本文選題：老年性癡呆　切入點(diǎn)：學(xué)習(xí)記憶　出處：《華東師范大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

【摘要】： [目的] 運(yùn)用生化、組織、行為學(xué)方法評價(jià)D-半乳糖引發(fā)老年性癡呆(Alzheimer’s Disease，，AD)小鼠模型，測定海馬腦區(qū)甘丙肽及其受體表達(dá)的變化；探討甘丙肽在AD發(fā)生過程中可能的作用及其機(jī)制。 [方法] 選用20g左右的BL/C57小鼠50只，分為模型組和對照組，各25只。模型組腹腔注射D-半乳糖150mg/kg，每天一次，連續(xù)90d，制備老年癡呆動(dòng)物模型。對照組注射等量的0.9％的生理鹽水。 通過Y-型迷宮比較兩組小鼠的學(xué)習(xí)記憶能力，第一天檢測小鼠學(xué)習(xí)能力，間隔24小時(shí)后檢測小鼠記憶能力。 行為學(xué)實(shí)驗(yàn)結(jié)束后，每組取10只小鼠，測定蛋白質(zhì)含量和AChE、ChAT、MAO、MDA、GSH-PX、SOD、Na~+K~+-ATP酶，按試劑盒說明書所述步驟操作。 各組小鼠腹腔注射戊巴比妥鈉60mg/kg麻醉，用多聚甲醛進(jìn)行心臟灌流，，取腦感覺運(yùn)動(dòng)區(qū)皮層，置于戊二醛中，透射電鏡觀察。 熒光顯微鏡配合免疫組織化學(xué)觀察小鼠腦的矢狀冷凍切片海馬區(qū)域甘丙肽的表達(dá)，熒光顯微鏡檢測照相。 Realtime-PCR檢測小鼠腦區(qū)前額葉、中腦導(dǎo)水管周圍灰質(zhì)中甘丙肽及其受體1、2、3表達(dá)狀況。 [結(jié)果] (1)行為學(xué)檢測結(jié)果說明模型小鼠的學(xué)習(xí)記憶能力明顯低于對照小鼠。 (2)生化指標(biāo)檢測出模型組AChE、ChAT、GSH-PX、SOD、Na~+K~+-ATP酶活性明顯降低，MAO活性上升，MDA含量明顯上升。 (3)電鏡病理切片顯示模型組的細(xì)胞核明顯萎縮；線粒體嵴消失，或產(chǎn)生空泡。對照組細(xì)胞核正常，線粒體嵴清晰。 (4)免疫熒光檢測海馬區(qū)域甘丙肽含量表達(dá)差異，模型組甘丙肽表達(dá)明顯高于對照組。 (5) Realtime-PCR檢測出模型組前額葉、中腦導(dǎo)水管周圍灰質(zhì)區(qū)域甘丙肽及其受體1、2、3的mRNA含量高于對照組。 [結(jié)論] (1)模型建立比較成功，為實(shí)驗(yàn)提供可靠的前提。 (2)老年癡呆小鼠氧化還原酶SOD、GSH-PX、AChE、ChAT、Na~+-K~+-ATP酶活性下降，MDA含量上升、MAO活性上升，說明體內(nèi)氧自由基含量明顯升高，符合衰老的特征。 (3)癡呆小鼠的腦細(xì)胞發(fā)生了病理變化：細(xì)胞核萎縮，線粒體破裂或呈空泡。 (4)甘丙肽在老年癡呆小鼠海馬中呈陽性增加表達(dá)效應(yīng)。 (5)前額葉、中腦導(dǎo)水管周圍灰質(zhì)腦區(qū)甘丙肽受體出現(xiàn)高表達(dá)。
[Abstract]:[purpose]. Methods of biochemistry, tissue and behavior were used to evaluate the model of Alzheimer's dementia induced by D-galactose-induced Alzheimer's Disease (AD), to determine the changes of glycine and its receptor expression in hippocampus, and to explore the possible role and mechanism of galanin in the pathogenesis of AD. [methods]. Fifty BL/C57 mice with 20 g or so were divided into two groups: model group and control group. The model group was injected intraperitoneally with D-galactose 150 mg / kg, once a day for 90 days, and the control group was injected with the same amount of 0.9% normal saline. The learning and memory abilities of the two groups were compared by Y- maze. The learning ability of the mice was measured on the first day and the memory ability of the mice was measured after 24 hours interval. At the end of behavioral experiment, 10 mice in each group were selected. The protein content and the protein content in each group were determined, and the activity of GSH-PXX SODN Na ~ K ~ -ATPase was determined according to the procedure described in the specification of the kit. The mice were anesthetized with 60 mg / kg pentobarbital sodium intraperitoneally and perfused with paraformaldehyde. The cortex of sensorimotor area was taken and placed in glutaraldehyde and observed by transmission electron microscope. The expression of galanin in hippocampus of sagittal frozen section of mouse brain was observed by fluorescence microscope and immunohistochemistry. Realtime-PCR was used to detect the expression of glycidyl peptide and its receptor 1 / 2 in the prefrontal lobe and periaqueductal gray of mice. [results]. The results of behavioral examination showed that the learning and memory ability of the model mice was significantly lower than that of the control mice. In the model group, the activity of GSH-PXX SODN Na ~ K ~ -ATPase decreased obviously and the content of MDA increased obviously. Electron microscopic pathological sections showed that the nuclei of the model group were obviously atrophied, the mitochondrial crest disappeared or vacuoles were produced, and the nuclei of the control group were normal and the mitochondrial crest was clear. (4) Immunofluorescence assay showed that the level of galanin in the model group was significantly higher than that in the control group. (5) the content of mRNA in prefrontal lobe and periaqueductal gray matter area of model group was higher than that in control group by Realtime-PCR. [conclusion]. 1) the establishment of the model is successful and provides a reliable premise for the experiment. (2) in senile dementia mice, the activity of redox enzyme SODH-PXH- AChEATE (Na ~ -K ~ -ATPase) decreased and the activity of Mao increased, indicating that the content of oxygen free radicals in the body was obviously increased, which was consistent with the characteristics of senility. 3) pathological changes of brain cells in dementia mice: nucleus atrophy, mitochondria rupture or vacuole. 4) the expression of glycidyl in hippocampus of senile dementia mice was increased. The prefrontal lobe and periaqueductal gray area of the midbrain showed high expression of glycine receptor.
【學(xué)位授予單位】：華東師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R-332

【引證文獻(xiàn)】

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1 榮曼;pcDNA3.1-PDGF β -GAL在Neuro-2a細(xì)胞的表達(dá)及其對細(xì)胞增殖和周期的影響[D];山西醫(yī)科大學(xué);2013年

本文編號(hào)：1593237