板桥党参对激活GSK-3β诱导的AD模型大鼠认知功能障碍的保护作用及机制

发布时间：2018-03-27 16:23

本文选题：阿尔兹海默病　切入点：板桥党参　出处：《湖北民族学院》2017年硕士论文

【摘要】：目的:本课题主要研究板桥党参(Banqiao Codonopisis Pilosula,BCP)是否具有改善阿尔兹海默病(Alzheimer's disease,AD)模型大鼠认知功能障碍的作用,并通过检测大鼠海马组织Tau蛋白磷酸化水平及其相关激酶活性、神经发育等内容,探索其可能作用机制。方法:用不同剂量BCP水煎液对拟激活糖原合酶3β(Glycogen synthase 3β,GSK-3β)的AD模型大鼠进行灌胃处理。其中灌胃第9天进行Morris水迷宫训练,灌胃第13天进行侧脑室注射磷脂酰肌醇3-激酶特异性抑制剂渥曼青霉素(Wortmannin,WT)和蛋白激酶C特异性抑制剂GF-109203X(GFX),灌胃第14天进行水迷宫测试,检测各组大鼠空间认知功能。运用免疫印迹、免疫组织化学法检测大鼠海马Tau磷酸化水平变化情况及其与之密切相关的蛋白激酶GSK-3β活性变化,运用尼氏染色法观察大鼠海马区神经元形态变化以及神经细胞功能情况。结果:1.自主行为测试结果显示给大鼠灌服板桥党参水煎液后,三个治疗组均比未给药组大鼠自主活动次数减少,且随着时间推移自主活动次数逐渐减少。2.水迷宫测试表明治疗组能明显改善AD模型大鼠认知功能障碍。3.Western Blot结果显示AD模型组总的GSK-3β无变化,但是活性T216-GSK-3β比假手术组高(p0.05),而且p-Tau231、p-Tau262、p-Tau396、p-Tau404表达增加(p0.05)。板桥党参干预后Tau蛋白特殊位点磷酸化水平明显下调(p0.05)。4.免疫组织化学法检测脑海马组织Tau蛋白磷酸化修饰情况,结果显示:AD模型组海马CA3区Tau蛋白磷酸化程度高于对照组,而板桥党参干预组CA3区Tau蛋白磷酸化程度较模型组减低。5.尼氏染色结果显示:AD模型组CA1、CA3区神经元排列紊乱稀少,胞体呈空泡状,其尼氏小体染色比假手术组明显变浅,破碎状且变少,呈溶解状。板桥党参干预后神经元从形态和数量都有改善,神经元和尼氏小体数量、形态均明显恢复。结论:板桥党参对AD模型大鼠认知功能障碍具有一定的防治作用,其可能作用机制为降低T216-GSK-3β表达,抑制GSK-3β活性,最终下调Tau蛋白磷酸化水平,同时改善神经元形态和发育情况从而增强大鼠空间学习记忆能力。
[Abstract]:Objective: to study whether Banqiao Codonopisis Pilosula Codonopisis can improve cognitive dysfunction in rats with Alzheimer's disease (AD), and to detect the phosphorylation level of Tau protein and its kinase activity in hippocampus of rats. Methods: the AD rats were treated with different doses of BCP decoction to activate the glycogen synthase 3 尾 -glycogen synthase 3 尾 -GSK-3 尾, and the rats were given Morris water maze training on the 9th day. On the 13th day, intragastric injection of phosphatidylinositol 3-kinase specific inhibitor Wortmanninth WT) and protein kinase C-specific inhibitor GF-109203XFGFXX were performed on the 13th day. The water maze test was performed on the 14th day after intragastric administration. The spatial cognitive function of rats in each group was measured. The changes of Tau phosphorylation and protein kinase GSK-3 尾 activity in hippocampus of rats were detected by Western blot and immunohistochemistry. The morphological changes of neurons and the function of neurons in the hippocampus of rats were observed by using Nissl staining method. Results: 1. The results of autonomic behavior test showed that the rats were given the decoction of Panqiao Codonopsis pilosula (Codonopsis pilosula). The number of autonomous activities in the three treatment groups was lower than that in the control group. The water maze test showed that the treatment group could obviously improve the cognitive dysfunction of AD model rats. 3. Western Blot showed that the total GSK-3 尾 of AD model group had no change. However, the activity of T216-GSK-3 尾 was higher than that of sham-operated group, and the expression of p-Tau231 and p-Tau262, p-Tau396and p-Tau404 increased. The phosphorylation level of specific site of Tau protein was significantly down-regulated by Panqiao Codonopsis pilosula. 4. Immunohistochemical method was used to detect the phosphorylation modification of Tau protein in hippocampus tissue. The results showed that the phosphorylation degree of Tau protein in the CA3 region of hippocampus was higher in the control group than that in the control group, while the phosphorylation degree of Tau protein in the CA3 region of the control group was lower than that in the control group. The results of Nissl staining showed that the neurons in CA1 and CA3 region of the control group were poorly arranged. The cell bodies were vacuolated, and their Nissl bodies were obviously shallower, fragmented and less dissolved than those in the sham-operated group. After the intervention of Codonopsis pilosula, the number and morphology of neurons were improved, and the number of neurons and Nissl corpuscles were increased. Conclusion: Codonopsis pilosula has some preventive and therapeutic effects on cognitive dysfunction in AD model rats. The possible mechanism is to decrease the expression of T216-GSK-3 尾, inhibit the activity of GSK-3 尾, and eventually down-regulate the level of phosphorylation of Tau protein. At the same time, the morphology and development of neurons were improved to enhance the spatial learning and memory ability of rats.
【学位授予单位】：湖北民族学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5;R-332

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