組大鼠海馬中5-HT1A受體mRNA相對(duì)表達(dá)值減少,5-HT2A受體mRNA相對(duì)表達(dá)值增加,與空白組比較,有統(tǒng)計(jì)學(xué)意義(P0.05)。4.針刺治療后,針刺組大鼠海馬中5-HT1A受體mRNA相對(duì)表達(dá)值增加,5-HT2A受體mRNA相對(duì)表達(dá)值減少,與模型組比較,有統(tǒng)計(jì)學(xué)意義(P0.05)。5.造模后,模型組大鼠海馬中5-HT1A受體蛋白的表達(dá)水平減少,5-HT2A受體蛋白的表達(dá)水平增加,與空白組比較,有統(tǒng)計(jì)學(xué)意義(P0.05)。6.針刺治療后,針刺組大鼠海馬中5-HT1 A受體蛋白的表達(dá)水平增加,5-HT2A受體蛋白的表達(dá)水平減少,與模型組比較,有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論：1.PCPA造模后失眠SD大鼠一般狀態(tài)表現(xiàn)異常,自發(fā)活動(dòng)晝夜節(jié)律紊亂,休息期活動(dòng)量明顯增加。2.針刺蹺脈可以使失眠SD大鼠自發(fā)活動(dòng)晝夜節(jié)律恢復(fù)其規(guī)律性,休息期活動(dòng)量相對(duì)減少,睡眠逐漸恢復(fù)正常。3.針刺蹺脈可以使失眠SD大鼠海馬中5-HT1A受體mRNA及蛋白表達(dá)增加,5-HT2A受體mRNA及蛋白表達(dá)減少。這可能是針刺蹺脈治療失眠的內(nèi)在作用機(jī)制之一。
[Abstract]:Objective: to observe the circadian rhythm of spontaneous activity in normal and insomnia SD rats, and to observe the regulation and change of the circadian rhythm of spontaneous activity after acupuncture.To observe the changes of mRNA and protein expression of 5-HT _ (1) A _ (5-HT _ 2A) receptor in hippocampus of insomnia rats, to evaluate the therapeutic effect of acupuncture on stilt veins in insomnia rats, to explore its intrinsic mechanism, and to provide scientific basis for acupuncture of stilt-vein acupuncture in the treatment of insomnia.Method 1: 1.30 male Sprague-Dawley rats were acclimated to L: D12: 12) light and dark synchronization. The spontaneous activity circadian rhythm was automatically detected and recorded by the automatic recording and rhythm analysis system of animal spontaneous activity and the CLOCKLAB data acquisition and analysis system of the United States.Then SD rats were randomly divided into blank group, model group, acupuncture group, 10 rats in each group.Models of insomnia were established in SD rats in model group and acupuncture group by intraperitoneal injection of PCPA suspension.Observe the changes of circadian rhythm of spontaneous activity before and after modeling.After making the model, 8: 00-10: 00 every morning, the blank group and the model group were given grip fixation without acupuncture intervention, and the acupuncture group was given grasping fixation, acupuncture "Shenmai" and "Zhaohai" intervention once a day for 20 minutes every time for one week in a row.To observe the effect of acupuncture on the circadian rhythm of spontaneous activity.The expression level of 5-HT _ 1A _ (5-HT _ 2A) receptor mRNA in hippocampus was detected by real-time quantitative PCR- RT-PCR- PCR.Western blot was used to detect the expression of 5-HT _ 1A _ (5-HT _ 2A) receptor protein in hippocampus.The result is 1: 1.The spontaneous activity of SD rats showed an approximate circadian rhythm after acclimation and stabilization.After PCPA, the model group and acupuncture group showed obvious disturbance of the circadian rhythm of spontaneous activity and increased the amount of activity during rest period, which had statistical significance compared with the blank group (P0.05. 2).Acupuncture of still-vein can adjust the circadian rhythm of spontaneous activity of rats in acupuncture group and decrease the amount of activity in rest period. Compared with the model group, it has statistical significance (P0.05 路3).After modeling, the relative expression of 5-HT1A receptor mRNA in the hippocampus of the model group decreased and the relative expression value of 5-HT2A receptor mRNA increased. Compared with the blank group, the expression of 5-HT1A receptor mRNA in the model group was significantly higher than that in the blank group.After acupuncture treatment, the relative expression of 5-HT1A receptor mRNA in hippocampus of the acupuncture group increased and the relative expression value of 5-HT2A receptor mRNA decreased. Compared with the model group, the relative expression of 5-HT1A receptor mRNA was significantly higher than that of the model group.After modeling, the expression of 5-HT1A receptor protein in the hippocampus of the model group decreased and the expression level of 5-HT2A receptor protein increased. Compared with the blank group, the expression level of 5-HT1A receptor protein in the model group was significantly higher than that in the blank group.After acupuncture treatment, the expression level of 5-HT1 A receptor protein in hippocampus of the acupuncture group increased and the expression level of 5-HT2A receptor protein decreased. Compared with the model group, the expression level of 5-HT1 A receptor protein was significantly higher than that of the model group (P 0.05).Conclusion 1. The normal state of insomnia SD rats was abnormal, the circadian rhythm of spontaneous activity was disordered, and the amount of activity during rest period was increased significantly.Acupuncture can restore the circadian rhythm of spontaneous activity of insomnia SD rats, decrease the amount of activity during rest period, and gradually return to normal. 3.Acupuncture could increase the expression of 5-HT1A receptor mRNA and protein in hippocampus of SD rats and decrease the expression of 5-HT 2A receptor mRNA and protein.This may be one of the internal mechanisms of acupuncture on the treatment of insomnia.
【學(xué)位授予單位】：成都中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R245

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