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  本文選題：低氧預(yù)適應(yīng)　切入點(diǎn)：缺血再灌注　出處：《泰山醫(yī)學(xué)院》2014年碩士論文

【摘要】：研究目的： 1．研究低氧預(yù)適應(yīng)對缺血再灌注腦損傷小鼠學(xué)習(xí)記憶的影響。 2．研究低氧預(yù)適應(yīng)對缺血再灌注腦損傷后海馬CA1區(qū)神經(jīng)細(xì)胞凋亡的影響。 3．研究低氧預(yù)適應(yīng)對缺血再灌注腦損傷后海馬CA1區(qū)MAP-2表達(dá)的影響。 研究方法： 1．取體重為18-22g健康成年昆明小鼠，雄性，30只，將小鼠隨機(jī)分為正常組（Ngroup）、假手術(shù)組（C group）、缺血再灌注組（O group）、低氧預(yù)適應(yīng)組（HPC group）和低氧預(yù)適應(yīng)+缺血再灌注組（HPC+O group）。每組小鼠6只。應(yīng)用Morris水迷宮實(shí)驗(yàn)進(jìn)行行為學(xué)檢測，記錄定位航行實(shí)驗(yàn)的平均潛伏期及空間探索實(shí)驗(yàn)的平臺穿越次數(shù)和平臺象限游泳時間。 2．實(shí)驗(yàn)選用體重為18-22g健康昆明小鼠，雄性，132只，將實(shí)驗(yàn)小鼠隨機(jī)分為正常對照組（N group）、低氧預(yù)適應(yīng)組（HPC group）、假手術(shù)組（C group）、缺血再灌注組（O group）和HPC+O組，其中O組和HPC+O組又分為缺血再灌注1天、3天、7天和14天四個亞組，每組12只。HPC+O組于缺血再灌注前24h給予低氧預(yù)處理。O組和HPC+O組在相應(yīng)時間點(diǎn)處死取腦，C組在手術(shù)后24h處死取腦、HPC組在低氧預(yù)適應(yīng)后24h處死取腦，N組立即處死取腦。通過免疫熒光染色法對腦組織切片進(jìn)行檢測，于熒光顯微鏡下觀察海馬中CA1神經(jīng)細(xì)胞凋亡情況，并觀察MAP-2表達(dá)的分布，以及在海馬CA1區(qū)表達(dá)的動態(tài)變化。所有的實(shí)驗(yàn)數(shù)據(jù)均以均數(shù)±標(biāo)準(zhǔn)差（x s）表示，統(tǒng)計學(xué)檢驗(yàn)采用SPSS13.0軟件單因素方差分析（One WayANOVA），P0.05。 研究結(jié)果： 1．本實(shí)驗(yàn)成功復(fù)制低氧預(yù)適應(yīng)模型和全腦缺血再灌注模型。 2．水迷宮結(jié)果顯示：與N組相比，HPC組小鼠學(xué)習(xí)記憶能力明顯增加，差異有統(tǒng)計學(xué)意義，P0.05，而O組小鼠學(xué)習(xí)記憶能力明顯降低，P0.05；與O相比，HPC+O組小鼠學(xué)習(xí)記憶能力明顯改善，P 0.01。 3．熒光顯微鏡免疫熒光染色法檢測神經(jīng)細(xì)胞凋亡情況：缺血再灌注模型成功后，，凋亡神經(jīng)細(xì)胞在3d時表達(dá)最多；HPC+O3d組與N組、C組及HPC組相比，差異有統(tǒng)計學(xué)意義，P0.05。HPC+O1d組、HPC+O7d組、HPC+O14d組與N組、C組及HPC組相比，差異無統(tǒng)計學(xué)意義，P0.05。 4．熒光顯微鏡免疫熒光染色法測定MAP-2蛋白表達(dá)：缺血再灌注模型成功后，MAP-2陽性表達(dá)率增加在，7d時表達(dá)最多；與O組相比，HPC+O組MAP-2陽性表達(dá)明顯降低，統(tǒng)計學(xué)有顯著性差異，P0.05；N、C組及HPC組，MAP-2基本無表達(dá)。 結(jié)論： 1．低氧預(yù)適應(yīng)可以增加正常小鼠的學(xué)習(xí)記憶能力。 2．缺血再灌注腦損傷小鼠能出現(xiàn)不同程度的認(rèn)知功能障礙。 3．低氧預(yù)適應(yīng)能改善缺血再灌注腦損傷小鼠的學(xué)習(xí)記憶能力。 4．低氧預(yù)適應(yīng)能減少缺血性再灌注損傷小鼠的凋亡，起到神經(jīng)保護(hù)作用。 5．低氧預(yù)適應(yīng)能降低MAP-2陽性表達(dá)率，增加海馬區(qū)突觸可塑性，從而提高學(xué)習(xí)記憶的能力。 這對深入了解低氧預(yù)適應(yīng)對缺血再灌注性腦損傷學(xué)習(xí)記憶保護(hù)作用有重要意義，同時可為臨床上防治腦缺血/低氧后認(rèn)知功能損傷提供新策略和科學(xué)的實(shí)驗(yàn)依據(jù)。
[Abstract]:Purpose of study :

1 . To study the effects of hypoxic preconditioning on learning and memory of mice with cerebral ischemia - reperfusion injury .

2 . To study the effect of hypoxic preconditioning on neuronal apoptosis in CA1 region of hippocampus after ischemia - reperfusion .

3 . To study the effect of hypoxic preconditioning on MAP - 2 expression in CA1 region of hippocampus after ischemia - reperfusion .

Study method :

1 . Adult Kunming mice weighing 18 - 22 g were randomly divided into normal group ( Ngroup ) , sham operation group ( C group ) , ischemia / reperfusion group ( O group ) , hypoxic preconditioning group ( HPC group ) and hypoxic preconditioning + ischemia reperfusion group ( HPC + O group ) . 6 . The Morris water maze test was used to carry out the behavioral test , and the average latency and the space exploration experiment were recorded for the average latency and the platform quadrant swimming time .

2 . Experimental mice were randomly divided into three groups : normal control group ( N group ) , hypoxic preconditioning group ( HPC group ) , sham operation group ( C group ) , ischemia / reperfusion group ( O group ) and HPC + O group .

Results of the study :

1 . The hypoxic preconditioning model and global cerebral ischemia / reperfusion model were successfully replicated in the experiment .

2 . The water maze showed that the learning and memory ability of mice in HPC group was significantly higher than that in group N , P 0.05 , while the learning and memory ability of group O mice decreased significantly , P0.05 ;
Compared with O , the learning and memory ability of mice in HPC + O group was significantly improved , P 0.01 .

3 . Fluorescent microscopy immunofluorescence staining method was used to detect the apoptosis of nerve cells : after the ischemia / reperfusion model , the apoptotic nerve cells were expressed most at 3d ;
Compared with group N , group C and HPC group , HPC + O3d group had statistical significance , P0.05 . HPC + O1d group , HPC + O7d group , HPC + O14d group had no significant difference compared with group N , group C and HPC group , P0.05 .

4 . The expression of MAP - 2 protein was determined by fluorescence microscope immunofluorescence staining . After successful ischemia / reperfusion model , the expression of MAP - 2 was increased , and the expression of MAP - 2 was highest at 7d .
Compared with the O group , the positive expression of MAP - 2 in HPC + O group was significantly lower than that in O group ( P0.05 ) .
MAP - 2 was not expressed in group N , C and HPC .

Conclusion :

1 . hypoxic preconditioning can increase the learning and memory ability of normal mice .

2 . Different degrees of cognitive impairment can be found in the brain injury mice after ischemia / reperfusion .

3 . hypoxic preconditioning can improve the learning and memory ability of mice with cerebral ischemia - reperfusion injury .

4 . hypoxic preconditioning can reduce the apoptosis of ischemic reperfusion injury mice and play a role of neuroprotection .

5 . hypoxic preconditioning can decrease MAP - 2 positive expression rate , increase synaptic plasticity in hippocampus region , and improve learning and memory ability .

It is of great significance to understand the effect of hypoxic preconditioning on the learning and memory protection of ischemia - reperfusion brain injury , and it can provide new strategy and scientific experimental basis for the prevention and treatment of cognitive impairment after cerebral ischemia / hypoxia .

【學(xué)位授予單位】：泰山醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R743

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