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  本文選題：大鼠　切入點：海馬神經(jīng)元　出處：《南京醫(yī)科大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

【摘要】： 第一部分海馬及皮層神經(jīng)元體外原代培養(yǎng)及缺糖氧再灌注損傷易感性的比較 目的:建立海馬及皮層神經(jīng)元原代培養(yǎng)、缺糖氧再灌注損傷模型,在此基礎(chǔ)上分析海馬、皮層神經(jīng)元對缺糖氧再灌注損傷的易感性。方法:采用體外無血清培養(yǎng)法培養(yǎng)海馬、皮層神經(jīng)元,取8-10d的海馬、皮層神經(jīng)元,采用剝奪培養(yǎng)液氧糖法建立體外缺糖氧再灌注損傷模型,于缺糖氧再灌注3h、6h及24h時,比較觀察兩種神經(jīng)元一氧化氮(NO)的釋放量、谷胱甘肽(GSH)的含量及一氧化氮合酶(NOS)、谷胱甘肽過氧化物酶(GPx)的活性變化;于缺糖氧再灌注24h時,檢測兩種神經(jīng)元乳酸脫氫酶(LDH)的釋放量,并采用臺盼藍(lán)染色、Hochest33258神經(jīng)元核染色觀察兩種神經(jīng)元死亡、凋亡的改變。結(jié)果:缺糖氧再灌注后,海馬神經(jīng)元NOS的活性、NO的含量明顯高于皮層神經(jīng)元,而GPx的活性、GSH的含量低于皮層神經(jīng)元(P＜0.05);缺糖氧再灌注后,海馬神經(jīng)元的死亡、凋亡率亦明顯高于皮層神經(jīng)元(P＜0.05)。結(jié)論:海馬神經(jīng)元相較于皮層神經(jīng)元更易受缺糖氧再灌注的損傷,其可能與缺糖氧再灌注后海馬神經(jīng)元氧應(yīng)激產(chǎn)物過多如NOS活性及NO釋放量等的增高、抗氧化系統(tǒng)防御下降如GPx活性、GSH含量的下降有關(guān)。 第二部分胞二磷膽堿對大鼠海馬神經(jīng)元缺糖氧再灌注損傷的保護(hù)作用 目的觀察胞二磷膽堿對大鼠海馬神經(jīng)元缺糖氧再灌注損傷的保護(hù)作用。方法建立海馬神經(jīng)元缺糖氧再灌注損傷模型,隨機分組為正常對照組、缺糖氧再灌注組、胞二磷膽堿干預(yù)組(1、10、100umol/L),觀察再灌注6、24h還原型谷胱甘肽含量以及谷胱甘肽過氧化物酶活性的改變;于再灌注24h檢測海馬神經(jīng)元四甲基偶氮唑鹽(MTT)代謝率,以及流式細(xì)胞術(shù)檢測凋亡。結(jié)果與缺糖氧再灌注組相比,胞二磷膽堿于再灌注6h可明顯提高GSH的含量及GPx的活性(P＜0.05);于再灌注24h可增高M(jìn)TT代謝率,提高GPx的活性及減少海馬神經(jīng)元凋亡(P＜0.05)。結(jié)論胞二磷膽堿對海馬神經(jīng)元缺糖氧再灌損傷有明顯的保護(hù)作用,其機制可能與改善神經(jīng)元GSH含量、GPx活性及減少凋亡有關(guān)。
[Abstract]:Part I comparison of the susceptibility of primary culture of hippocampal and cortical neurons and reperfusion injury induced by glucose deficiency and oxygen deficiency in vitro. Objective: to establish the primary culture model of hippocampal and cortical neurons, and to analyze the susceptibility of hippocampal and cortical neurons to glucose deficiency and oxygen reperfusion injury. Methods: hippocampal hippocampus was cultured in vitro by serum-free culture. Cortical neurons were taken from hippocampal and cortical neurons for 8-10 days. The model of glucose deficiency reperfusion injury in vitro was established by deprivation of liquid oxygen sugar method. The release of nitric oxide (no) from the two neurons was observed at 3 h and 24 h after reperfusion. The content of glutathione (GSH) and the activity of nitric oxide synthase (NOS) and glutathione peroxidase (Glutathione peroxidase) were measured at 24 h after glucose deficiency reperfusion. The changes of neuronal death and apoptosis were observed by trypan blue staining and Hochest33258 neuronal nucleus staining. Results: the NOS activity of hippocampal neurons was significantly higher than that of cortical neurons. The activity of GPx was lower than that of cortical neurons (P < 0.05), and the death and apoptosis rate of hippocampal neurons were significantly higher than that of cortical neurons (P < 0.05). Conclusion: compared with cortical neurons, hippocampal neurons are more vulnerable to glycosylated oxygen reperfusion injury. It may be related to the increase of oxygen stress products such as NOS activity and no release in hippocampal neurons after glycosylated oxygen reperfusion, and the decrease of antioxidant system defense, such as the decrease of GPx activity. The second part of the protective effect of citicoline on rat hippocampal neuron injury induced by glucose deficiency and oxygen reperfusion. Objective to observe the protective effect of citicoline on rat hippocampal neuron injury induced by glucose deficiency and oxygen reperfusion. Methods the model of hippocampal neuron injury induced by glucose deficiency and oxygen deficiency reperfusion was established and divided into two groups randomly: normal control group and glucose deficient oxygen deficiency reperfusion group. The changes of glutathione content and glutathione peroxidase activity in hippocampal neurons were observed at 6h after reperfusion, and the metabolic rate of tetramethylazolazole (MTT) in hippocampal neurons was measured at 24 h after reperfusion. Results compared with the glucose deficient oxygen / oxygen reperfusion group, CDPC could significantly increase the content of GSH and the activity of GPx at 6 h after reperfusion (P < 0.05), and increase the metabolic rate of MTT at 24 h after reperfusion. Conclusion CDPC has an obvious protective effect on hippocampal neuronal injury induced by glucose deficiency and oxygen deprivation, and its mechanism may be related to the improvement of GSH activity and the decrease of apoptosis.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R363

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相關(guān)期刊論文 前1條

1 任鐵玲,胡前勝,傅洪軍,董勝璋;大鼠海馬神經(jīng)元無血清原代培養(yǎng)技術(shù)的建立[J];中國衛(wèi)生檢驗雜志;2004年02期

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本文編號：1649933