恐伤母鼠对其仔鼠空间学习记忆能力的影响及其神经发生机制

发布时间：2018-07-08 08:20

本文选题：恐伤母鼠 + 神经突触　；参考：《河南中医学院》2012年硕士论文

【摘要】：目的采用旁观电击法、危境求生法以及模拟地震法制备慢性单纯的恐伤母鼠动物模型,通过观察母鼠的一般情况、孕产情况以及仔鼠生长发育情况,检测仔鼠空间学习和记忆能力以及海马区形态学、分子生物学变化,筛选出最佳的造模方法,并初步探讨恐伤母鼠对其仔鼠空间学习和记忆能力的影响及其发生的分子机制。方法 2月龄SPF级Wistar大鼠,体重180±20g,其中雌鼠60只,雄鼠40只。按随机数字表法把60只雌鼠分为旁观电击组、危境求生组、模拟地震组和空白对照组,每组15只。从40只雄鼠中选出体重较大的30只作为交配鼠,剩余10只作为电击鼠。旁观电击组自造模第1d起于9AM将电击鼠和旁观鼠放入自制交流箱相应的小室内,电击10次/d,3min/次；危境求生组自造模第1d起于9AM将求生鼠放入的自制危境箱内,1次/d,30min/次；模拟地震组自造模第1d起于9AM将其置于气浴振荡器内,10次/d,3min/次。第15d起于9PM按雌雄2：1合笼,次日7AM检查受孕情况,造模持续到孕第19d。造模期间空白对照组雌鼠置于安静的环境内30min。母鼠生产后每组抽取10窝每窝8只仔鼠进行喂养和实验。每组每窝仔鼠于第14、21d分别随机取出2只,解剖取海马组织以备检测,每组每窝仔鼠于1月龄分别取出2只进行Morris水迷宫实验。观察及检测指标：(1)母鼠的一般情况和孕产情况；(2)仔鼠的翻身、出牙等生长发育情况；(3)在电镜下观察14日龄、21日龄的仔鼠海马区神经细胞突触结构的改变；(4)采用免疫组化法检测21日龄仔鼠海马CA3区的p-CREB、SYN-1的蛋白表达情况。(5)应用Morris水迷宫实验测试1月龄仔鼠空间学习和记忆能力。结果 1恐伤母鼠动物模型的制备：成功制备三种慢性单纯的恐伤母鼠动物模型。 2恐伤母鼠一般状况的变化：与空白对照组比较,模型组雌鼠皮毛色泽情况、反应敏捷度、活动灵活度、精神状态都明显的异常,按旁观电击组、危境求生组、模拟地震组顺序异常程度依次减弱。 3母鼠的孕产情况：与空白对照组比较,模型组的产仔数减少,差别均有统计学意义(P0.05)；但各模型组产仔数,孕鼠数以及产仔孕鼠数各组间的差别均无统计学意义(P0.05)。 4恐伤母鼠其仔鼠的生长情况：与空白对照组比较,模型组的仔鼠翻身、出牙和开眼时间都延长,差别均有统计学意义(P0.05)；模型组间比较,差别均无统计学意义(P0.05)。 5恐伤母鼠其仔鼠海马区神经细胞突触结构的改变：模型组仔鼠海马区脑神经细胞突触小泡、线粒体都有不同程度的改变,按旁观电击组、危境求生组、模拟地震组顺序改变程度依次减弱。 6恐伤母鼠其21日龄仔鼠海马CA3区p-CREB.SYN-1蛋白表达情况：与空白对照组比较,旁观电击组、危境求生组两种蛋白表达减弱,差别均有统计学意义(P0.05),模拟地震组的差别均无统计学意义(P0.05)；与旁观电击组比较,模拟地震组两种蛋白表达增强差别均有统计学意义(P0.05),危境求生组的差别均无统计学意义(P0.05)；与危境求生组比较,模拟地震组两种蛋白表达增强,差别均有统计学意义(P0.05)。 7恐伤母鼠对其1月龄仔鼠空间学习和记忆能力的影响：(1)与空白对照组比较,旁观电击组4天的平均潜伏期均延长,差别有统计学意义(P0.05),危境求生组和模拟地震组第1天的平均潜伏期延长,其余三天缩短,差别无统计学意义(P0.05)；与旁观电击组比较,各组4天的平均潜伏期均缩短,差别有统计学意义(P0.05)；危境求生组与模拟地震组比较,4天的平均潜伏期差别无统计学意义(P0.05)。(2)组间的搜索策略差异有统计学意义(x2=5.208,P=0.042)；与空白对照组比较,旁观电击组仔鼠较多采用非有效策略,较少采用有效策略方式的搜索策略,差异有统计学意义(x2=5.104,P=0.024),其余各组间的搜索策略差异无统计学意义(P0.05)；(3)与空白对照组比较,旁观电击组仔鼠平均游泳速度减慢、在20%边缘区平均百分比增大,在其他区域平均百分比减小,差异均有统计学意义(P0.05)；模拟地震组平均游泳速度减慢,差异有统计学意义(P0.05),其余差异均无统计学意义(P0.05)；与旁观电击组比较,危境求生组平均游泳速度、在20%边缘区还有其他区域的差异均有统计学意义(P0.05),模拟地震组差异均无统计学意义(P0.05)；危境求生组与模拟地震组的差异均无统计学意义(P0.05) 结论 1三种慢性单纯的恐伤肾动物造模方法中,旁观电击法制备的模型最适合。 2恐伤母鼠不仅可影响自身的产仔情况,还可导致仔鼠空间学习和记忆能力降低以及海马区神经细胞突触和蛋白表达的改变。 3恐伤母鼠对其仔鼠空间学习和记忆能力以及海马神经突触的影响其可能机制：恐伤母鼠可使仔鼠海马CA3区的p-CERB数量减少,从而引起下游SYN-1的表达减少,进而影响海马神经突触的改变,最终导致海马区形态结构和功能的异常。
[Abstract]:Purpose

By observing the general situation of mother and mouse , the situation of pregnancy and the development of newborn rats , the study on the study of space learning and memory and the morphological and molecular biology of the hippocampus were carried out by observing the general situation of the mother mouse , the changes of molecular biology and the selection of the best modeling method .

method

Two - month - old SPF Wistar rats weighing 180 卤 20g , including 60 female rats and 40 male rats , were randomly divided into two groups : one electric shock group , a critical survival group , an analog seismic group and a blank control group .
From the first day of the model day 1d , the survival group was put into a self - made critical box , 1 time / d , 30 min / time .
The rats were divided into 2 : 1 cages at 9 PM and 7 AM on the next day for 30 min .
( 2 ) the growth and development of newborn mice , such as turning over and out of teeth , etc . ;
( 3 ) Under electron microscope , the synaptic structure of neural cells was observed in the hippocampus of 14 - day - old and 21 - day - old mice .
( 4 ) The expression of p - CREB and SYN - 1 was detected by immunohistochemistry . ( 5 ) The spatial learning and memory abilities were measured by Morris water maze test .

Results

1 . Preparation of animal models for fear of injury : Three types of chronic simple fear - wound mother - mouse models were successfully prepared .

2 In comparison with blank control group , the abnormal degree of fur color , agility , activity flexibility and mental state of the model group were obviously abnormal , and the degree of abnormal earthquake group was decreased in sequence .

Compared with the blank control group , the litter size of the model group was decreased and the difference was statistically significant ( P0.05 ) .
However , there was no significant difference between the number of litter , the number of pregnant rats and the number of pregnant rats in each model group ( P0.05 ) .

The results showed that the growth of newborn mice was significantly higher than that in the control group ( P0.05 ) .
There was no significant difference between the model groups ( P0.05 ) .

5 . The synaptic structure of neural cells in the hippocampus area of neonatal rats was changed : the synaptic vesicle and mitochondria of the brain nerve cells in the hippocampus of the model group were changed to some extent , and the degree of change of the sequence of the simulated seismic group was decreased in sequence according to the observation group and the critical survival group .

The expression of p - CREB . SYN - 1 protein in hippocampus CA 3 region of neonatal rats with 6 months old was significantly different from that in control group ( P0.05 ) , but there was no significant difference between the two groups ( P0.05 ) .
Compared with the observation group , the difference of expression of two proteins in the simulated earthquake group was significant ( P0.05 ) , and the difference between the two groups was not statistically significant ( P0.05 ) .
Compared with the critical survival group , the expression of two proteins in the simulated earthquake group was enhanced , and the difference was statistically significant ( P0.05 ) .

( 1 ) Compared with the blank control group , the average latency of 4 days was prolonged , the difference was significant ( P0.05 ) , the average latency in the critical survival group and the simulated earthquake group was prolonged , the remaining three days were shortened , the difference was not significant ( P0.05 ) ;
Compared with the observation group , the average latency of 4 days in each group was shorter and the difference was significant ( P0.05 ) .
There was no significant difference in the average latency between groups ( P 0.05 ) . The difference of search strategy between groups was significant ( x2 = 5.208 , P = 0.042 ) .
Compared with the blank control group , compared with the blank control group , there was a significant difference in the search strategy ( x2 = 5.104 , P = 0 . 024 ) , and there was no significant difference in the search strategy among the rest groups ( P0.05 ) .
( 3 ) Compared with the blank control group , the average swimming speed was slow , the average percentage of the 20 % margin area increased , the average percentage of other regions decreased , the difference was statistically significant ( P0.05 ) ;
The average swimming speed of the simulated earthquake group was slow , the difference was significant ( P0.05 ) , and the difference was not statistically significant ( P0.05 ) .
Compared with the observation group , the average swimming speed of the critical survival group and the difference of the other regions in the 20 % margin area were statistically significant ( P0.05 ) , and the difference of the simulated seismic group was not statistically significant ( P0.05 ) ;
There was no significant difference between the critical survival group and the simulated earthquake group ( P0.05 ) .

Conclusion

One of the three models of chronic simple fear - induced injury of the kidney is the most suitable model for the preparation of the side - view electric shock method .

In addition , it can not only affect the litter size , but also result in the decrease of space learning and memory ability and the change of synaptic and protein expression in neural cells in hippocampus .

3 . The possible mechanism of damage to the space learning and memory ability of newborn mice and the synaptic transmission in hippocampus is that the number of p - CERB in hippocampus CA 3 region of newborn rats can be decreased , which can reduce the expression of SYN - 1 in hippocampus , which can affect the changes of synaptic changes in hippocampus , and eventually lead to abnormal morphological structure and function of hippocampus .
【学位授予单位】：河南中医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R749.7

【参考文献】