本文選題：電針 + 腦缺血再灌注�。� 參考：《南京中醫(yī)藥大學》2016年碩士論文

【摘要】：目的：通過觀察電針對腦缺血再灌注大鼠模型細胞凋亡相關基因Bcl-2、Bax蛋白表達的影響,探討電針改善腦缺血后細胞凋亡的調(diào)節(jié)作用,以期為針刺治療缺血性腦中風的效應機制的闡釋提供科學依據(jù)。方法：選用SD大鼠按照隨機數(shù)字表抽取24只為假手術組,余下動物采用改良Longa線栓法建立腦缺血再灌注模型。將造模成功的大鼠進行隨機分組,分別為模型組、電針組,每組有效例數(shù)為24只。假手術組除了不穿入線栓外,其他處理與模型組相同；模型組制備模型缺血2h后拔出線栓,不進行任何治療處理；電針組用一次性針灸針選取“大椎”、“百會”穴進行刺激,“百會”穴平刺,“大椎”穴約30。斜刺,深度約1cm,接韓氏HANS-200型電針儀,“百會”穴接負極,“大椎”穴接正極,選擇疏密波,頻率為2/15Hz,刺激強度以針柄輕微顫動,大鼠不發(fā)出叫聲為度,連續(xù)治療30min。腦缺血再灌注24h后,參照Julio氏18分制評分法進行神經(jīng)功能行為學評分；采用干濕重稱量法計算腦含水量,腦組織含水量(%)=(濕重一干重)/濕重×100%；采用免疫組化法和Rt-PCR技術分別檢測缺損側大腦皮層Bcl-2. Bax蛋白表達及Bcl-2mRNA、Bax mRNA的表達變化。結果：(1)腦缺血再灌注損傷大鼠神經(jīng)功能評分明顯降低,與假手術組比較差異顯著(P0.01)；電針干預治療能有效提高其評分值,與模型組比較有顯著差異(P0.05)。(2)腦缺血再灌注損傷大鼠腦含水量明顯高于假手術組(P0.01),與模型組比較電針干預后腦含水量顯著降低(P0.01),并接近假手術組(P0.05)。(3)缺損側大腦皮層Bcl-2、Bax蛋白的表達,模型組均較假手術組明顯升高(P0.05和P0.01),其中Bax蛋白的升高幅度大于Bcl-2蛋白,Bcl-2/Bax蛋白的比值減小(P0.01)；電針組與模型組比較,Bcl-2蛋白的表達升高(P0.05), Bax蛋白的表達降低(P0.01), Bcl-2/Bax蛋白的比值升高(P0.01)。(4)缺損側大腦皮層Bcl-2mRNA、BaxmRNA的表達,模型組均較假手術組明顯升高(P0.05),Bcl-2mRNA/Bax mRNA的比值降低(P0.05)；與模型組比較,電針組Bcl-2mRNA的升高不明顯(P0.05),但Bax mRNA的降低顯著(P0.05),Bcl-2mRNA/BaxmRNA的比值升高(P0.01)。結論：電針對缺血再灌注腦損傷大鼠Bcl-2、Bax基因具有調(diào)控作用,提升Bcl-2/Bax的比值,使抗凋亡基因占據(jù)優(yōu)勢,降低缺血再灌注區(qū)的細胞凋亡,減輕腦水腫,改善神經(jīng)功能。
[Abstract]:Objective: to observe the effect of electroacupuncture on the expression of apoptosis-related gene Bcl-2nBax in rat model of cerebral ischemia-reperfusion, and to explore the regulatory effect of electroacupuncture on apoptosis after cerebral ischemia. In order to provide scientific basis for the explanation of the effect mechanism of acupuncture on ischemic stroke. Methods: 24 SD rats were selected as sham-operated group according to random number table. The model of cerebral ischemia-reperfusion was established by modified Longa method. The successful rats were randomly divided into model group and electroacupuncture group with 24 effective cases in each group. The sham-operation group had the same treatment as the model group except for not wearing the thread embolus; the model group prepared the model with 2 hours of ischemia and pulled out the thread plug without any treatment; the electroacupuncture group used a one-off acupuncture needle to select "Dazhui" and "Baihui" to stimulate. "Baihui" point flat thorn, "Dazhui" point about 30. Oblique prickles, depth about 1 cm, HANS-200 type electroacupuncture apparatus, "Baihui" point and "Dazhui" point were connected with negative electrode, and "Dazhui" point was connected with positive pole. The frequency of dense wave was 2 / 15 Hz. The stimulation intensity was slightly quivered with needle handle, and the rats did not emit a call for 30 mins. After 24 hours of cerebral ischemia-reperfusion, the neurobehavioral score was evaluated with Julio's 18 score system, and the water content of brain was calculated by using dry and wet weighing method. The contents of water in brain tissue were as follows: (wet weight, dry weight, wet weight 脳 100), immunohistochemical method and Rt-PCR technique were used to detect Bcl-2 in the damaged cerebral cortex. Expression of Bax protein and Bcl-2 mRNA-Bax mRNA. Results the neurological function score of rats with cerebral ischemia-reperfusion injury was significantly lower than that of sham operation group (P 0.01). Compared with the model group, the brain water content of rats with cerebral ischemia-reperfusion injury was significantly higher than that of the sham-operated group, and the brain water content of the model group was significantly lower than that of the model group after electroacupuncture intervention, and close to the expression of Bcl-2Bax protein in the cortical cortex of the sham-operated group. The expression of Bcl-2 / Bax protein in the model group was significantly higher than that in the sham operation group (P 0.05 and P 0.01), in which the increase of Bax protein was greater than that in the Bcl-2 protein Bcl-2 / Bax protein ratio. The expression of Bcl-2 protein in the electroacupuncture group was significantly higher than that in the model group, and the expression of Bax protein was lower than that in the model group (P 0.01, Bcl-2/Bax protein). The expression of Bcl-2mRNA-mRNA-Bax mRNA in the cortical cortex of the defect side was increased with the increase of the ratio of Bcl-2mRNA-mRNA-BaxmRNA. Compared with the model group, the ratio of P0.05mRNA-2mRNA-Bax mRNA in the model group was significantly higher than that in the sham-operated group, but the ratio of P0.05mRNA-2mRNA-Bax mRNA in the electroacupuncture group was not significantly increased than that in the model group, but the decrease of Bax mRNA was significantly higher than that of P0.05mRNA-2mRNA-Bax mRNA. Conclusion: electroacupuncture can regulate the Bcl-2P Bax gene in rats with cerebral ischemia-reperfusion injury, increase the ratio of Bcl-2/Bax, make anti-apoptotic genes dominant, reduce apoptosis in ischemia-reperfusion area, reduce brain edema and improve neural function.
【學位授予單位】：南京中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R245

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