DRP-2在苯丙胺致小鼠脑损伤中的变化和机制

发布时间：2018-03-15 20:03

本文选题：苯丙胺　切入点：脑　出处：《暨南大学》2010年硕士论文　论文类型：学位论文

【摘要】： 目的：探讨二氢嘧啶酶相关蛋白2(DRP-2)在苯丙胺致小鼠脑损伤中的变化和机制。方法：将60只健康雄性C57BL／6小鼠随机分为：正常对照组、生理盐水组和苯丙胺组。苯丙胺组又分为1 d、7d、14d和28 d四个组,腹腔注射苯丙胺2 mg/kg/d。生理盐水组仅设28 d组,等量生理盐水注射。正常组不予任何处理。建模期间进行小鼠自主行为活动测试,电脑记录运动总路程、运动总时间、平均速度、最大速度、慢速运动时间、慢速运动时间／总时间、快速运动时间和快速运动时间／总时间等指标,检测动物的自主活动度。动物模型建立完成后,按照实验所需方法取材,采用银染(Nauta)法、免疫组织化学法及Western blotting法等观察苯丙胺对小鼠脑的毒性损害作用以及对DRP-2及其上游AKT/GSK-3β信号通路的影响。结果： 1.自主行为学检测：用药前各组小鼠的最大速度、平均速度、运动总路程、快速运动时间／总时间均没有显著差异(P0.05)。用药后,苯丙胺7 d、14 d、28 d组的运动总路程、平均速度、快速运动时间／总时间／最大运动速度比正常对照组和生理盐水组增加(P0.05),而慢速运动时间／总时间在各组间没有显著差异(P0.05)。 2.溃变神经纤维及其终末的银染结果：Nauta法染色显示苯丙胺14 d和28 d组纹状体内可见变性神经纤维呈黑色,正常组和生理盐水组均未发现变性的神经纤维。 3.多巴胺(DA)能神经元及其纤维的变化：苯丙胺28 d组小鼠黑质致密部的DA能神经元数比其它各组均减少(P0.05)。苯丙胺1d组和生理盐水组苍白球区DA神经纤维密度比正常组增加(P0.05),但随着用药时间的延长,苯丙胺14d和28 d组苍白球区的DA神经纤维密度比正常对照组和生理盐水组降低(P0.05)。 4.DRP-2及其蛋白激酶B／糖原合成激酶3p(AKT/GSK-3β)信号通路的变化：苯丙胺组纹状体内DRP-2的表达较生理盐水组和正常组明显减少(P0.05),而P-DRP-2的表达苯丙胺1d组与正常组无明显变化(P0.05),苯丙胺7d、14d组和生理盐水组较正常组增加(P0.05)。苯丙胺28d组AKT的表达较正常组和生理盐水组减少(P0.05)。苯丙胺7d、14d和28d组GSK-3β的表达比正常组增多(P0.05)。结论： 1.苯丙胺可引起小鼠自主活动性增加。 2.长期使用苯丙胺可导致起小鼠纹状体神经纤维变性。 3.长期使用苯丙胺可导致起小鼠黑质致密部的DA能神经元数目减少,以及苍白球区DA神经纤维密度的降低。 4.使用苯丙胺可引起小鼠纹状体P-DRP-2的增加以及DRP-2的减少,其机理与抑制AKT/GSK-3β信号通路的有密切关系。
[Abstract]:Objective:. To investigate the changes and mechanism of dihydropyrimidinase-associated protein 2DRP-2 in brain injury induced by amphetamine in mice. Methods:. Sixty healthy male C57BL / 6 mice were randomly divided into normal control group, saline group and amphetamine group. The normal group did not receive any treatment. During the modeling period, the mice were tested for autonomous behavior, and the total distance, time, average speed, maximum speed, slow motion time were recorded by computer. The indexes of slow motion time / total time, fast motion time and fast motion time / total time were used to detect the autonomous activity of animals. After the establishment of the animal model, the materials were obtained according to the method needed in the experiment, and the silver staining Nauta method was used. Immunohistochemical method and Western blotting method were used to observe the toxic effects of amphetamine on the brain of mice and the effects on DRP-2 and its upstream AKT/GSK-3 尾 signaling pathway. Results:. 1. Detection of autonomic behavior: there was no significant difference in maximal velocity, average speed, total distance, time / total time of rapid exercise in each group before treatment. After treatment, the total distance and average speed of the rats in the group of amphetamine 7 days and 14 days and 28 days after treatment were not significantly different. The rapid exercise time / total time / maximum exercise velocity increased P0.05 compared with normal control group and normal saline group, but there was no significant difference in slow exercise time / total time between groups. 2. The results of silver staining of degenerated nerve fibers and their terminals showed that denatured nerve fibers were black in striatum of 14 d and 28 d groups of amphetamine, but no denatured nerve fibers were found in normal group and normal saline group. 3. Changes of dopaminergic neurons and their fibers: the number of DA neurons in the substantia nigra of the rats in the 28 d group was lower than that in the other groups (P 0.05), and the density ratio of DA fibers in the globus pallidus was positive in the 1 d group and the saline group. In the normal group, P0.05 was increased, but with the prolongation of the medication time, The density of DA nerve fibers in the globus pallidus of 14 d and 28 d group was lower than that of normal control group and normal saline group. 4. The changes of signal pathway of DRP-2 and its protein kinase B / glycogen synthesis kinase 3pAK / GSK-3 尾: the expression of DRP-2 in striatum of amphetamine group was significantly lower than that of normal saline group and normal group, but the expression of P-DRP-2 was not significantly changed in phenylalanine 1 day group and normal group (P0.05). Compared with the control group, the expression of AKT in the amphetamine 28d group was lower than that in the normal saline group (P 0.05). The expression of GSK-3 尾 in the phenylalanine group was significantly higher than that in the control group on day 14 and 28. The expression of GSK-3 尾 in the control group was significantly higher than that in the control group (P 0.05). Conclusion:. 1. Amphetamine induced increased autonomous activity in mice. 2. Long-term use of amphetamine can lead to degeneration of striatum nerve fibers in mice. 3. The number of DA neurons in the substantia nigra and the density of DA nerve fibers in the ganglia pallidus were decreased after long-term use of amphetamine. 4. The increase of P-DRP-2 and the decrease of DRP-2 in the striatum of mice induced by amphetamine were closely related to the inhibition of AKT/GSK-3 尾 signaling pathway.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363

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