低氧訓(xùn)練對大鼠空間學(xué)習(xí)記憶和海馬齒狀回新生細胞數(shù)量的影響
本文選題:低氧訓(xùn)練 + 大鼠; 參考:《上海體育學(xué)院》2011年碩士論文
【摘要】:研究目的 大腦對低氧具有高度敏感性,一定程度的低氧刺激會對機體各系統(tǒng)、器官、組織和細胞的生理、生化機能甚至結(jié)構(gòu)產(chǎn)生一定影響。已有實驗證明,低氧預(yù)適應(yīng)對大腦具有代償性保護作用。本實驗擬通過動物模型,以雄性SD大鼠為實驗對象,利用低氧和運動作為干預(yù)因素,設(shè)定不同的低氧濃度,探討低氧訓(xùn)練對大鼠空間學(xué)習(xí)記憶的影響以及海馬齒狀回新生細胞數(shù)目的影響,為教練員制定合理、高效、安全的低氧訓(xùn)練方案和促進腦健康提供實驗依據(jù)。 研究方法 1.實驗動物及訓(xùn)練方案:實驗動物為健康7周齡雄性SD(Sprague-Dawley)大鼠60只,體重(266.72±14.38)g。適應(yīng)性喂養(yǎng)3天后,將其隨機分為安靜對照組(C)、常氧運動組(E)、15%氧濃度對照組(15C)、15%氧濃度運動組(15E)、11%氧濃度對照組(11C)和11%氧濃度運動組(11E)6個組,每組10只。運動組動物每天在跑臺上運動,速度為12m/min,運動時間為30min。15E組和11E組每天分別放入15%和11%氧濃度的低氧艙中2h,前30min進行跑臺運動,后1.5h靜置在低氧艙中;15C組和11C組每天分別放入氧濃度為15%和11%的低氧艙中2h。低氧訓(xùn)練7天后,進行Morris水迷宮試驗。細胞形態(tài)學(xué)研究實驗動物共計120只,隨機分為7天組和21天組兩個大組,每組60只。兩個大組的分組情況和訓(xùn)練方案同上,在經(jīng)過7天低氧訓(xùn)練后將7天組的動物進行取材,間隔15天后,再將21天組的動物進行相同的取材。注射BrdU (50mg/kg)開始的時間是正式低氧訓(xùn)練第3天,跑臺運動前1小時給予腹腔注射,持續(xù)注射5天,相應(yīng)各組采取同樣處理。各組動物按設(shè)計要求結(jié)束訓(xùn)練后進行麻醉,依次灌注生理鹽水和多聚甲醛進行沖洗固定,取出腦組織,制作冰凍切片,進行免疫熒光組織化學(xué)染色,熒光顯微鏡下計數(shù)海馬齒狀回免疫陽性細胞數(shù)量。 2.行為學(xué)研究:主要是采用經(jīng)典的Morris水迷宮檢測大鼠的空間學(xué)習(xí)和記憶的能力。 3.細胞形態(tài)學(xué)研究:主要采用免疫熒光組織化學(xué)的實驗方法,對大鼠海馬齒狀回新生細胞數(shù)目進行定量分析。 研究結(jié)果 1.低氧暴露對大鼠空間學(xué)習(xí)記憶能力的影響 1.1低氧暴露對大鼠潛伏期的影響 定位航行試驗中,大鼠尋找隱藏在水面下平臺的時間(潛伏期,latency)反應(yīng)了大鼠空間學(xué)習(xí)的能力。三組大鼠尋找平臺的潛伏期隨訓(xùn)練次數(shù)的增多呈逐漸下降的趨勢,且第二、三天訓(xùn)練的潛伏期與第一天潛伏期比較存在顯著性差異(P0.05)。在第二天訓(xùn)練時,與C組相比,11C組和15C組大鼠的潛伏期明顯低于C組(P0.05),其中15C組大鼠的潛伏期最短。 1.2低氧暴露對大鼠跨平臺次數(shù)的影響 在進行空間探索試驗時,將原有平臺撤除,各組大鼠穿越原有平臺所在位置的次數(shù)反應(yīng)了大鼠的空間記憶能力。在空間探索試驗中,與C組相比,11C組和15C組大鼠跨平臺的次數(shù)均大于C組,其中15C組大鼠跨平臺次數(shù)顯著大于C組(P0.05)。 2.低氧運動對大鼠空間學(xué)習(xí)記憶能力的影響 2.1低氧運動對大鼠潛伏期的影響 在第二天訓(xùn)練中,11E組和15E組大鼠的潛伏期均高于E組,其中11E組大鼠的潛伏期顯著高于E組(Y0.05)。 2.2低氧運動對大鼠跨平臺次數(shù)的影響 空間探索試驗中,11E組和15E組大鼠跨平臺次數(shù)均大于E組,其中15E組大鼠跨平臺的次數(shù)顯著高于E組(P0.05)。 3.低氧暴露對大鼠海馬齒狀回新生細胞數(shù)量的影響 免疫熒光染色結(jié)果顯示,7天11C組和7天15C組的大鼠海馬齒狀回BrdU免疫陽性細胞數(shù)均大于7天C組,但不具有統(tǒng)計學(xué)意義(P0.05),其中7天11C組的大鼠海馬齒狀回BrdU免疫陽性細胞數(shù)最多。間隔15天后,新生細胞存活率分別為:C組61.47%,11C組58.5%,15C組67.65%。 4.低氧運動對大鼠海馬齒狀回新生細胞數(shù)量的影響 免疫熒光染色結(jié)果顯示,7天11E組和7天15E組的大鼠海馬齒狀回BrdU免疫陽性細胞數(shù)均少于7天E組,但不具有統(tǒng)計學(xué)意義(P0.05),其中7天11E組的大鼠海馬齒狀回BrdU免疫陽性細胞數(shù)最少。間隔15天后,新生細胞存活率分別為:E組64.15%,11E組70%,15E組62.05%。 研究結(jié)論 1.低氧刺激可以提高大鼠空間學(xué)習(xí)和記憶能力,其中15%氧濃度的刺激能更加明顯的提高大鼠的空間學(xué)習(xí)和記憶能力。 2.低氧和運動的雙重刺激,對于大鼠的空間學(xué)習(xí)能力具有抑制的作用,同時氧濃度越低,抑制作用越大;然而對于大鼠的空間記憶能力具有促進作用,其中15%氧濃度和運動的雙重刺激更加明顯的提高大鼠的空間記憶能力。 3.低氧刺激可以促進大鼠海馬齒狀回新生細胞的發(fā)生,其中11%氧濃度低氧刺激,效果更加明顯,但是較低的氧濃度不利于新生細胞存活。 4.低氧和運動的雙重刺激抑制大鼠海馬齒狀回新生細胞的發(fā)生,同時氧濃度越低,抑制作用越大,但是較低的氧濃度有利于新生細胞存活。
[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 鹵 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 .
【學(xué)位授予單位】:上海體育學(xué)院
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2011
【分類號】:R363
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