β-胡萝卜素对OSAS大鼠学习记忆及海马区Caspase-3、p-tau表达变化的影响

发布时间：2018-04-25 20:23

本文选题：睡眠呼吸暂停 + 间歇缺氧　；参考：《遵义医学院》2017年硕士论文

【摘要】：目的:通过模拟阻塞性睡眠呼吸暂停综合征(OSAS)的病理特征即慢性间歇性缺氧(CIH)建立OSAS动物模型,观察β-胡萝卜素(β-C)对OSAS大鼠学习记忆及其海马区caspase-3、p-tau表达变化的影响。方法:将24只成年SD大鼠随机分为正常组、OSAS模型组和β-C干预组,每组各8只。采用低氧舱建立OSAS大鼠动物模型。β-C溶于蒸馏水中于造模前灌胃给药,正常组不做任何处理。造模完成后,采用Morris水迷宫检测实验大鼠的学习和记忆能力。HE染色观察大鼠海马组织的病理学改变。免疫组化染色检测磷酸化tau蛋白(p-tau)在大鼠海马组织中的表达部位。蛋白免疫印迹(Western Blot)检测海马组织中caspase-3、p-tau的蛋白表达变化。结果:OSAS模型判定结果:大鼠鼠尾动脉血氧饱和度平均值在CIH缺氧期为(69.72±1.64)%,相比于复氧期(95.38±1.53)%明显降低,且下降幅度4%,过程中大鼠出现抬头呼吸、憋醒以及腹式呼吸加深加快等症状,符合OSAS模型判定标准。Morris水迷宫行为学测试结果:在定位航行实验中,逃避潜伏期时间第1天正常组为(36.06±7.61)秒,模型组为(71.38±11.63)秒,干预组为(46.78±11.49)秒;第2天正常组为(26.44±6.46)秒,模型组为(63.12±9.06)秒,干预组为(37.16±5.66)秒;第3天正常组为(23.86±6.08)秒,模型组为(57.65±7.33)秒,干预组为(34.76±8.57)秒;第4天正常组为(20.25±4.81)秒,模型组为(51.89±7.21)秒,干预组为(30.40±6.85)秒;第5天正常组为(17.52±4.33)秒,模型组为(51.57±7.32)秒,干预组为(25.75±5.46)秒;与正常组比较,模型组用时明显增长(P0.05),而干预组相比于模型组则用时减少(P0.05)。在空间探索实验中,大鼠穿越平台的次数正常组为(7.03±1.49)次,模型组为(3.06±1.07)次,干预组为(5.13±1.36)次,模型组与正常组、干预组相比,穿越平台的次数明显减少,其结果均具有统计学意义(P0.05)。HE染色结果:与正常组大鼠相比,模型组和干预组的大鼠海马区神经细胞排列稀疏、紊乱,细胞形态不规则,细胞核固缩、裂解,细胞变性坏死,干预组较模型组程度轻。免疫组化结果:OSAS大鼠海马区p-tau阳性染色细胞呈棕黄色,主要表达于神经细胞包膜,细胞核和细胞质也有部分表达。Western Blot检测结果:模型组大鼠海马区caspase-3、p-tau的蛋白表达明显高于正常组和β-C干预组,差异具有统计学意义(P0.05)。结论:β-C可减轻OSAS所致的学习记忆能力受损,其机制可能与其抑制caspase-3、p-tau的蛋白表达有关。
[Abstract]:Objective: to establish OSAS animal model by simulating the pathological characteristics of obstructive sleep apnea syndrome (OSAS), and to observe the effect of 尾 -carotene (尾 -C-) on learning and memory and the expression of caspase-3 ~ (3) -p-tau in hippocampus of OSAS rats. Methods: Twenty-four adult SD rats were randomly divided into normal group (n = 8) and 尾 -C intervention group (n = 8). OSAS rat model was established in hypoxic chamber. 尾 -C dissolved in distilled water was administered intragastrically before modeling, but no treatment was given in normal group. Morris water maze was used to detect the learning and memory ability of the experimental rats. He staining was used to observe the histopathological changes of the hippocampal tissues of the rats. Immunohistochemical staining was used to detect the expression of phosphorylated tau protein p-tau) in rat hippocampus. Western blot was used to detect the expression of caspase-3 and tau in hippocampal tissue. Results: the average oxygen saturation of caudal artery of rats was 69.72 卤1.64 in the anoxic phase of CIH, which was significantly lower than that in the reoxygenation period (95.38 卤1.53%), and the decrease was 4%. During the process, the rats appeared the symptoms of rising breathing, suffocating waking and accelerating abdominal respiration. According to the criteria of OSAS model determination. Morris water maze behavior test results: in the localization navigation experiment, the escape latency was 36.06 卤7.61) seconds in the normal group, 71.38 卤11.63) seconds in the model group, 46.78 卤11.49) seconds in the intervention group and 26.44 卤6.46) seconds in the normal group on the second day. It was 63.12 卤9.06 seconds in the model group, 37.16 卤5.66 seconds in the intervention group, 23.86 卤6.08 seconds in the normal group on the 3rd day, 57.65 卤7.33 seconds in the model group, 34.76 卤8.57 seconds in the intervention group, 20.25 卤4.81) seconds in the normal group on the 4th day, 30.40 卤6.85 seconds in the intervention group and 17.52 卤4.33) seconds in the normal group on the 5th day. Compared with the normal group, the time used in the model group was significantly higher than that in the model group (P 0.05), while the time used in the intervention group was less than that in the model group. The model group was 51.57 卤7.32 seconds and the intervention group was 25.75 卤5.46 seconds. In the space exploration experiment, the number of rats crossing the platform was 7.03 卤1.49 times in the normal group, 3.06 卤1.07 times in the model group and 5.13 卤1.36 times in the intervention group. Compared with the normal group, the number of crossing the platform in the model group was significantly lower than that in the intervention group. Compared with the normal group, the hippocampal neurons in the model group and the intervention group were sparsely arranged, disordered, irregular in shape, pyknosis, cleavage, degeneration and necrosis. The intervention group was lighter than the model group. Immunohistochemical results showed that the p-tau positive staining cells in the hippocampal area of the rats were brown and mainly expressed in the capsule of nerve cells. The expression of caspase-3 p-tau in hippocampus of the model group was significantly higher than that of the normal group and 尾 -C intervention group, and the difference was statistically significant (P 0.05). Conclusion: 尾 -C can attenuate the impairment of learning and memory induced by OSAS, and its mechanism may be related to its inhibition of the expression of caspase-3 p-tau protein.
【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

【相似文献】