吡咯喹啉醌对神经细胞氧糖剥夺损伤的保护作用及机制研究

发布时间：2018-06-23 08:44

本文选题：吡咯喹啉醌 + 氧糖剥夺/复氧　；参考：《大连医科大学》2011年硕士论文

【摘要】：目的:脑缺血后的病理生理过程复杂,脑缺血再灌注损伤级联反应其中一环是细胞凋亡,抑制神经细胞凋亡有利于缺血性脑血管病的治疗和康复。近期国内外研究表明,吡咯喹啉醌(Pyrroloquinoline quinone ,PQQ)具有较强的抗氧化作用,可能通过抗氧化应激对脑缺血有保护作用。而氧化应激反应被认为是导致凋亡的最重要途径之一,因此本文研究吡咯喹啉醌(PQQ)是否具有抗凋亡作用及其可能机制。研究吡咯喹啉醌(PQQ)对体外培养小鼠神经母细胞瘤Neuro2A细胞株氧糖剥夺/复氧模型(OGD/R)的影响,在细胞水平探讨PQQ神经保护作用的可能机制。方法:利用不同浓度PQQ预处理Neuro2A细胞后,通过无氧密闭盒和无糖培养液,建立氧糖剥夺模型。取复氧后6h这一时间点,观察PQQ干预对Neuro2A细胞的细胞形态、存活率、凋亡率以及氧化应激检测指标活性氧(ROS)和还原型谷胱甘肽(GSH)的影响。细胞形态学采用倒置相差显微镜观察;细胞存活率采用四唑盐(MTT)法检测;细胞凋亡率采用荧光显微镜观察;ROS和GSH采用荧光分光光度计检测。结果:一.倒置相差显微镜观察细胞形态:OGD 2h复氧6h后Neuro2A细胞形态出现严重损伤,6.4μM和12.8μM的PQQ预处理有减轻神经细胞损伤的作用;二.MTT法检测细胞存活率:OGD 2h复氧6h后,0 12.8μM的PQQ预处理组细胞的存活率呈逐渐增高趋势(P0.05)。三.荧光显微镜下计数细胞凋亡率:OGD 2h复氧6h后,6.4μM、12.8μM的PQQ预处理组和OGD无保护组相比细胞凋亡数逐渐减少(P0.05)。四.PQQ预处理后细胞内ROS生成减少(P0.01),GSH水平增高(P0.05),与氧糖剥夺/复氧组相比有统计学差异。结论:一.PQQ对神经细胞氧糖剥夺损伤有一定的保护作用。二.PQQ对氧糖剥夺损伤的Neuro2A细胞的神经保护作用可能是通过减轻氧化应激反应,降低细胞凋亡率来实现的。
[Abstract]:Aim: the pathophysiological process after cerebral ischemia is complicated. One of the cascade reactions of cerebral ischemia reperfusion injury is apoptosis. Inhibition of neuronal apoptosis is beneficial to the treatment and rehabilitation of ischemic cerebrovascular disease. Recent studies at home and abroad have shown that pyrroloquinoline quinone (PQQ) has a strong antioxidant effect, which may have protective effect on cerebral ischemia through antioxidant stress. Oxidative stress reaction is considered to be one of the most important ways to induce apoptosis. Therefore, whether pyrroloquinoline quinone (PQQ) has anti-apoptotic effect and its possible mechanism is studied in this paper. To study the effect of pyrroloquinoline quinone (PQQ) on the oxygen deprivation / reoxygenation model (OGD / R) of mouse neuroblastoma cell line Neuro2A in vitro, and to explore the possible mechanism of PQQ neuroprotective effect at the cell level. Methods: Neuro2A cells were pretreated with different concentrations of PQQ. At 6 h after reoxygenation, the effects of PQQ on cell morphology, survival rate, apoptosis rate, reactive oxygen species (Ros) and reduced glutathione (GSH) in Neuro2A cells were observed. Cell morphology was observed by inverted phase contrast microscope, cell survival rate was detected by MTT assay, apoptosis rate was observed by fluorescence microscope and GSH was detected by fluorescence spectrophotometer. Results: 1. Observation of cell morphology by inverted phase contrast microscope (PQQ) after reoxygenation for 6 h after reoxygenation for 2 h at 1: 0GD could attenuate the injury of neuro2A cells by PQQ pretreatment of 6.4 渭 M and 12.8 渭 M, respectively. 2. MTT assay was used to detect the survival rate of cells in the PQQ pretreatment group with 12.8 渭 M of 0 渭 M after reoxygenation for 2 h and 6 h after reoxygenation for 2 h (P0.05). III. The apoptosis rate was counted under fluorescence microscope. The apoptosis rate of PQQ pretreatment group (6.4 渭 M) 12.8 渭 M after reoxygenation for 6 h after reoxygenation for 2 h was gradually decreased compared with that in OGD unprotected group (P0.05). 4. After pretreatment with PQQ, Ros production decreased (P0.01) and GSH level increased (P0.05), which was significantly different from that of glucose deprivation / reoxygenation group. Conclusion: 1. PQQ has protective effect on neuronal oxygen and glucose deprivation injury. 2. The neuroprotective effect of PQQ on neuro2A cells injured by oxygen glucose deprivation may be achieved by reducing oxidative stress response and apoptosis rate.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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