纳洛酮对应激状态下亚急性酒精中毒大鼠学习记忆功能的影响

发布时间：2018-04-03 09:42

本文选题：应激　切入点：亚急性酒精中毒模型　出处：《皖南医学院》2014年硕士论文

【摘要】：目的：观察纳洛酮对应激状态下亚急性酒精中毒大鼠模型影响大鼠学习记忆功能的干预作用。方法：62只雄性SD大鼠随机分为6组：空白对照组（C, n=11），生理盐水组（NS，n=6），纳洛酮组（N，n=11），应激组（St，n=11），亚急性酒精中毒组（SA，n=11），应激后亚急性酒精中毒组（St+SA，n=12）。适应性饲养后开始建立模型，将St组和St+SA组大鼠放入电击箱适应1天后开始给予“声音-电击”配对刺激，连续3天，第5天将大鼠放入电击箱不给予任何刺激，只检测大鼠的木僵反应时间；应激后给予SA组和St+SA组大鼠进行酒精灌胃10天，剂量为15ml/kg，灌胃酒精第1天给予5%浓度酒精溶液，之后在此基础上每天增加5%浓度，第7天后酒精浓度保持为35%；以同样的剂量给予NS组大鼠进行生理盐水灌胃。使用恐惧监测系统检测大鼠的应激状态，使用旷场试验检测大鼠摄入酒精前后的活动度和运动功能的协调度，并使用Morris水迷宫检测纳洛酮对应激状态下亚急性酒精中毒模型大鼠影响学习记忆功能的干预作用。结果： 1.恐惧监测结果分析应激前后、酒精灌胃前后和纳洛酮干预后各组大鼠恐惧检测数据显示，测试日间F（3，168）=12.53，P0.001，处理组间F（5，56）=13.60，P0.001； SNK-q两两比较：St组和St+SA组与其他四组之间的恐惧监测都存在着显著性差异（P0.05）。首先，应激前后6组大鼠恐惧监测数据的比较：St组（t=-8.28，P0.001）与St+SA组（t=-6.62，P0.001）都具有显著性差异。应激后C组与St组（t=-7.52，P0.001）、C组与St+SA组（t=-5.19，，P0.001）、SA组与St+SA组之间也具有显著性差异（t=-2.43，P0.05）。其次，酒精灌胃前后6组大鼠恐惧监测数据的比较：N组（t=-3.89，P0.001）、St组（t=4.48，P0.001）、St+SA组（t=2.27，P0.05）具有显著性差异；C组与NS组大鼠在酒精灌胃前（t=-0.12，P0.05）和酒精灌胃后（t=-0.56，P0.05）均无显著性差异。最后，纳洛酮干预后6组大鼠恐惧监测数据相比F=3.50，P0.01；N组、St组和St+SA组分别与其他组相比P0.05。 2.旷场试验结果分析 6组大鼠酒精灌胃后的旷场试验中央格停留时间（F=2.36，P0.05）、总穿格数（F=7.04，P0.001）、站立次数（F=8.56，P0.001）都具有显著性差异，修饰次数和粪便粒数（P0.05）无显著性差异；采用SNK-q两两比较，总穿格数C组与其他5组之间P0.05， St组与其他4组（不包括N组）之间P0.05，SA组与其他5组之间P0.05，St+SA组与其他5组之间P0.05；站立次数C组和St组分别与SA组和St+SA组之间P0.05。6组大鼠酒精灌胃前后的旷场试验中总穿格数（t=9.57，P0.001）、站立次数（t=6.85，P0.001）、修饰次数（t=-4.93，P0.001）、粪便粒数（t=2.10，P0.05）都具有显著性差异。C组与SA组大鼠在酒精灌胃前后旷场试验的中央格停留时间（t=3.31，P0.01）、总穿格数（t=2.60，P0.05）和站立次数（t=3.49，P0.01）均具有显著性差异。 3. Morris水迷宫测试结果分析采用双因素方差分析（重复测量）对大鼠行为Morris水迷宫训练定位航行阶段7天的逃避潜伏期进行分析，测试日间F（6，336）=23.15，P0.001；处理组间F（5，56）=1.34，P0.05；测试日间与组别的交互作用F（1，56）=594.84，P0.001；且NS组与St+SA组之间差异有统计学意义（P0.05）。采用单因素方差分析对每组大鼠7天的行为Morris水迷宫训练定位航行阶段的逃避潜伏期进行分析，C组（F=6.19，P0.001），NS组（F=4.77，P0.001），N组（F=6.96，P0.001），St组（F=4.77，P0.001），SA组（F=1.14，P0.05），St+SA组（F=3.32，P0.01）。定位航行阶段第4天C组与SA组大鼠的逃避潜伏期之间具有显著性差异（t=2.32，P0.05）。通过卡方检验对大鼠7天定位航行阶段的分析策略构成比进行分析，Day1(χ2=867.20，P0.001)，Day2(χ2=898.70，P0.001)，Day3(χ2=666.30，P0.001)，Day4(χ2=825.70，P0.001)，Day5(χ2=108.90，P0.001)，Day6(χ2=932.80，P0.001)，Day7(χ2=675.40，P0.001)。采用双因素方差分析（重复测量）对大鼠行为Morris水迷宫训练定位航行阶段7天的游泳速度进行分析，测试日间（F=19.19，P0.001），在定位航行阶段第5天处理组间（F=3.25，P0.01），且在整个定位航行阶段中各组的游泳速度呈下降趋势；第6天C组与SA组之间（F=-2.68，P0.05），第5天C组与St+SA组具有显著性差异（F=2.45，P0.05）。比较大鼠在定位航行阶段的靶象限活动时间百分比，第4天C组分别与SA组（t=2.13，P0.05）、St+SA组（t=2.18，P0.05）之间差异具有统计学意义；第6天NS组与N组之间差异显著（t=-2.25，P0.05）。 NS组与N组大鼠在空间探索阶段的穿台次数差异具有统计学意义（t=-2.34，P0.05）。 4.各组大鼠体重比较采用双因素方差分析（重复测量）对各组大鼠在应激前后、酒精灌胃前后和纳洛酮干预后的体重进行分析，与C组相比，模型组大鼠的体重增长明显受抑制（P0.05），并且各处理组间相比F（5，56）=11.71，P0.001；测试日间F（4，224）=213.80，P0.001。使用SNK-q进行两两比较， SA组和St+SA组与其他组相比P0.05，而SA组和St+SA组等之间的体重无显著性差异（P0.05）。结论酒精对大鼠的活动度和体重增长等具有抑制作用；应激状态下亚急性酒精中毒可损害大鼠学习记忆功能；纳洛酮对应激状态下亚急性酒精中毒大鼠学习记忆功能有一定的改善作用。
[Abstract]:Objective : To observe the effect of naloxone on learning and memory function in rats with subacute alcoholism .

Methods : 62 male SD rats were randomly divided into six groups : blank control group ( C , n = 11 ) , physiological saline group ( NS , n = 6 ) , naloxone group ( N , n = 11 ) , stress group ( St , n = 11 ) , subacute alcoholism group ( SA , n = 11 ) , post - stress subacute alcoholism group ( St + SA , n = 12 ) . After adaptive feeding , the model was established , and the rats in the St and St + SA groups were put into the electric shock box for 1 day , then the " voice - electric shock " pairing was started , and the rats were put into the electric shock box for 3 days , and the rats were put into the electric shock box without any stimulation , and the reaction time of the rats was detected only ;
After stress , the rats were given SA group and St + SA group for 10 days , the dosage was 15ml / kg , the alcohol solution of 5 % concentration was given on the first day of alcohol administration , then the concentration of alcohol was increased by 5 % and the alcohol concentration was kept at 35 % after 7 days .
In the same dose , NS group rats were given normal saline enema . Using the fear monitoring system to detect the stress state of rats , the activity degree and the coordination degree of motion function before and after alcohol intake were detected by open field test , and the effect of naloxone on learning and memory function was studied by Morris water maze test in rats with subacute alcoholism model .

Results :

1 . Analysis of the results of fear monitoring

The results showed that F ( 3,168 ) = 12.53 , P0.001 , F ( 5,56 ) = 13.60 , P0.001 ;
Compared with the other four groups ( t = - 8.28 , P0.001 ) and St + SA group ( t = - 6.62 , P0.001 ) , there was significant difference between group C and St group ( t = - 7.52 , P0.001 ) , group C and St + SA group ( t = - 5.19 , P0.001 ) , and there was significant difference between group SA and St + SA group ( t = - 2.43 , P0.05 ) . Secondly , there was significant difference between group N ( t = - 3.89 , P0.001 ) , St group ( t = 4.48 , P0.001 ) , St + SA group ( t = 2.27 , P0.05 ) .
There was no significant difference between group C and NS group ( t = - 0.12 , P0.05 ) and alcohol ( t = - 0.56 , P0.05 ) .
Compared with other groups , the group N , St and St + SA were significantly higher than those in other groups ( P0.05 ) .

2 . Analysis of open field test results

The time ( F = 2.36 , P < 0.01 ) and the number of standing ( F = 8.56 , P0.001 ) were significantly different between the six groups ( F = 2.36 , P < 0.01 ) .
Compared with other five groups , there was no significant difference between the group C and the other 5 groups ( P0.05 ) .
There was significant difference between group C and group SA ( t = 9.57 , P0.001 ) , number of standing ( t = 6.85 , P0.001 ) , number of days ( t = - 4.93 , P0.001 ) , number of feces ( t = - 4.93 , P0.001 ) , number of stool grains ( t = - 4.93 , P0.05 ) , and the number of standing ( t = 2.60 , P0.05 ) and standing frequency ( t = 3.49 , P0.01 ) .

3 . Analysis of Morris water maze test results

Two - factor variance analysis ( repeated measurement ) was used to analyze the escape latency of Morris water maze training for 7 days in Morris water maze training , and the daytime F ( 6,336 ) = 23.15 , P0.001 ;
F ( 5 , 56 ) = 1 . 34 , P 0 . 05 in treatment group ;
F ( 1 , 56 ) = 594.84 , P0.001 ;
In group C ( F = 6.19 , P0.001 ) , NS group ( F = 4.77 , P0.001 ) , NS group ( F = 4.77 , P0.001 ) , group N ( F = 6.96 , P0.001 ) , St group ( F = 4.77 , P0.001 ) , SA group ( F = 1.14 , P0.05 ) , St + SA group ( F = 3.32 , P0.01 ) . There was a significant difference ( t = 2.32 , P < 0.05 ) between group C and SA group ( t = 2.32 , P0.05 ) .

Day1 ( 蠂2 = 867.20 , P0.001 ) , Day2 ( 蠂2 = 898.70 , P0.001 ) , Day3 ( 蠂2 = 867.30 , P0.001 ) , Day5 ( 蠂2 = 108.90 , P0.001 ) , Day6 ( 蠂2 = 932.80 , P0.001 ) , Day7 ( 蠂2 = 670.40 , P0.001 ) .

Two - factor analysis ( repeated measurement ) was used to analyze the swimming speed of rats ' behavior Morris water maze training for 7 days , and the daytime ( F = 19.19 , P0.001 ) was measured , and the swimming speed of each group was decreased during the fifth day of the navigation phase ( F = 3.25 , P0.01 ) .
There was significant difference between group C and SA group ( F = - 2.68 , P0.05 ) .

Compared with SA group ( t = 2.13 , P 0.05 ) and St + SA group ( t = 2.18 , P 0.05 ) , the percentage of active time of target quadrant was compared with that of group SA ( t = 2.13 , P0.05 ) .
There was a significant difference between NS group and N group on Day 6 ( t = - 2.25 , P0.05 ) .

There was significant difference between NS group and N group ( t = - 2.34 , P0.05 ) .

4 . Body weight comparison in each group

Compared with group C , the weight gain of model group rats was significantly inhibited compared with group C ( P0.05 ) , and F ( 5,56 ) = 11.71 , P0.001 ;
Compared with other groups , there was no significant difference between SA group and St + SA group ( P0.05 ) .

Conclusion Alcohol has an inhibitory effect on the activity and body weight gain of rats .
Subacute alcoholism could damage the learning and memory function of rats in stress state .
The effects of naloxone on learning and memory function in rats with subacute alcoholism were improved .

【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：B842.3

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