天麻素干预甲基苯丙胺依赖大鼠后相关脑区谷氨酸受体、转运体的表达研究
发布时间：2018-08-02 12:07
【摘要】：目的:建立甲基苯丙胺(MA)依赖大鼠模型,使用天麻素进行干预,检测谷氨酸受体(Grin2B、Glur-1)、谷氨酸转运体(GLT-1、EAAT-1)及凋亡因子Caspase-3在MA依赖大鼠六个相关脑区(额叶皮质、纹状体、伏隔核、海马、黑质、中脑腹侧被盖区)的表达,探讨天麻素对甲基苯丙胺所致中枢神经系统毒性损害是否具有保护作用。方法:SD大鼠80只,随机将80只大鼠分为6周组及8周组。将每组再分为:空白对照组,天麻素前干预组,甲基苯丙胺依赖组,天麻素(10mg)干预治疗组及天麻素(20mg)干预治疗组。通过腹腔注射甲基苯丙胺,采用条件性位置偏爱实验(CPP)和刻板行为评分观察行为学变化,建立甲基苯丙胺依赖大鼠模型。确认大鼠产生甲基苯丙胺依赖后,建成不同时间段的依赖模型,注射不同剂量的天麻素(10mg或20mg)干预。应用免疫荧光、蛋白免疫印迹技术和实时定量PCR技术检测鼠脑不同脑区谷氨酸受体(Grin2B、Glur-1)、谷氨酸转运体(GLT-1、EAAT-1)及 Caspase-3 的表达变化。结果:1、与对照组相比,MA依赖组和天麻素干预各组的条件性位置偏爱实验结果均有显著性差异,表明成功建立了甲基苯丙胺依赖大鼠模型。2、MA依赖组Caspase-3表达增多;4周MA依赖组NR2B表达增高;大部分脑区Glur-1表达量增高;2周MA依赖组GLT-1、EAAT-1表达量降低,4周MA依赖组GLT-1、EAAT-1表达量增高。3、与天麻素干预前比较,天麻素干预后大鼠在伴药盒停留时间明显缩短。刻板行为评分,3分及4分次数明显减少。4、天麻素干预后对MA依赖组Caspase-3、谷氨酸受体(Grin2B、Glur-1)及谷氨酸转运体(GLT-1、EAAT-1)的增高有治疗作用,且天麻素20mg干预组效果优于10mg组。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和刻板行为评分的方法成功建立甲基苯丙胺依赖大鼠模型。2、与对照组比较,除谷氨酸转运体(GLT-1、EAAT-1)在2周MA依赖组降低,Caspase-3、谷氨酸受体(Grin2B、Glur-1)及谷氨酸转运体(GLT-1、EAAT-1)表达总体呈增高趋势,提示Caspase-3、谷氨酸受体(Grin2B、Glur-1)及谷氨酸转运体(GLT-1、EAAT-1)与MA神经毒性和依赖机制密切相关。3、天麻素干预后对MA依赖组大鼠行为学指标有改善,对Caspase-3、谷氨酸受体(Grin2B、Glur-1)及谷氨酸转运体(GLT-1、EAAT-1)的增高有一定改善作用,且天麻素20mg干预组效果优于10mg组。提示天麻素对甲基苯丙胺所致中枢神经系统毒性损害具有保护作用。
[Abstract]:Aim: to establish a methamphetamine (MA) dependent rat model, and to investigate the effects of Gastrodin on glutamate receptor, glutamate transporter (GLT-1EAAT-1) and apoptosis factor Caspase-3 in six brain regions (frontal cortex, striatum, nucleus accumbens) of MA-dependent rats. The expression of Gastrodin in hippocampus, substantia nigra and ventral tegmental area of the midbrain, to investigate the protective effect of Gastrodin on the central nervous system (CNS) toxicity induced by methamphetamine. Methods 80 Sprague-Dawley rats were randomly divided into 6 week group and 8 week group. Each group was divided into three groups: blank control group, Gastrodin preintervention group, methamphetamine dependent group, 10mg intervention group and 20mg intervention group. A model of methamphetamine dependent rats was established by intraperitoneal injection of methamphetamine. The behavioral changes were observed by using conditioned place preference test (CPP) and stereotypical behavior score. After methamphetamine dependence was produced in rats, different time dependent models were established and different doses of Gastrodin (10mg or 20mg) were injected. The expression of glutamate receptor (Grin2BmGlur-1), glutamate transporter (GLT-1EAAT-1) and Caspase-3 in different brain regions were detected by immunofluorescence, Western blot and real-time quantitative PCR. Results compared with the control group, there were significant differences in the experimental results of conditioned place preference between the MA dependent group and Gastrodin intervention group. The results showed that the expression of Caspase-3 was increased in the methamphetamine dependent group and the NR2B expression was increased in the MA dependent group after 4 weeks. The expression of GLT-1 + EAAT-1 decreased in the MA dependent group at 2 weeks. The expression of GLT-1 + EAAT-1 was significantly increased in the MA dependent group at 4 weeks. Compared with that before Gastrodin intervention, the residence time of Gastrodin group was significantly shorter than that before Gastrodin intervention. The scores of stereotypical behavior, scores of 3 and times of 4 scores were significantly decreased. After the intervention of Gastrodin, the increase of Caspase-3, glutamate receptor (Grin2BtGlur-1) and glutamate transporter (GLT-1EAAT-1) in MA dependent group was significantly increased, and the effect of Gastrodin 20mg intervention group was better than that of 10mg group. ConclusionThe methamphetamine dependent rat model was successfully established by multiple intraperitoneal injection of methamphetamine, combined with conditional place preference test and stereotypical behavior score, compared with the control group. The expression of glutamate transporter (GLT-1) and glutamate transporter (GLT-1EAAT-1) decreased significantly in MA-dependent group at 2 weeks, and the expression of glutamate receptor (Grin2BtGlur-1) and glutamate transporter (GLT-1EAAT-1) increased significantly in MA-dependent group. These results suggest that Caspase-3, glutamate receptor (Grin2BnGlur-1) and glutamate transporter (GLT-1) are closely related to the neurotoxicity and dependence mechanism of MA. The increase of Caspase-3, glutamate receptor (Grin2BtGlur-1) and glutamate transporter (GLT-1) were improved to some extent, and the effect of Gastrodin 20mg intervention group was better than that of 10mg group. The results suggest that Gastrodin has protective effect on central nervous system toxicity induced by methamphetamine.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.64
[Abstract]:Aim: to establish a methamphetamine (MA) dependent rat model, and to investigate the effects of Gastrodin on glutamate receptor, glutamate transporter (GLT-1EAAT-1) and apoptosis factor Caspase-3 in six brain regions (frontal cortex, striatum, nucleus accumbens) of MA-dependent rats. The expression of Gastrodin in hippocampus, substantia nigra and ventral tegmental area of the midbrain, to investigate the protective effect of Gastrodin on the central nervous system (CNS) toxicity induced by methamphetamine. Methods 80 Sprague-Dawley rats were randomly divided into 6 week group and 8 week group. Each group was divided into three groups: blank control group, Gastrodin preintervention group, methamphetamine dependent group, 10mg intervention group and 20mg intervention group. A model of methamphetamine dependent rats was established by intraperitoneal injection of methamphetamine. The behavioral changes were observed by using conditioned place preference test (CPP) and stereotypical behavior score. After methamphetamine dependence was produced in rats, different time dependent models were established and different doses of Gastrodin (10mg or 20mg) were injected. The expression of glutamate receptor (Grin2BmGlur-1), glutamate transporter (GLT-1EAAT-1) and Caspase-3 in different brain regions were detected by immunofluorescence, Western blot and real-time quantitative PCR. Results compared with the control group, there were significant differences in the experimental results of conditioned place preference between the MA dependent group and Gastrodin intervention group. The results showed that the expression of Caspase-3 was increased in the methamphetamine dependent group and the NR2B expression was increased in the MA dependent group after 4 weeks. The expression of GLT-1 + EAAT-1 decreased in the MA dependent group at 2 weeks. The expression of GLT-1 + EAAT-1 was significantly increased in the MA dependent group at 4 weeks. Compared with that before Gastrodin intervention, the residence time of Gastrodin group was significantly shorter than that before Gastrodin intervention. The scores of stereotypical behavior, scores of 3 and times of 4 scores were significantly decreased. After the intervention of Gastrodin, the increase of Caspase-3, glutamate receptor (Grin2BtGlur-1) and glutamate transporter (GLT-1EAAT-1) in MA dependent group was significantly increased, and the effect of Gastrodin 20mg intervention group was better than that of 10mg group. ConclusionThe methamphetamine dependent rat model was successfully established by multiple intraperitoneal injection of methamphetamine, combined with conditional place preference test and stereotypical behavior score, compared with the control group. The expression of glutamate transporter (GLT-1) and glutamate transporter (GLT-1EAAT-1) decreased significantly in MA-dependent group at 2 weeks, and the expression of glutamate receptor (Grin2BtGlur-1) and glutamate transporter (GLT-1EAAT-1) increased significantly in MA-dependent group. These results suggest that Caspase-3, glutamate receptor (Grin2BnGlur-1) and glutamate transporter (GLT-1) are closely related to the neurotoxicity and dependence mechanism of MA. The increase of Caspase-3, glutamate receptor (Grin2BtGlur-1) and glutamate transporter (GLT-1) were improved to some extent, and the effect of Gastrodin 20mg intervention group was better than that of 10mg group. The results suggest that Gastrodin has protective effect on central nervous system toxicity induced by methamphetamine.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.64
【参考文献】
相关期刊论文 前10条
1 李俊葵;刘伟;彭菊;江明金;刘怡;郭友立;莫志贤;;钩藤碱对甲基苯丙胺依赖小鼠位置偏爱和海马谷氨酸受体1表达的影响[J];中国实验方剂学杂志;2014年18期
2 赵瑞珍;唐启盛;田青;李小黎;;抑郁大鼠脑内NMDA受体NR1、NR2B表达及中药干预作用[J];北京中医药大学学报;2012年06期
3 柳星;汪萌芽;;天麻素对海马内Aβ_(1-40)注射阿尔茨海默病模型大鼠学习记忆的改善作用[J];中国临床药理学与治疗学;2012年04期
4 赵敏;郝伟;;苯丙胺类兴奋剂所致精神障碍的临床诊治问题[J];上海精神医学;2011年06期
5 段晓飞;邓冲;曾晓锋;赵永和;王尚文;李桢;;甲基苯丙胺中毒大鼠相关脑区Caspase-3和Bcl-2表达的研究[J];现代生物医学进展;2011年03期
6 李海龙;粟艳;厚荣荣;陈宏;陈建宗;;天麻素对百草枯和代森锰诱导的小鼠脑黑质多巴胺能神经元损伤的保护作用[J];成都中医药大学学报;2010年01期
7 曾晓锋;路钢;李桢;;甲基苯丙胺致多巴胺能神经元毒性机制的研究进展[J];中国法医学杂志;2010年01期
8 赵燕玲;张博爱;贾延R，
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