维生素E干预对锰中毒小鼠学习记忆及神经干细胞的影响
发布时间：2019-05-27 09:23
【摘要】：目的:采用神经行为学 组织化学等方法研究锰中毒对小鼠海马区神经细胞的影响生素E对其干预作用,并探讨其发生机制,为锰中毒的机理 预治疗提供实验依据。方法:1、动物 组:选用清洁级成年雄性健康的昆明小鼠75只,体重在18-22g(广西医科大学实验动物中心提供)。将选用的75只昆明小鼠按随机对照方法分为5组,分别是对照组(CG)、生素E组(VG)、锰中毒组(MG)、锰加生素E组(MVG)、锰加普罗布考组(MPG),每组15只。2、制作锰中毒小鼠模型:分别对锰中毒、锰加生素E组、锰加普罗布考组3组小鼠腹腔注射氯化锰(Mncl2),剂量为20mg/kg,其余2组小鼠腹腔注射等容积的生理盐水。每只小鼠注射的量为0.01ml/g,每天下午注射1次,连续注射4周。3、药物灌胃:分别对生素E组、锰加生素E组行生素E药物灌胃干预,剂量为100mg/kg,对照组和锰中毒组行等容积的橄榄油灌胃,锰加普罗布考组予以普罗布考药物灌胃,剂量为500mg/kg,每只小鼠灌胃的量是0.01ml/g,各组小鼠每天下午灌胃1次,连续给药4周。各组小鼠在相同环境下饲养,自由饮食。4、神经行为学检测:4周后用Morris水迷宫检测5组小鼠的逃避潜伏期和穿越平台次数,从而测定各组小鼠的学习记忆能力。5、免疫组织化学染色:用免疫组化的方法分别检测每组小鼠海马CA1区和DG区caspase-3、细胞色素C的表达和海马DG区nestin的表达。计算所得免疫组织化学图像的阳性细胞数,用统计软件spss16.0和Graph Pad Prism5统计分析各组小鼠Morris水迷宫所得数据和免疫组化结果。结果:1、Morris水迷宫检测结果:水迷宫检测实验有定位航行实验和空间探索实验两部分组成,定位航行实验中,与CG相比,MG小鼠的逃避潜伏期明显延长,差异有统计学意义(P0.05);与MG相比,MVG和MPG小鼠的逃避潜伏期明显缩短,差异有统计学意义(P0.05);MVG和MPG小鼠的逃避潜伏期相比较,差异无统计学意义(P0.05);CG和VG小鼠的逃避潜伏期相比较,差异无统计学意义(P0.05)。在空间探索实验中,与CG相比,MG小鼠的穿越平台的次数明显减少,差异有统计学意义(P0.05);而与MG比较,MVG和MPG小鼠穿越平台次数明显增多,差异有统计学意义(P0.05);MVG和MPG小鼠穿越平台次数相比无明显差别,差异无统计学意义(P0.05);CG和VG小鼠的穿越平台次数无明显差别,差异无统计学意义(P0.05)。2、免疫组织化学细胞色素C的表达:与CG相比,MG海马CA1区和D G区细胞色素C的表达明显增多,差异有统计学意义(P0.05);与MG相比,MVG和MPG小鼠细胞色素C的表达明显减少,有统计学差异(P0.05);CG和VG之间海马CA1区和DG区细胞色素C表达的比较差异无统计学意义(P0.05);同时MVG和MPG间比较差异无统计学意义(P0.05)。3、免疫组织化学caspase-3的表达:与CG相比较,MG海马CA1区和DG区caspase-3阳性细胞数明显增多,差异有统计学意义(P0.05);与MG相比,MVG和MPG小鼠caspase-3的表达减少,差异有显著性(P0.05);而CG和VG之间比较差异无统计学意义(P0.05);同时MVG和MPG间比较差异无统计学意义(P0.05)。4、免疫组织化学nestin的表达:与CG相比较,MG海马齿状回nestin的阳性细胞数明显减少,差异有统计学意义(P0.05);NVG与MG相比,其nestin的表达明显增多;而MG和MPG间nestin的表达无明显差别(P0.05);同时CG和VG间比较差异无统计学意义(P0.05)。结论:1、生素E和和普罗布考干预均可改善锰中毒小鼠的学习记忆能力;2.锰中毒可促进小鼠脑内细胞色素C的增多,上调caspase-3的表达,从而降低小鼠的学习记忆能力;3、生素E和普罗布考可减少锰中毒小鼠细胞色素C和caspase-3的表达,减少神经细胞的凋亡;4、锰中毒可减少脑内DG区神经干细胞,生素E可增加锰中毒小鼠脑内神经干细胞的数量,而普罗布考对锰中毒小鼠脑内神经干细胞没有影响;5、生素E可能主要通过抗氧化作用,来对改善锰中毒小鼠的学习记忆能力,同时生素E通过增加锰中毒脑内DG区神经干细胞数量,对锰中毒脑损伤的修复有一定的保护作用。
[Abstract]:Objective: To study the effect of manganese poisoning on the nerve cells in the hippocampus of mice by using the method of neurobehavioral histochemistry and the like, and to explore the mechanism of its occurrence and provide the experimental basis for the mechanism of manganese poisoning. Methods:1, animal group:75 male healthy Kunming mice were selected and the body weight was 18-22 g (provided by the experimental animal center of Guangxi Medical University). The selected 75 Kunming mice were divided into 5 groups according to the randomized controlled method. The control group (CG), the prebiotic E group (VG), the manganese poisoning group (MG), the manganese prebiotic E group (MVG) and the manganese plus probucol group (MPG) were divided into 5 groups. Manganese poisoning, manganese prebiotic E group, manganese plus probucol group 3 mice were injected intraperitoneally with manganese chloride (MnCl2) at a dose of 20 mg/ kg, and the rest of 2 mice were injected with saline in the same volume as the abdominal cavity. the injection amount of each mouse is 0.01ml/ g, and is injected every afternoon for 1 time, and is continuously injected for 4 weeks.3, the medicine is fed into the stomach by means of the intragastric administration of the prebiotic E, the dosage of which is 100 mg/ kg, the control group and the manganese poisoning group line, and the like, The dose of probucol was 500 mg/ kg and the amount of each mouse was 0.01 ml/ g. The mice were fed in the same environment and the free diet.4. Neurobehavioral test:4 weeks after 4 weeks, Morris water maze was used to detect the escape latency of 5 mice and the number of crossing platforms, so as to determine the learning and memory abilities of each group of mice. The expression of caspase-3, the expression of cytochrome C and the expression of nestin in the hippocampus of the hippocampus of each group were detected by immunohistochemistry. The numbers of the positive cells of the obtained immunohistochemical images were calculated, and the data and the immunohistochemical results of the Morris water maze of each group were statistically analyzed by using the statistical software sps16.0 and the Graph Pad Prim5. Results:1. The result of the Morris water maze test: The water maze test was made up of two parts: the positioning navigation experiment and the space exploration experiment. Compared with the CG, the escape latency of the MG mice was significantly prolonged compared with that of the CG (P0.05). The escape latency of MVG and MPG mice was significantly shortened, and the difference was significant (P0.05). The difference of the escape latency of MVG and MPG mice was not significant (P0.05). In the experiment of space exploration, the number of crossing platforms in MG mice was significantly decreased compared with CG (P0.05). Compared with MG, the number of crossing platforms of MVG and MPG mice increased significantly (P0.05). There was no significant difference in the number of crossing platforms of MVG and MPG mice (P0.05). There was no significant difference in the number of crossing platforms in CG and VG mice (P0.05). The expression of cytochrome C in the CA1 region and the D G region of MG was significantly increased, and the difference was significant (P0.05). Compared with MG, the expression of cytochrome C in the MVG and MPG mice was significantly reduced, and there was a statistical difference (P0.05). There was no significant difference in the expression of cytochrome C in the CA1 and DG regions of the hippocampus between CG and VG (P0.05), and the difference between MVG and MPG was not significant (P0.05). There was a significant difference in the expression of caspase-3 in MVG and MPG mice (P0.05). Compared with CG, the number of positive cells in the dentate gyrus of MG was significantly decreased, and the difference was significant (P0.05); the expression of nestin in MG and MPG was significantly higher than that of MG (P0.05); and the difference between CG and VG was not statistically significant (P0.05). Conclusion:1. Prebiotic E and propafenicol can improve the learning and memory ability of manganese-poisoned mice. Manganese poisoning can promote the increase of cytochrome C in the mouse brain, increase the expression of caspase-3, and reduce the learning and memory capacity of the mouse; and 3, the prebiotic E and the probucol can reduce the expression of the cytochrome C and the caspase-3 in the manganese-poisoned mice, and reduce the apoptosis of the nerve cells; and 4, Manganese poisoning can reduce the number of neural stem cells in the brain of the brain, which can increase the number of neural stem cells in the brain of the mice, and the probucol has no effect on the neural stem cells in the brain of the mice. So as to improve the learning and memory capacity of the manganese-poisoned mice, and meanwhile, the prebiotics E has a certain protective effect on the repair of the brain damage of the manganese poisoning by increasing the number of the neural stem cells in the DG region in the brain of the manganese poisoning.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R595
本文编号：2486035
[Abstract]:Objective: To study the effect of manganese poisoning on the nerve cells in the hippocampus of mice by using the method of neurobehavioral histochemistry and the like, and to explore the mechanism of its occurrence and provide the experimental basis for the mechanism of manganese poisoning. Methods:1, animal group:75 male healthy Kunming mice were selected and the body weight was 18-22 g (provided by the experimental animal center of Guangxi Medical University). The selected 75 Kunming mice were divided into 5 groups according to the randomized controlled method. The control group (CG), the prebiotic E group (VG), the manganese poisoning group (MG), the manganese prebiotic E group (MVG) and the manganese plus probucol group (MPG) were divided into 5 groups. Manganese poisoning, manganese prebiotic E group, manganese plus probucol group 3 mice were injected intraperitoneally with manganese chloride (MnCl2) at a dose of 20 mg/ kg, and the rest of 2 mice were injected with saline in the same volume as the abdominal cavity. the injection amount of each mouse is 0.01ml/ g, and is injected every afternoon for 1 time, and is continuously injected for 4 weeks.3, the medicine is fed into the stomach by means of the intragastric administration of the prebiotic E, the dosage of which is 100 mg/ kg, the control group and the manganese poisoning group line, and the like, The dose of probucol was 500 mg/ kg and the amount of each mouse was 0.01 ml/ g. The mice were fed in the same environment and the free diet.4. Neurobehavioral test:4 weeks after 4 weeks, Morris water maze was used to detect the escape latency of 5 mice and the number of crossing platforms, so as to determine the learning and memory abilities of each group of mice. The expression of caspase-3, the expression of cytochrome C and the expression of nestin in the hippocampus of the hippocampus of each group were detected by immunohistochemistry. The numbers of the positive cells of the obtained immunohistochemical images were calculated, and the data and the immunohistochemical results of the Morris water maze of each group were statistically analyzed by using the statistical software sps16.0 and the Graph Pad Prim5. Results:1. The result of the Morris water maze test: The water maze test was made up of two parts: the positioning navigation experiment and the space exploration experiment. Compared with the CG, the escape latency of the MG mice was significantly prolonged compared with that of the CG (P0.05). The escape latency of MVG and MPG mice was significantly shortened, and the difference was significant (P0.05). The difference of the escape latency of MVG and MPG mice was not significant (P0.05). In the experiment of space exploration, the number of crossing platforms in MG mice was significantly decreased compared with CG (P0.05). Compared with MG, the number of crossing platforms of MVG and MPG mice increased significantly (P0.05). There was no significant difference in the number of crossing platforms of MVG and MPG mice (P0.05). There was no significant difference in the number of crossing platforms in CG and VG mice (P0.05). The expression of cytochrome C in the CA1 region and the D G region of MG was significantly increased, and the difference was significant (P0.05). Compared with MG, the expression of cytochrome C in the MVG and MPG mice was significantly reduced, and there was a statistical difference (P0.05). There was no significant difference in the expression of cytochrome C in the CA1 and DG regions of the hippocampus between CG and VG (P0.05), and the difference between MVG and MPG was not significant (P0.05). There was a significant difference in the expression of caspase-3 in MVG and MPG mice (P0.05). Compared with CG, the number of positive cells in the dentate gyrus of MG was significantly decreased, and the difference was significant (P0.05); the expression of nestin in MG and MPG was significantly higher than that of MG (P0.05); and the difference between CG and VG was not statistically significant (P0.05). Conclusion:1. Prebiotic E and propafenicol can improve the learning and memory ability of manganese-poisoned mice. Manganese poisoning can promote the increase of cytochrome C in the mouse brain, increase the expression of caspase-3, and reduce the learning and memory capacity of the mouse; and 3, the prebiotic E and the probucol can reduce the expression of the cytochrome C and the caspase-3 in the manganese-poisoned mice, and reduce the apoptosis of the nerve cells; and 4, Manganese poisoning can reduce the number of neural stem cells in the brain of the brain, which can increase the number of neural stem cells in the brain of the mice, and the probucol has no effect on the neural stem cells in the brain of the mice. So as to improve the learning and memory capacity of the manganese-poisoned mice, and meanwhile, the prebiotics E has a certain protective effect on the repair of the brain damage of the manganese poisoning by increasing the number of the neural stem cells in the DG region in the brain of the manganese poisoning.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R595
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