本文選題：恐傷母鼠 + 神經突觸�。� 參考：《河南中醫(yī)學院》2012年碩士論文

【摘要】：目的 采用旁觀電擊法、危境求生法以及模擬地震法制備慢性單純的恐傷母鼠動物模型,通過觀察母鼠的一般情況、孕產情況以及仔鼠生長發(fā)育情況,檢測仔鼠空間學習和記憶能力以及海馬區(qū)形態(tài)學、分子生物學變化,篩選出最佳的造模方法,并初步探討恐傷母鼠對其仔鼠空間學習和記憶能力的影響及其發(fā)生的分子機制。 方法 2月齡SPF級Wistar大鼠,體重180±20g,其中雌鼠60只,雄鼠40只。按隨機數字表法把60只雌鼠分為旁觀電擊組、危境求生組、模擬地震組和空白對照組,每組15只。從40只雄鼠中選出體重較大的30只作為交配鼠,剩余10只作為電擊鼠。旁觀電擊組自造模第1d起于9AM將電擊鼠和旁觀鼠放入自制交流箱相應的小室內,電擊10次/d,3min/次；危境求生組自造模第1d起于9AM將求生鼠放入的自制危境箱內,1次/d,30min/次；模擬地震組自造模第1d起于9AM將其置于氣浴振蕩器內,10次/d,3min/次。第15d起于9PM按雌雄2：1合籠,次日7AM檢查受孕情況,造模持續(xù)到孕第19d。造模期間空白對照組雌鼠置于安靜的環(huán)境內30min。母鼠生產后每組抽取10窩每窩8只仔鼠進行喂養(yǎng)和實驗。每組每窩仔鼠于第14、21d分別隨機取出2只,解剖取海馬組織以備檢測,每組每窩仔鼠于1月齡分別取出2只進行Morris水迷宮實驗。觀察及檢測指標：(1)母鼠的一般情況和孕產情況；(2)仔鼠的翻身、出牙等生長發(fā)育情況；(3)在電鏡下觀察14日齡、21日齡的仔鼠海馬區(qū)神經細胞突觸結構的改變；(4)采用免疫組化法檢測21日齡仔鼠海馬CA3區(qū)的p-CREB、SYN-1的蛋白表達情況。(5)應用Morris水迷宮實驗測試1月齡仔鼠空間學習和記憶能力。 結果 1恐傷母鼠動物模型的制備：成功制備三種慢性單純的恐傷母鼠動物模型。 2恐傷母鼠一般狀況的變化：與空白對照組比較,模型組雌鼠皮毛色澤情況、反應敏捷度、活動靈活度、精神狀態(tài)都明顯的異常,按旁觀電擊組、危境求生組、模擬地震組順序異常程度依次減弱。 3母鼠的孕產情況：與空白對照組比較,模型組的產仔數減少,差別均有統(tǒng)計學意義(P0.05)；但各模型組產仔數,孕鼠數以及產仔孕鼠數各組間的差別均無統(tǒng)計學意義(P0.05)。 4恐傷母鼠其仔鼠的生長情況：與空白對照組比較,模型組的仔鼠翻身、出牙和開眼時間都延長,差別均有統(tǒng)計學意義(P0.05)；模型組間比較,差別均無統(tǒng)計學意義(P0.05)。 5恐傷母鼠其仔鼠海馬區(qū)神經細胞突觸結構的改變：模型組仔鼠海馬區(qū)腦神經細胞突觸小泡、線粒體都有不同程度的改變,按旁觀電擊組、危境求生組、模擬地震組順序改變程度依次減弱。 6恐傷母鼠其21日齡仔鼠海馬CA3區(qū)p-CREB.SYN-1蛋白表達情況：與空白對照組比較,旁觀電擊組、危境求生組兩種蛋白表達減弱,差別均有統(tǒng)計學意義(P0.05),模擬地震組的差別均無統(tǒng)計學意義(P0.05)；與旁觀電擊組比較,模擬地震組兩種蛋白表達增強差別均有統(tǒng)計學意義(P0.05),危境求生組的差別均無統(tǒng)計學意義(P0.05)；與危境求生組比較,模擬地震組兩種蛋白表達增強,差別均有統(tǒng)計學意義(P0.05)。 7恐傷母鼠對其1月齡仔鼠空間學習和記憶能力的影響：(1)與空白對照組比較,旁觀電擊組4天的平均潛伏期均延長,差別有統(tǒng)計學意義(P0.05),危境求生組和模擬地震組第1天的平均潛伏期延長,其余三天縮短,差別無統(tǒng)計學意義(P0.05)；與旁觀電擊組比較,各組4天的平均潛伏期均縮短,差別有統(tǒng)計學意義(P0.05)；危境求生組與模擬地震組比較,4天的平均潛伏期差別無統(tǒng)計學意義(P0.05)。(2)組間的搜索策略差異有統(tǒng)計學意義(x2=5.208,P=0.042)；與空白對照組比較,旁觀電擊組仔鼠較多采用非有效策略,較少采用有效策略方式的搜索策略,差異有統(tǒng)計學意義(x2=5.104,P=0.024),其余各組間的搜索策略差異無統(tǒng)計學意義(P0.05)；(3)與空白對照組比較,旁觀電擊組仔鼠平均游泳速度減慢、在20%邊緣區(qū)平均百分比增大,在其他區(qū)域平均百分比減小,差異均有統(tǒng)計學意義(P0.05)；模擬地震組平均游泳速度減慢,差異有統(tǒng)計學意義(P0.05),其余差異均無統(tǒng)計學意義(P0.05)；與旁觀電擊組比較,危境求生組平均游泳速度、在20%邊緣區(qū)還有其他區(qū)域的差異均有統(tǒng)計學意義(P0.05),模擬地震組差異均無統(tǒng)計學意義(P0.05)；危境求生組與模擬地震組的差異均無統(tǒng)計學意義(P0.05) 結論 1三種慢性單純的恐傷腎動物造模方法中,旁觀電擊法制備的模型最適合。 2恐傷母鼠不僅可影響自身的產仔情況,還可導致仔鼠空間學習和記憶能力降低以及海馬區(qū)神經細胞突觸和蛋白表達的改變。 3恐傷母鼠對其仔鼠空間學習和記憶能力以及海馬神經突觸的影響其可能機制：恐傷母鼠可使仔鼠海馬CA3區(qū)的p-CERB數量減少,從而引起下游SYN-1的表達減少,進而影響海馬神經突觸的改變,最終導致海馬區(qū)形態(tài)結構和功能的異常。
[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 鹵 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 .
【學位授予單位】：河南中醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R749.7

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