【摘要】：目的通過水迷宮實驗,觀察鹽酸美金剛(memantine hydrochloride)對血管性癡呆(vascular dementia,Va D)大鼠學習記憶能力的影響;通過對大鼠海馬CA3-CA1區(qū)長時程增強(long-term potentiation,LTP)及N-甲基-D天冬氨酸受體(N-methy-Daspartate receptor,NMDAR)各亞基和腦源性神經(jīng)營養(yǎng)因子(brain-derived neurotrophic factor,BDNF)表達的測定,分析鹽酸美金剛改善Va D大鼠學習記憶能力的作用機制。方法將30只成年雄性SD大鼠隨機分為假手術組、Va D模型組和美金剛組,每組10只。Va D模型組和美金剛組采用永久性結扎雙側頸總動脈方法制作Va D大鼠模型。美金剛組于術后1周給予美金剛[10mg/(kg·d)]灌胃治療,Va D模型組給予等量生理鹽水,共干預4周。于術后5周采用Morris水迷宮測試各組大鼠的學習記憶水平,包括定位航行實驗和空間探索實驗,計算平均逃避潛伏期及目標象限百分率。各組大鼠隨機抽取6只行LTP檢測各組大鼠海馬區(qū)的突觸可塑性,后斷頭取腦,用Western Blot方法測定各組大鼠海馬區(qū)BDNF及NMDAR各亞基(NR1、NR2A、NR2B、NR2C、NR2D)蛋白的表達水平;各組剩余大鼠于水迷宮檢測后斷頭取腦,用HE染色方法觀察各組大鼠海馬區(qū)細胞形態(tài)的變化;應用免疫組織化學方法檢測各組大鼠海馬區(qū)BDNF、NR1的表達。結果1、Morris水迷宮測試結果與假手術組比較,Va D模型組大鼠水迷宮實驗逃避潛伏期顯著延長(P0.05),目標象限百分率明顯下降(P0.05)。與模型組比較,美金剛組大鼠水迷宮實驗逃避潛伏期顯著下降(P0.05),目標象限百分率明顯上升(P0.05)。2、LTP檢測結果Va D模型組大鼠海馬CA3-CA1區(qū)LTP受損明顯(P0.05),美金剛組大鼠海馬CA3-CA1區(qū)LTP明顯優(yōu)于模型組(P0.05)。3、Western Blot結果與假手術組比較,Va D模型組大鼠海馬區(qū)BDNF、NR1、NR2A、NR2B蛋白表達水平明顯下降(P0.05),NR2D蛋白表達增加(P0.05)。與模型組比較,美金剛組大鼠BDNF、NR1、NR2A、NR2B蛋白的表達顯著增加(P0.05),但仍低于假手術組(P0.05);NR2D蛋白表達下降(P0.05),但仍高于假手術組(P0.05)。各組大鼠海馬區(qū)NR2C蛋白表達變化無統(tǒng)計學意義(P0.05)4、HE染色與假手術組比較,Va D模型組大鼠海馬區(qū)神經(jīng)細胞數(shù)量明顯減少,排列紊亂,細胞大量萎縮,細胞形態(tài)破壞,出現(xiàn)空泡變性、核固縮,細胞器結構不清。與模型組相比,美金剛組神經(jīng)細胞數(shù)量增多,排列較整齊,細胞形態(tài)較完整,僅見少量核固縮。5、免疫組化染色結果Va D模型組大鼠海馬CA1區(qū)錐體細胞BDNF、NR1表達量較假手術組明顯減少(P0.05),美金剛組海馬CA1區(qū)錐體細胞BDNF、NR1表達量比模型組顯著增多(P0.05)。結論1、永久性結扎雙側頸總動脈制作的Va D模型可以模擬人類血管性癡呆的病理機制。2、美金剛可改善Va D大鼠的空間學習記憶能力及突觸可塑性。3、美金剛可通過上調(diào)BDNF、NR1、NR2A和NR2B的表達,下調(diào)NR2D的表達,調(diào)節(jié)突觸可塑性,增強LTP,改善Va D大鼠的學習記憶水平。
[Abstract]:Objective to observe the effect of (memantine hydrochloride) on the learning and memory ability of vascular dementia (vascular dementia,Va D) rats by water maze test. The expression of long term potentiation (long-term potentiation,LTP), N-methyl-D-aspartate receptor (N-methy-Daspartate receptor,NMDAR) subunits and brain-derived neurotrophic factor (brain-derived neurotrophic factor,BDNF) in rat hippocampal CA3-CA1 were measured. Objective: to analyze the mechanism of improving learning and memory ability of Va D rats by methadine hydrochloride. Methods Thirty adult male SD rats were randomly divided into sham-operation group (, Va D model group) and meringang group (10 rats in each group). Va D rats were made by permanent ligation of bilateral common carotid artery in each group. The, Va D model group was treated with 10mg/ (kg d) for 4 weeks. The learning and memory levels of each group were measured by Morris water maze at 5 weeks after operation, including navigation experiment and space exploration experiment. The average escape latency and the percentage of target quadrant were calculated. Six rats in each group were randomly selected for LTP to detect the synaptic plasticity in the hippocampal area of each group, and then the brain was taken from the head off. The expression levels of BDNF and NMDAR subunits (NR1,NR2A,NR2B,NR2C,NR2D) in the hippocampal area of each group were measured by Western Blot method. The remaining rats in each group were taken out of their heads after water maze detection, and the changes of cell morphology in hippocampus were observed by HE staining, and the expression of BDNF,NR1 in hippocampus was detected by immunohistochemical method. Results 1 compared with sham operation group, the escape latency of water maze test in, Va D group was significantly prolonged (P0.05), and the percentage of target quadrant was significantly decreased (P0.05). Compared with the model group, the escape latency of the water maze test in the MJ group decreased significantly (P0.05), and the percentage of the target quadrant increased significantly (P0.05). The results of LTP test showed that the LTP of CA3-CA1 in hippocampus of Va D group was significantly damaged (P0.05), and the LTP of CA3-CA1 area of hippocampus in MJ group was significantly better than that in model group (P0.05). 3 the results of Western Blot were compared with those of sham-operation group. In Va D group, the expression of BDNF,NR1,NR2A,NR2B protein decreased significantly (P0.05), and the expression of NR2D protein increased (P0.05). Compared with the model group, the expression of BDNF,NR1,NR2A,NR2B protein was significantly increased (P0.05), but still lower than that of sham-operated group (P0.05); the expression of NR2D protein decreased (P0.05), but still higher than that of sham-operated group (P0.05). There was no significant change of NR2C protein expression in hippocampus of rats in each group (P0.05). (4) compared with sham-operation group, the number of hippocampal neurons in, Va D model group was significantly decreased, the number of neurons in hippocampus was disordered, and the number of cells was atrophy. Cell morphological destruction, vacuolar degeneration, nuclear pyknosis, unclear organelle structure. Compared with the model group, the number of neurons in the MJ group increased, arranged neatly, the morphology of the cells was relatively complete, and only a small number of nuclear pyknosis was observed. The results of immunohistochemical staining showed that the pyramidal cells in the hippocampal CA1 region of the rats in the Va D model group were BDNF,. The expression of NR1 was significantly lower than that of sham-operated group (P0.05), and the expression of BDNF,NR1 in hippocampal CA1 pyramidal cells in MJ group was significantly higher than that in model group (P0.05). Conclusion 1. The Va D model made by permanent ligation of bilateral common carotid arteries can mimic the pathological mechanism of human vascular dementia. (2) MJ can improve the spatial learning and memory ability and synaptic plasticity of Va D rats. By upregulating the expression of BDNF,NR1,NR2A and NR2B, decreasing the expression of NR2D, regulating synaptic plasticity, and enhancing LTP, to improve the learning and memory of Va D rats, MJ could improve the learning and memory level of Va D rats.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R749.13

【共引文獻】

相關期刊論文 前10條

1 王巖峰;秦光華;張玉強;楊明超;王U，

本文編號：2313077