自主跑轮运动通过抑制NF-κB通路改善阿尔茨海默病模型小鼠学习记忆能力

发布时间：2018-04-25 08:17

本文选题：阿尔茨海默病 + 自主跑轮运动　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:观察自主跑轮运动对阿尔茨海默病(Alzheimer’s disease,AD)模型小鼠学习记忆能力和海马内炎性细胞因子表达的影响,探讨自主跑轮运动改善AD学习记忆能力和神经炎症的机制,为神经退行性疾病的防治提供新思路。方法:选取体重为30~35g的三月龄雄性昆明小鼠45只,由山西医科大学实验动物中心提供,随机分成三组:(1)DMSO对照组;(2)AD安静组;(3)AD运动组。每组15只。用3.5%水合氯醛腹腔注射麻醉小鼠,根据小鼠脑图谱,采用脑立体定位仪,对DMSO对照组进行侧脑室注射DMSO,AD安静组和AD运动组进行侧脑室注射Aβ1-42寡聚体。侧脑室注射1周后,AD运动组小鼠进行为期6周的自主跑轮运动,DMSO对照组和AD安静组进行正常饲养,不运动。6周自主跑轮运动结束后,采用Morris水迷宫实验,评估自主跑轮运动对AD模型小鼠学习记忆能力的影响。采用免疫荧光染色方法检测小鼠海马内肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、白介素10(Interleukin 10,IL-10)、核转录因子κB(Nuclear factor Kappa B,NF-κB)的阳性表达率变化。采用蛋白印迹方法(Western blot)检测小鼠海马内上述指标的蛋白表达变化。结果:Morris水迷宫实验结果显示,6周自主跑轮运动结束后,AD运动组的逃避潜伏期(12.10±1.26)显著低于AD安静组(27.75±3.88)(P㩳0.01),提示6周的自主跑轮运动有效改善了AD模型小鼠的学习记忆能力。免疫荧光染色结果显示,NF-κB、TNF-α和IL-10在DMSO对照组、AD安静组和AD运动组海马CA1区和CA3区均有表达,且各自在两区表达趋势一致。在海马CA1区和CA3区,NF-κB和促炎细胞因子TNF-α在AD安静组的阳性表达率均明显高于其他两组(P㩳0.05);而IL-10在AD安静组的阳性表达率明显低于AD运动组和DMSO对照组(P㩳0.05);而上述指标在AD运动组和DMSO对照组的阳性表达率无明显差异(P㧐0.05)。Western blot结果显示,在小鼠海马中,NF-κB和促炎细胞因子TNF-α在AD安静组的蛋白表达量最高,与其余两组均具有统计学差异(P㩳0.05);IL-10在AD安静组的蛋白表达量最低,在AD运动组和DMSO对照组表达较高,且后两者表达量较为接近,都与AD安静组存在统计学差异(P㩳0.05)。结论:(1)自主跑轮运动有效改善了AD模型小鼠的学习记忆能力。(2)自主跑轮运动明显下调了AD模型小鼠海马内促炎细胞因子TNF-α的释放;上调了抗炎细胞因子IL-10的表达。(3)NF-κB通路的抑制在自主跑轮运动改善AD模型小鼠学习记忆能力方面起到关键作用。自主跑轮运动能够显著抑制过度活化的NF-κB、TNF-α级联炎症反应,并通过调控海马内炎症反应失衡状态,改善AD模型小鼠学习记忆能力。
[Abstract]:Objective: to observe the effects of self-running wheel exercise on learning and memory ability and the expression of inflammatory cytokines in hippocampus of Alzheimer's disease (AD) mice, and to explore the mechanism of self-running wheel exercise in improving AD learning and memory ability and neuroinflammation. To provide a new idea for the prevention and treatment of neurodegenerative diseases. Methods: a total of 45 male Kunming mice weighing 30 ~ 35g were randomly divided into three groups: the control group was divided into three groups. There were 15 rats in each group. Anesthetized mice were injected intraperitoneally with 3.5% chloral hydrate. According to the brain map of mice, A 尾 1-42 oligodeoxynucleotides were injected into the lateral ventricle of DMSO control group by intracerebroventricular injection of DMSO-AD quiescent group and AD exercise group by intracerebroventricular injection of A 尾 1-42 oligomer. One week after intracerebroventricular injection, the mice in AD exercise group were fed with DMSO control group and AD quiet group for 6 weeks, and the Morris water maze test was used after 6 weeks of exercise. To evaluate the effect of self-running wheel exercise on learning and memory ability of AD model mice. The positive expression of tumor necrosis factor- 伪 (Tumor necrosis factor- 伪), interleukin-10 (10(Interleukin 10) IL-10 and nuclear transcription factor 魏 B(Nuclear factor Kappa (NF- 魏 B) in the hippocampus of mice were detected by immunofluorescence staining. Western blot was used to detect the protein expression of the above indexes in the hippocampus of mice. Results the results of the water maze test showed that the escape latency of AD group was significantly lower than that of AD group (27.75 卤3.88) after 6-week self-running. It suggested that 6-week self-running exercise could effectively improve the learning and memory ability of AD model mice. The results of immunofluorescence staining showed that TNF- 伪 and IL-10 were expressed in the CA1 and CA3 regions of hippocampus in AD quiet group and AD exercise group in DMSO control group, and the expression trend was the same in the two regions. The positive expression rates of NF- 魏 B and TNF- 伪 in hippocampal CA1 and CA3 regions were significantly higher in the quiet AD group than in the other two groups, while the positive expression rate of IL-10 in the AD quiet group was significantly lower than that in the AD exercise group and the DMSO control group. There was no significant difference in the positive expression rate between AD exercise group and DMSO control group. The results of Western blot showed that there was no significant difference between AD exercise group and DMSO control group. In the hippocampus of mice, the expression of NF- 魏 B and TNF- 伪 was the highest in the quiet group of AD, and there was a statistical difference between the two groups. The expression of IL-10 was the lowest in the quiet group of AD, and was higher in the exercise group and DMSO control group. The expression of the latter two groups was similar to that of the quiet group, and there was a statistical difference between the two groups. Conclusion: (1) the ability of learning and memory of AD model mice was improved effectively by self-running wheel exercise.) the release of pro-inflammatory cytokine TNF- 伪 in hippocampus of AD model mice was significantly down-regulated by autonomic wheel running. The inhibition of NF- 魏 B pathway up-regulated the expression of anti-inflammatory cytokine IL-10 played a key role in improving the learning and memory ability of AD model mice. The autonomic wheel running could significantly inhibit the over-activated inflammatory reaction of NF- 魏 B- TNF- 伪 cascade, and improve the learning and memory ability of AD model mice by regulating the imbalance of inflammatory response in hippocampus.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.16;R-332

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