本文關(guān)鍵詞：頭穴叢刺對慢性酒精中毒大鼠學(xué)習(xí)記憶能力及海馬HIF表達(dá)的影響　出處：《黑龍江中醫(yī)藥大學(xué)》2013年博士論文　論文類型：學(xué)位論文

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【摘要】：目的：通過觀察頭穴叢刺法對慢性酒精中毒所致學(xué)習(xí)記憶障礙模型大鼠學(xué)習(xí)記憶能力、HIF-1α及其靶基因VEGF mRNA和HIF mRNA表達(dá)的影響,探討頭穴叢刺對慢性酒精中毒所致學(xué)習(xí)記憶障礙的治療效果及其作用機(jī)制,為慢性酒精中毒所致腦病的臨床治療提供一定的實驗基礎(chǔ)和理論依據(jù)。 方法：將成年雄性Wistar大鼠100只,隨機(jī)分為空白對照組和酒精灌胃組。酒精灌胃組采用酒精灌胃法制備慢性酒精中毒所致學(xué)習(xí)記憶障礙大鼠模型,灌酒6周,通過Morris水迷宮檢測大鼠的學(xué)習(xí)記憶能力。再將慢性酒精中毒所致學(xué)習(xí)記憶障礙造模成功的大鼠隨機(jī)分為模型組、多奈哌齊組和頭穴叢刺組。多奈哌齊組給予多奈哌齊0.05mg/kg灌胃,1次/日,治療4周。空白對照組以同等體積的生理鹽水灌胃,灌胃量為10ml/kg,1次/日,共灌胃4周。頭穴叢刺組采用頭穴叢刺法,取百會和百會左右旁開2mm三個穴位,留針30min,1次/日,治療4周。治療4周后,采用Morris水迷宮檢測大鼠的學(xué)習(xí)記憶能力；光鏡下觀察海馬組織病理學(xué)變化；免疫組化檢測海馬HIF-1α、VEGF陽性表達(dá)；采用實時熒光定量-PCR(real-time fluorogentic quantitative PCR, RFQ-PCR)法檢測海馬HIF-1αmRNA、VEGFmRNA表達(dá)。 結(jié)果： 1.Morris水迷宮檢測結(jié)果：造模6周后,灌酒模型組大鼠學(xué)習(xí)記憶能力明顯下降,與空白對照組相比差異顯著(P0.05)；治療4周后,多奈哌齊組和頭穴叢刺組大鼠的學(xué)習(xí)記憶能力與模型組相比有所提高,具有顯著性差異(P0.01)；多奈哌齊組和頭穴叢刺組相比無顯著性差異(P0.05)。 2.HE染色結(jié)果：空白對照組未見明顯病理學(xué)變化,模型組可見大量神經(jīng)元細(xì)胞固縮,胞體形態(tài)不規(guī)則,胞漿濃縮深染,核仁變小或消失,細(xì)胞尥周圍可見明顯的空殼區(qū)；多奈哌齊組和頭穴叢剌組神經(jīng)細(xì)胞排列層次及細(xì)胞形態(tài)較清晰,少見固縮細(xì)胞,與模型組相比腑組織損害程度明顯減輕；而多奈哌齊組與頭穴叢刺組相比,固縮細(xì)胞數(shù)量明顯多于頭穴叢刺組。 3.HIF-1α、VEGF免疫組化染色結(jié)果：正常對照組神經(jīng)細(xì)胞數(shù)量、形態(tài)分布正常,排列緊密有序,細(xì)胞核圓而大,染色淺,核仁清晰,可見少量HIF-1α、VEGF陽性細(xì)胞表達(dá)；模型組、多奈哌齊組和頭穴叢刺組均可見HIF-1α、VEGF陽性細(xì)胞增加,與正常對照組比較差異具有統(tǒng)計學(xué)意義(P0.05,P0.01)；而多奈哌齊組和頭穴叢刺組較模型組陽性細(xì)胞表達(dá)減少,差異具有統(tǒng)計學(xué)意義(P0.05,P0.01)；多奈哌齊組與頭穴叢刺組相比,后者減少更為明顯,差異具有統(tǒng)計學(xué)意義(P0.01)。 4.HIFmRNA、VEGF mRNA real-time RT-PCR檢測檢測結(jié)果：空白對照組HIFmRNA、VEGF mRNA表達(dá)極弱；在模型組、多奈哌齊組和頭穴叢刺組中,HIFmRNA、VEGF mRNA均呈顯著強(qiáng)表達(dá),高于空白對照組；而多奈哌齊組和頭穴叢刺組的HIFmRNA、VEGF mRNA表達(dá)較模型組弱；頭穴叢刺組與多奈哌齊組相比,后者表達(dá)更高。 結(jié)論： 1.采用酒精灌胃法制作慢性酒精中毒所致學(xué)習(xí)記憶障礙大鼠模型,并利用Morris水迷宮從行為學(xué)和HE染色從組織學(xué)兩方面對其進(jìn)行評價,模型成功、可靠。 2.頭穴叢刺能夠改善慢性酒精中毒所致學(xué)習(xí)記憶障礙大鼠的學(xué)習(xí)記憶障礙,改善海馬組織病理學(xué)變化,能夠抑制神經(jīng)元凋亡,減少神經(jīng)細(xì)胞病理變化,從而改善學(xué)習(xí)記憶能力。 3.頭穴叢刺能夠降低HIF-1α和VEGF的陽性細(xì)胞表達(dá),減少腦損傷后的腦組織神經(jīng)元損傷,減少神經(jīng)元凋亡,發(fā)揮腦保護(hù)功能。 4.頭穴叢刺可應(yīng)用于慢性酒精中毒所致學(xué)習(xí)記憶障礙的防治,有望成為慢性酒精中毒所致學(xué)習(xí)記憶障礙新的治療方法之一。
[Abstract]:Objective: to chronic alcoholism induced learning and memory impairment of learning and memory ability of rats by observing the CASP, affecting the expression of alpha and its target gene VEGF mRNA and HIF mRNA HIF-1, to investigate the scalp acupuncture on chronic alcoholism induced learning and memory impairment in the therapeutic effect and its mechanism, provide theoretical and experimental basis on the basis of some clinical treatment for chronic alcoholism induced encephalopathy.
Methods: 100 male Wistar rats, were randomly divided into control group and alcohol gavage group. Intragastric group with alcohol by chronic alcoholism induced learning and memory impairment in rats model of gastric lavage method, alcohol for 6 weeks, the learning and memory of rats in Morris water maze test can then. Chronic alcoholism induced impairment of learning and memory of rats were randomly divided into model group, donepezil group and scalp acupuncture group. Donepezil group were given donepezil 0.05mg/kg orally, 1 times / day, for 4 weeks. Blank control group with the same volume of saline by gavage, gastric lavage volume is 10ml/kg, 1 time / day, a total of 4 weeks. Head acupuncture group with head acupuncture method, Baihui and Baihui about 2mm beside the three points for 30min, 1 times / day, for 4 weeks. After 4 weeks of treatment, the ability of learning and memory in Morris water maze test in rats were observed; in the light microscope Histopathological changes, immunohistochemistry, HIF-1 positive expression of VEGF and -PCR in hippocampus were detected, and HIF-1 mRNA mRNA and VEGFmRNA expression in hippocampus were detected by real-time fluorogentic quantitative PCR (RFQ-PCR).
Result錛，

本文編號：1420551