侧脑室注射GDNF对甲基苯丙胺依赖鼠伏隔核DRD2、DAT及其mRNA变化的影响

发布时间：2018-04-02 22:24

本文选题：甲基苯丙胺　切入点：大鼠依赖模型　出处：《昆明医科大学》2013年硕士论文

【摘要】：目的：建立甲基苯丙胺(MA)依赖大鼠模型,观察MA依赖鼠伏隔核和侧脑室注射胶质细胞源性营养因子(GDNF)后伏隔核多巴胺D2受体(DRD2)和多巴胺转运体(DAT)的变化,探讨外源性GDNF干预对甲基苯丙胺依赖鼠伏隔核机能代谢变化的影响。方法：SD大鼠160只,随机分为对照组、依赖一周、四周及八周组,建立不同依赖时间段的大鼠模型,三个实验组又分别分为5个亚组：单纯MA依赖组、MA依赖+假手术组、MA依赖+蒸馏水组、MA依赖+GDNF低剂量干预组及MA依赖+GDNF高剂量干预组。应用免疫组化技术、原位杂交技术、蛋白质免疫印迹技术和实时定量PCR技术检测伏隔核DRD2、DAT蛋白质及其mRNA的变化。结果：1.与对照组比较,三个单纯甲基苯丙胺依赖组的条件性位置偏爱实验均有显著性差异,甲基苯丙胺依赖大鼠模型成功建立。2.与对照组比较,三个实验组内不同亚组,伏隔核DRD2、DAT表达均有变化。DRD2及其mRNA在单纯MA依赖组、MA依赖+假手术组、MA依赖+蒸馏水组中表达均升高,而在MA依赖+GDNF低剂量干预组和MA依赖+GDNF高剂量干预组中表达降低；DAT及其mRNA在单纯MA依赖组MA依赖+假手术组、MA依赖+蒸馏水组中表达均降低,而MA依赖+GDNF低剂量干预组和MA依赖+GDNF高剂量干预组中表达明显回升。结论：1.采用多次腹腔注射甲基苯丙胺,结合条件性位置偏爱实验的方法成功建立甲基苯丙胺依赖大鼠模型。2.GDNF可改变甲基苯丙胺依赖大鼠伏隔核DRD2、DAT及其mRNA的表达,提示GDNF对甲基苯丙胺所致毒性作用有治疗保护作用。GDNF在0.5μg/μl和1.0μg/μl时均能有效改变甲基苯丙胺所致的DRD2及DAT及其mRNA的表达变化,其中在0.5μg/μl时的治疗干预效果较好。
[Abstract]:Aim: to establish a rat model of MA-dependent methamphetamine, and to observe the changes of dopamine D2 receptor (DRD2) and dopamine transporter (DATs) in nucleus accumbens and intracerebroventricular injection of glial cell-derived nutrition factor (GDNF) in MA-dependent rats.To investigate the effect of exogenous GDNF intervention on the metabolism of nucleus accumbens in methamphetamine-dependent rats.Methods 160 Sprague-Dawley rats were randomly divided into three groups: one week, four weeks and eight weeks dependent groups.The three experimental groups were divided into five subgroups: the MA dependent group, the MA dependent sham operation group, the MA dependent distilled water group, the MA dependent GDNF low dose intervention group, and the MA dependent GDNF high dose intervention group.The changes of DRD2DAT protein and its mRNA in nucleus accumbens were detected by immunohistochemistry, in situ hybridization, Western blot and real-time quantitative PCR.The result is 1: 1.Compared with the control group, there were significant differences among the three simple methamphetamine dependent groups in conditioned place preference (CPP) experiments, and the methamphetamine dependent rat model was established successfully.Compared with the control group, the expression of DRD2DAT in the nucleus accumbens and its mRNA were all increased in the MA dependent sham-operated group and the distilled water group.In MA dependent GDNF low dose intervention group and MA dependent GDNF high dose intervention group, the expression of mRNA was decreased in MA dependent sham operation group and distilled water group.However, the expression of MA-dependent low-dose GDNF and MA-dependent high-dose GDNF increased significantly.Conclusion 1.The rat model of methamphetamine dependent rats was successfully established by intraperitoneal injection of methamphetamine. 2. GDNF could change the expression of DRD2DAT and its mRNA in the nucleus accumbens of methamphetamine dependent rats.The results suggest that GDNF can effectively change the expression of DRD2, DAT and mRNA at 0.5 渭 g / 渭 l and 1.0 渭 g / 渭 l, especially at 0.5 渭 g / 渭 l.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：D919.1

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