低氧训练对大鼠空间学习记忆和海马齿状回新生细胞数量的影响

发布时间：2018-06-04 08:32

本文选题：低氧训练 + 大鼠　；参考：《上海体育学院》2011年硕士论文

【摘要】：研究目的大脑对低氧具有高度敏感性,一定程度的低氧刺激会对机体各系统、器官、组织和细胞的生理、生化机能甚至结构产生一定影响。已有实验证明,低氧预适应对大脑具有代偿性保护作用。本实验拟通过动物模型,以雄性SD大鼠为实验对象,利用低氧和运动作为干预因素,设定不同的低氧浓度,探讨低氧训练对大鼠空间学习记忆的影响以及海马齿状回新生细胞数目的影响,为教练员制定合理、高效、安全的低氧训练方案和促进脑健康提供实验依据。研究方法 1.实验动物及训练方案：实验动物为健康7周龄雄性SD(Sprague-Dawley)大鼠60只,体重(266.72±14.38)g。适应性喂养3天后,将其随机分为安静对照组(C)、常氧运动组(E)、15%氧浓度对照组(15C)、15%氧浓度运动组(15E)、11%氧浓度对照组(11C)和11%氧浓度运动组(11E)6个组,每组10只。运动组动物每天在跑台上运动,速度为12m/min,运动时间为30min。15E组和11E组每天分别放入15%和11%氧浓度的低氧舱中2h,前30min进行跑台运动,后1.5h静置在低氧舱中；15C组和11C组每天分别放入氧浓度为15%和11%的低氧舱中2h。低氧训练7天后,进行Morris水迷宫试验。细胞形态学研究实验动物共计120只,随机分为7天组和21天组两个大组,每组60只。两个大组的分组情况和训练方案同上,在经过7天低氧训练后将7天组的动物进行取材,间隔15天后,再将21天组的动物进行相同的取材。注射BrdU (50mg/kg)开始的时间是正式低氧训练第3天,跑台运动前1小时给予腹腔注射,持续注射5天,相应各组采取同样处理。各组动物按设计要求结束训练后进行麻醉,依次灌注生理盐水和多聚甲醛进行冲洗固定,取出脑组织,制作冰冻切片,进行免疫荧光组织化学染色,荧光显微镜下计数海马齿状回免疫阳性细胞数量。 2.行为学研究：主要是采用经典的Morris水迷宫检测大鼠的空间学习和记忆的能力。 3.细胞形态学研究：主要采用免疫荧光组织化学的实验方法,对大鼠海马齿状回新生细胞数目进行定量分析。研究结果 1.低氧暴露对大鼠空间学习记忆能力的影响 1.1低氧暴露对大鼠潜伏期的影响定位航行试验中,大鼠寻找隐藏在水面下平台的时间(潜伏期,latency)反应了大鼠空间学习的能力。三组大鼠寻找平台的潜伏期随训练次数的增多呈逐渐下降的趋势,且第二、三天训练的潜伏期与第一天潜伏期比较存在显著性差异(P0.05)。在第二天训练时,与C组相比,11C组和15C组大鼠的潜伏期明显低于C组(P0.05),其中15C组大鼠的潜伏期最短。 1.2低氧暴露对大鼠跨平台次数的影响在进行空间探索试验时,将原有平台撤除,各组大鼠穿越原有平台所在位置的次数反应了大鼠的空间记忆能力。在空间探索试验中,与C组相比,11C组和15C组大鼠跨平台的次数均大于C组,其中15C组大鼠跨平台次数显著大于C组(P0.05)。 2.低氧运动对大鼠空间学习记忆能力的影响 2.1低氧运动对大鼠潜伏期的影响在第二天训练中,11E组和15E组大鼠的潜伏期均高于E组,其中11E组大鼠的潜伏期显著高于E组(Y0.05)。 2.2低氧运动对大鼠跨平台次数的影响空间探索试验中,11E组和15E组大鼠跨平台次数均大于E组,其中15E组大鼠跨平台的次数显著高于E组(P0.05)。 3.低氧暴露对大鼠海马齿状回新生细胞数量的影响免疫荧光染色结果显示,7天11C组和7天15C组的大鼠海马齿状回BrdU免疫阳性细胞数均大于7天C组,但不具有统计学意义(P0.05),其中7天11C组的大鼠海马齿状回BrdU免疫阳性细胞数最多。间隔15天后,新生细胞存活率分别为：C组61.47%,11C组58.5%,15C组67.65%。 4.低氧运动对大鼠海马齿状回新生细胞数量的影响免疫荧光染色结果显示,7天11E组和7天15E组的大鼠海马齿状回BrdU免疫阳性细胞数均少于7天E组,但不具有统计学意义(P0.05),其中7天11E组的大鼠海马齿状回BrdU免疫阳性细胞数最少。间隔15天后,新生细胞存活率分别为：E组64.15%,11E组70%,15E组62.05%。研究结论 1.低氧刺激可以提高大鼠空间学习和记忆能力,其中15%氧浓度的刺激能更加明显的提高大鼠的空间学习和记忆能力。 2.低氧和运动的双重刺激,对于大鼠的空间学习能力具有抑制的作用,同时氧浓度越低,抑制作用越大；然而对于大鼠的空间记忆能力具有促进作用,其中15%氧浓度和运动的双重刺激更加明显的提高大鼠的空间记忆能力。 3.低氧刺激可以促进大鼠海马齿状回新生细胞的发生,其中11%氧浓度低氧刺激,效果更加明显,但是较低的氧浓度不利于新生细胞存活。 4.低氧和运动的双重刺激抑制大鼠海马齿状回新生细胞的发生,同时氧浓度越低,抑制作用越大,但是较低的氧浓度有利于新生细胞存活。
[Abstract]:Purpose of study

The brain has high sensitivity to hypoxia , and a certain degree of hypoxic stimulation has some effects on the physiological , biochemical functions and even structure of various organs , organs , tissues and cells of the organism . It has been proved that hypoxic preconditioning has compensatory protective effect on the brain . The effect of hypoxia training on the memory of rats ' space learning and the effect of hypoxic training on the number of neonatal cells in hippocampus were studied .

Research Methods

1 . Experimental animals and training regimen : 60 male SD rats ( 262.72 卤 14.38 ) g were randomly divided into two groups : a quiet control group ( C ) , a normal oxygen movement group ( E ) , a 15 % oxygen concentration control group ( 15C ) , a 15 % oxygen concentration exercise group ( E ) , a 15 % oxygen concentration control group ( 15C ) , a 15 % oxygen concentration exercise group ( C ) , a 11 % oxygen concentration control group ( 11C ) , and a 11 % oxygen concentration exercise group .
After 7 days of hypoxic training , 120 rats were randomly divided into 7 day group and 21 day group .

2 . Behavior study : The study of ethology mainly adopts the classical Morris water maze to test the spatial learning and memory capacity of rats .

3 . The morphological study of cells : The quantitative analysis of the number of neonatal rat hippocampal dentate gyrus cells was carried out by using the experimental method of immunofluorescence histochemistry .

Results of the study

1 . Effect of hypoxia exposure on learning and memory ability of rats

1.1 Effects of hypoxia exposure on latency of rats

During the second day of training , the latency of group C and 15C was significantly lower than that in group C ( P0.05 ) , and the latency of group C was the shortest .

1.2 Effect of hypoxia exposure on cross - platform frequency of rats

In the space exploration experiment , the number of cross - platform of group C and group C was more than that in group C ( P0.05 ) .

2 . Effect of hypoxic exercise on learning and memory ability of rats

2.1 Effects of hypoxia on latency of rats

In the second day of training , the latency of the E group was higher than that in E group , and the latency of the model group was significantly higher than that in group E ( Y 0.05 ) .

2.2 Effects of hypoxic exercise on cross - platform frequency of rats

In the space exploration experiment , the number of cross - platform cross - platform was more than that in E group , among which the number of cross - platform was significantly higher than that in group E ( P0.05 ) .

3 . Effect of hypoxia exposure on neonatal rat hippocampal dentate gyrus

Immunofluorescence staining showed that the number of positive cells in dentate gyrus of hippocampus dentate gyrus was more than 7 days in group C and 7 days , but there was no statistical significance ( P0.05 ) . After 15 days , the survival rate of neonatal cells was 61.47 % in group C , 58.5 % in 11C group and 67.65 % in 15C group .

4 . Effect of hypoxic exercise on the number of neonatal rat hippocampal dentate gyrus cells

Immunofluorescence staining showed that the number of positive cells in dentate gyrus of hippocampus dentate gyrus in 7 days and 7 days was less than that in group E , but there was no statistical significance ( P0.05 ) . The survival rates of neonatal cells were 64.15 % , 70 % and 62.05 % in E group after 15 days .

Conclusions of the study

1 . Hypoxia stimulation can improve the spatial learning and memory ability of rats , and 15 % oxygen concentration can improve the spatial learning and memory ability of rats .

2 . The dual stimulation of hypoxia and exercise has an inhibitory effect on the spatial learning ability of rats , and the lower the oxygen concentration , the greater the inhibition effect ;
However , the spatial memory capacity of rats was promoted , and 15 % of the oxygen concentration and the dual stimulation of exercise were more obvious to improve the spatial memory ability of the rats .

3 . Hypoxia stimulation can promote the development of neonatal rat hippocampal dentate gyrus , in which 11 % oxygen concentration is low oxygen stimulation , the effect is more obvious , but lower oxygen concentration is not good for the survival of neonatal cells .

4 . Dual stimulation of hypoxia and exercise inhibited the occurrence of neonatal rat hippocampal dentate gyrus , while the lower the oxygen concentration , the greater the inhibitory effect , but lower oxygen concentration was beneficial to the survival of neonatal cells .
【学位授予单位】：上海体育学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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