OSAS模型大鼠海马区DDHD2和ERC2表达及突触变化的实验研究

发布时间：2018-01-18 12:34

本文关键词：OSAS模型大鼠海马区DDHD2和ERC2表达及突触变化的实验研究　出处：《遵义医学院》2017年硕士论文　论文类型：学位论文

【摘要】：目的:监测阻塞性睡眠呼吸暂停综合征(Obstructive sleep apnea syndrome,OSAS)模型大鼠空间学习记忆能力变化。检测实验大鼠海马区DDHD2及ERC2蛋白的表达变化,并观察突触前活动区超微结构变化。方法:22只雄性SD大鼠随机分为正常对照组与OSAS模型组,每组11只。通过设置慢性间歇性缺氧(Chronic intermittent hypoxia,CIH)环境,模拟OSAS过程,建立大鼠实验模型。用Morris水迷宫(Morris water maze,MWM)观察实验大鼠空间学习与记忆能力变化;使用蛋白质印迹法(Western Blot,WB)观察实验大鼠海马区DDHD2及ERC2蛋白的表达变化;并通过免疫组织化学(immunohistochemistry,I HC)法观察DDHD2及ERC2在海马CA1区神经细胞的表达定位。最后在透射电镜下,观察突触前活动区超微结构的变化。结果:OSAS模型建立评估:与常氧环境比较,低氧环境中的OSAS实验大鼠出现了活动减少、睡眠增多、呼吸频率加快及抬头呼吸等缺氧表现。同时监测低氧环境中大鼠尾动脉血氧饱和度(Sp O2)波动在69%至82%之间,常氧环境中Sp O2波动在96%至100%之间。MWM结果:定位航行试验中两组实验大鼠的逃避潜伏期随着每天重复训练的增加而不断缩短。正常对照组为(第1天39.4±11.0,第2天32.4±7.9,第3天17.4±5.9,第4天11.4±4.8,第5天8.0±3.1);OSAS模型组为(第1天46.8±12.4,第2天43.6±10.7,第3天32.3±9.5,第4天26.6±8.1,第5天22.1±7.1)。不同天数模型组逃避潜伏期均较正常组延长,两组差异具有统计学意义(P0.05)。空间探索试验:实验大鼠穿越原目标平台的次数为(正常对照组5.3±2.1,OSAS模型组3.1±1.7),结果表明模型组大鼠穿越原目标平台的次数较正常组减少,差异具有统计学意义(P0.05);在目标象限的时间百分比为(正常对照组31.5±5.6,OSAS模型组24.2±7.1),结果显示模型组大鼠在目标象限的时间百分比较正常组降低,差异具有统计学意义(P0.05)。Western Blot结果:OSAS模型大鼠海马区DDHD2蛋白表达的灰度值较正常组增高(正常组1.03±0.39,OSAS模型组1.47±0.37);模型组ERC2蛋白表达的灰度值较正常组降低(正常组0.50±0.20,OSAS模型组0.29±0.14),差异均具有统计学意义(P0.05)。免疫组化法观察到DDHD2在两组实验大鼠海马CA1区的神经细胞胞浆及胞核内表达,ERC2在实验大鼠海马CA1区神经细胞胞浆内表达。透射电镜结果:模型组大鼠海马CA1区突触前活动区长度减少(正常组0.30±0.05,OSAS模型组0.21±0.06),与正常组比较有显著差异(P0.01)。结论:CIH模拟OSAS所建立的实验动物模型,出现了学习、记忆能力等认知功能障碍及突触前超微结构变化,其可能与伴随的DDHD2过表达和ERC2的表达降低有关。
[Abstract]:Objective: to monitor obstructive sleep apnea syndrome in obstructive sleep apnea syndrome (OSAS). The changes of spatial learning and memory ability and the expression of DDHD2 and ERC2 in hippocampal region of experimental rats were detected. Methods 22 male Sprague-Dawley rats were randomly divided into normal control group and OSAS model group. 11 rats in each group were used to simulate the OSAS process by setting up chronic intermittent hypoxia (Chronic intermittent hypoxiaicus) environment. Morris water maze was used to observe the changes of spatial learning and memory ability of experimental rats. The expression of DDHD2 and ERC2 in hippocampal region of experimental rats was observed by Western blotblotWB (Western blotting). Immunohistochemical method was used. The expression and localization of DDHD2 and ERC2 in hippocampal CA1 neurons were observed by I-HCI method. Finally, transmission electron microscopy was used. Results: compared with normoxic environment, OSAS rats in hypoxic environment showed decreased activity and increased sleep. The oxygen saturation of caudal artery in hypoxic environment was monitored from 69% to 82%. SPO _ 2 fluctuates between 96% and 100% in normoxic environment. MWM results:. The escape latency of the two groups decreased with the increase of daily repeated training. On day 1, 39.4 卤11.0. 32.4 卤7.9 on the second day, 17.4 卤5.9 on the third day, 11.4 卤4.8 on the fourth day, 8.0 卤3.1 on the fifth day. The OSAS model group was 46.8 卤12.4 on the first day, 43.6 卤10.7 on the second day, 32.3 卤9.5 on the third day and 26.6 卤8.1 on the fourth day. On the 5th day, the escape latency of the model group was longer than that of the normal group. The difference between the two groups was statistically significant (P 0.05). Space exploration test: the number of experimental rats crossing the original target platform was 5.3 卤2.1 (normal control group). The results showed that the number of crossing the original target platform in the model group was less than that in the normal group, and the difference was statistically significant (P 0.05). The percentage of time in the target quadrant was 24.2 卤7.1 in the normal control group (31.5 卤5.6). The results showed that the percentage of time in the target quadrant of the model group was lower than that of the normal group. The difference was statistically significant (P 0.05). Western Blot results showed that the gray level of DDHD2 protein expression in the hippocampus of the rats with 1% OSAS was higher than that of the normal group (2). Normal group 1.03 卤0.39. OSAS model group (1.47 卤0.37); The expression of ERC2 protein in the model group was significantly lower than that in the normal group (0.50 卤0.20 卤0.29 卤0.14). The difference was statistically significant (P 0.05). The expression of DDHD2 in the cytoplasm and nucleus of hippocampal CA1 of the two groups was observed by immunohistochemical method. The results of transmission electron microscope showed that the length of presynaptic active area in CA1 region of hippocampus of model group was decreased (0. 30 卤0. 05). There was significant difference between OSAS model group and normal group (0.21 卤0.06). Conclusion the experimental animal model established by OSAS simulated by OSAS has appeared learning. Cognitive impairment such as memory ability and ultrastructural changes in presynaptic process may be related to the overexpression of DDHD2 and the decrease of ERC2 expression.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R766;R-332

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