条件性位置厌恶大鼠伏隔核壳区多巴胺及DOPAC浓度变化

发布时间：2018-03-30 09:51

本文选题：吗啡依赖　切入点：条件性位置厌恶　出处：《新乡医学院》2013年硕士论文

【摘要】：目的通过测定吗啡依赖大鼠纳洛酮催瘾CPA建立、消退及重建不同阶段伏隔核壳区多巴胺(dopamine)及其代谢产物3,4二羟基苯乙酸(dihydroxyphenyl acetic acid, DOPAC)浓度的变化情况,探讨吗啡依赖负性强化的神经生化机制。方法 1、69只SPF级雄性(Sprage-Dawley)大鼠随机分为实验组(吗啡+纳洛酮,MN组)与对照组(吗啡+生理盐水,MS组;生理盐水+纳洛酮,SN组),每组23只,采用连续6.5天吗啡(10mg/kg,bid,IP)注射后进行一次纳洛酮催瘾(0.3mg/kg,IP),同时搭配条件性位置训练箱建立CPA大鼠模型,从day7下午至day13上午,对CPA大鼠进行每天2次的消退行为训练,第14天,对CPA行为消退了的大鼠进行药物“点燃”重建； 2、分别在套管针植入术后3-5天,CPA建立前、后及消退与重建后用微透析方法收集伏隔核壳区的细胞外液；采用高效液相色谱-电化学方法检测透析液中DA及DOPAC浓度。 3、所有结果如非特殊说明,均以均数±标准误(Mean±SEM)表示,多组均数间的多重比较,采用方差分析(ANOVA),组内行为学和不同时间点神经递质的两两比较采用Independent-Samples t-test。当P0.05被认为差异具有统计学意义。所有的统计分析均采用SPSS16.0软件完成。结果 1、连续6天半的吗啡注射与纳洛酮一次催瘾搭配可以成功建立CPA模型,实验鼠表现出明显的厌恶动机,对伴药侧表现出明显的回避；经过连续7天的消退训练后CPA行为可以成功消退；而消退后的CPA大鼠经药物“点燃”并纳洛酮催瘾后CPA行为可成功重建; 2、MN组大鼠伏隔核壳区多巴胺及DOPAC水平在CPA建立后明显升高(P0.01),而MS组与SN组较建立前差异无统计学意义(P0.05)；经过为期一周的消退训练之后,条件戒断大鼠DA/DOPAC水平明显下降至基础水平;CPA行为重建后,MN组大鼠伏隔核壳区的DA/DOPAC水平明显升高,与MS组、SN组相比差异具有显著统计学意义(P0.01)。结论 1、小剂量纳洛酮可以诱发吗啡依赖大鼠CPA的产生,经过为期一周的无药状态下消退行为训练,CPA行为可以成功消退,当再次暴露于药物相关环境时,消退了的CPA行为可以重建; 2、伏隔核壳区的多巴胺与其代谢产物DOPAC可能参与小剂量纳洛酮所诱发的吗啡依赖大鼠的条件性位置厌恶,中脑边缘系统的多巴胺机制可能在吗啡依赖的厌恶动机方面起到十分重要的调节作用。
[Abstract]:Purpose. The changes of dopamine dopamine (dopamine) and its metabolite, 3O4 dihydroxyphenyl acetic acid3 (DOPAC) in the nucleus putamen region of nucleus accumbens in morphine dependent rats induced by naloxone were determined. To explore the neurobiochemical mechanism of morphine dependent negative enhancement. Method. 1Sixty-nine SPF male Sprage-Dawley rats were randomly divided into two groups: the experimental group (morphine naloxone MN group) and the control group (morphine saline group MS), and the saline naloxone SN group with 23 rats in each group. The rats of CPA were induced by naloxone injection of 10 mg / kg IP for 6.5 days after injection of naloxone (0.3 mg / kg). The model of CPA rats was established by matching the conditioned position training box. From the afternoon of day7 to the morning of day13, the rats of CPA were trained twice a day in regression behavior, and the rats were trained twice a day on the 14th day. The drug "kindling" was performed on the rats whose CPA behavior disappeared. 2. Extracellular fluids in the nucleus and shell of accumbens were collected by microdialysis before and after the establishment of CPA at 3-5 days after cannula implantation, and the concentrations of DA and DOPAC in dialysis solution were detected by high performance liquid chromatography (HPLC) and electrochemical method. 3. All results, if not specified, are expressed as mean 卤standard error mean 卤SEM, multiple comparisons between multiple groups of mean, ANOVAN was used to compare behavior and neurotransmitters at different time points with Independent-Samples t-test.When P0.05 was considered to be statistically significant, all statistical analyses were performed by SPSS16.0 software. Results. 1. Six and a half days of morphine injection combined with naloxone can successfully establish the CPA model. The experimental mice showed obvious aversion motivation and obvious avoidance to the side of the drug. After 7 days of continuous regression training, the CPA behavior could be successfully dissipated, while the subsided CPA rats could be successfully reconstructed after drug "kindling" and naloxone-induced CPA behavior. (2) the levels of dopamine and DOPAC in the nucleus putamen of accumbent septum of rats in MN group increased significantly after the establishment of CPA, but there was no significant difference between group MS and group SN before establishment, and after one week of regression training, there was no significant difference between MS group and SN group. The level of DA/DOPAC was significantly decreased to the basic level in conditioned abstinence rats. The level of DA/DOPAC in the nucleus putamen of the accumbent septum in MN group was significantly higher than that in the SN group of MS, and there was a significant difference between MN group and MS group (P 0.01). Conclusion. 1. Low dose naloxone could induce the production of CPA in morphine dependent rats. After a week's training of regression behavior in drug-free state, the behavior of CPA could be successfully dissipated, and the subsided CPA behavior could be reconstructed when exposed to drug-related environment again. 2. Dopamine in the nucleus and shell of accumbent septum and its metabolite DOPAC may be involved in the conditioned positional aversion of morphine dependent rats induced by low dose naloxone. The dopamine mechanism in the midbrain limbic system may play an important role in the aversion motivation of morphine dependence.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.6

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