缺糖氧再灌注后神经元氧化及抗氧化分子的变化及胞二磷胆碱的保护作用

发布时间：2018-03-22 18:52

本文选题：大鼠　切入点：海马神经元　出处：《南京医科大学》2008年硕士论文　论文类型：学位论文

【摘要】： 第一部分海马及皮层神经元体外原代培养及缺糖氧再灌注损伤易感性的比较目的:建立海马及皮层神经元原代培养、缺糖氧再灌注损伤模型,在此基础上分析海马、皮层神经元对缺糖氧再灌注损伤的易感性。方法:采用体外无血清培养法培养海马、皮层神经元,取8-10d的海马、皮层神经元,采用剥夺培养液氧糖法建立体外缺糖氧再灌注损伤模型,于缺糖氧再灌注3h、6h及24h时,比较观察两种神经元一氧化氮(NO)的释放量、谷胱甘肽(GSH)的含量及一氧化氮合酶(NOS)、谷胱甘肽过氧化物酶(GPx)的活性变化;于缺糖氧再灌注24h时,检测两种神经元乳酸脱氢酶(LDH)的释放量,并采用台盼蓝染色、Hochest33258神经元核染色观察两种神经元死亡、凋亡的改变。结果:缺糖氧再灌注后,海马神经元NOS的活性、NO的含量明显高于皮层神经元,而GPx的活性、GSH的含量低于皮层神经元(P＜0.05);缺糖氧再灌注后,海马神经元的死亡、凋亡率亦明显高于皮层神经元(P＜0.05)。结论:海马神经元相较于皮层神经元更易受缺糖氧再灌注的损伤,其可能与缺糖氧再灌注后海马神经元氧应激产物过多如NOS活性及NO释放量等的增高、抗氧化系统防御下降如GPx活性、GSH含量的下降有关。第二部分胞二磷胆碱对大鼠海马神经元缺糖氧再灌注损伤的保护作用目的观察胞二磷胆碱对大鼠海马神经元缺糖氧再灌注损伤的保护作用。方法建立海马神经元缺糖氧再灌注损伤模型,随机分组为正常对照组、缺糖氧再灌注组、胞二磷胆碱干预组(1、10、100umol/L),观察再灌注6、24h还原型谷胱甘肽含量以及谷胱甘肽过氧化物酶活性的改变;于再灌注24h检测海马神经元四甲基偶氮唑盐(MTT)代谢率,以及流式细胞术检测凋亡。结果与缺糖氧再灌注组相比,胞二磷胆碱于再灌注6h可明显提高GSH的含量及GPx的活性(P＜0.05);于再灌注24h可增高MTT代谢率,提高GPx的活性及减少海马神经元凋亡(P＜0.05)。结论胞二磷胆碱对海马神经元缺糖氧再灌损伤有明显的保护作用,其机制可能与改善神经元GSH含量、GPx活性及减少凋亡有关。
[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.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R363

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