盐酸美金刚对血管性痴呆大鼠学习记忆障碍的影响及作用机制研究
发布时间：2018-11-05 19:04
【摘要】：目的通过水迷宫实验,观察盐酸美金刚(memantine hydrochloride)对血管性痴呆(vascular dementia,Va D)大鼠学习记忆能力的影响;通过对大鼠海马CA3-CA1区长时程增强(long-term potentiation,LTP)及N-甲基-D天冬氨酸受体(N-methy-Daspartate receptor,NMDAR)各亚基和脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)表达的测定,分析盐酸美金刚改善Va D大鼠学习记忆能力的作用机制。方法将30只成年雄性SD大鼠随机分为假手术组、Va D模型组和美金刚组,每组10只。Va D模型组和美金刚组采用永久性结扎双侧颈总动脉方法制作Va D大鼠模型。美金刚组于术后1周给予美金刚[10mg/(kg·d)]灌胃治疗,Va D模型组给予等量生理盐水,共干预4周。于术后5周采用Morris水迷宫测试各组大鼠的学习记忆水平,包括定位航行实验和空间探索实验,计算平均逃避潜伏期及目标象限百分率。各组大鼠随机抽取6只行LTP检测各组大鼠海马区的突触可塑性,后断头取脑,用Western Blot方法测定各组大鼠海马区BDNF及NMDAR各亚基(NR1、NR2A、NR2B、NR2C、NR2D)蛋白的表达水平;各组剩余大鼠于水迷宫检测后断头取脑,用HE染色方法观察各组大鼠海马区细胞形态的变化;应用免疫组织化学方法检测各组大鼠海马区BDNF、NR1的表达。结果1、Morris水迷宫测试结果与假手术组比较,Va D模型组大鼠水迷宫实验逃避潜伏期显著延长(P0.05),目标象限百分率明显下降(P0.05)。与模型组比较,美金刚组大鼠水迷宫实验逃避潜伏期显著下降(P0.05),目标象限百分率明显上升(P0.05)。2、LTP检测结果Va D模型组大鼠海马CA3-CA1区LTP受损明显(P0.05),美金刚组大鼠海马CA3-CA1区LTP明显优于模型组(P0.05)。3、Western Blot结果与假手术组比较,Va D模型组大鼠海马区BDNF、NR1、NR2A、NR2B蛋白表达水平明显下降(P0.05),NR2D蛋白表达增加(P0.05)。与模型组比较,美金刚组大鼠BDNF、NR1、NR2A、NR2B蛋白的表达显著增加(P0.05),但仍低于假手术组(P0.05);NR2D蛋白表达下降(P0.05),但仍高于假手术组(P0.05)。各组大鼠海马区NR2C蛋白表达变化无统计学意义(P0.05)4、HE染色与假手术组比较,Va D模型组大鼠海马区神经细胞数量明显减少,排列紊乱,细胞大量萎缩,细胞形态破坏,出现空泡变性、核固缩,细胞器结构不清。与模型组相比,美金刚组神经细胞数量增多,排列较整齐,细胞形态较完整,仅见少量核固缩。5、免疫组化染色结果Va D模型组大鼠海马CA1区锥体细胞BDNF、NR1表达量较假手术组明显减少(P0.05),美金刚组海马CA1区锥体细胞BDNF、NR1表达量比模型组显著增多(P0.05)。结论1、永久性结扎双侧颈总动脉制作的Va D模型可以模拟人类血管性痴呆的病理机制。2、美金刚可改善Va D大鼠的空间学习记忆能力及突触可塑性。3、美金刚可通过上调BDNF、NR1、NR2A和NR2B的表达,下调NR2D的表达,调节突触可塑性,增强LTP,改善Va D大鼠的学习记忆水平。
[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.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R749.13
[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.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R749.13
【共引文献】
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1 王岩峰;秦光华;张玉强;杨明超;王U，
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