睡眠剥夺对大鼠不同脑区5羟色胺1A受体、多巴胺受体基因表达的影响

发布时间：2018-03-26 00:11

本文选题：大鼠　切入点：睡眠剥夺　出处：《新乡医学院》2015年硕士论文

【摘要】：背景现有证据表明,在睡眠剥夺期间与清醒期间有着不同的基因表达。睡眠剥夺对5-HT、DA等神经递质的改变已有相关的研究,但它们之间的平衡转换和协同作用的调节机制目前尚不清楚。目的通过检测大鼠在不同时间睡眠剥夺三个脑区(海马、下丘脑、纹状体)5-HT受体1A、DA受体D1基因和DA受体D2基因表达改变情况,分析睡眠剥夺对这些基因表达的影响,探讨不同神经递质通路参与睡眠调节的差异。方法1选择10周龄雄性SD大鼠24只。分别进行24小时、48小时、72小时REM睡眠剥夺。2采用旷场实验检测动物潜伏期、水平分数、直立次数。验证睡眠剥夺体力和精神疲劳模型。3每组取大鼠海马、下丘脑、纹状体三个脑区脑组织,利用RT-PCR技术分别检测5羟色胺1A受体和多巴胺D1R、D2R基因的表达情况。4应用SPSS11.5统计软件对实验数据进行相应的统计与处理。计量资料进行统计分析选用均数±标准差(x±s)。两组间均数进行对比与分析采用独立样本t验；多组间均数进行比较采用单因素方差分析,进行两两比较采用LSD检验,进行相关性分析采用Pearson相关,P0.05表示统计结果的差异具有显著性。结果1 SD大鼠经过睡眠剥夺后进行旷场实验,水平得分及垂直得分均有很大的波动,大鼠对环境的认知能力明显下降,体力、精神疲劳。说明睡眠剥夺对机体的精神活动产生抑制。2睡眠剥夺影响了大鼠海马(F=56.203,P=0.000)和纹状体(F=77.288,P=0.000)区域5-HT1A的基因表达,差异具有统计学意义,在海马和纹状体区域各睡眠剥夺组大鼠的表达量均高于对照组,差异具有统计学意义(P0.05),在海马剥夺72小时组高于剥夺24小时(P=0.005)和48小时(P=0.010)组,纹状体区域则各组动物的差异均具有统计学意义(P0.05)。各组大鼠下丘脑区域5-HT1A的基因表达差异不具有统计学意义(P0.05)。3睡眠剥夺影响了大鼠海马(F=74.708,P=0.000)、纹状体(F=80.687,P=0.000)区域D2基因的表达,差异具有统计学意义,各睡眠剥夺组大鼠在海马和纹状体区域的表达量均低于对照组,差异具有统计学意义(P0.05),剥夺时间长的动物基因表达量也更低,海马与纹状体脑区基因表达量各组动物的差异均具有统计学意义(P均0.05)。各组大鼠下丘脑区域D2R的基因表达表达差异不具有统计学意义(P0.05)。4相关分析显示对照组各脑区5-HT1A和D2R基因的表达不存在相关性,睡眠剥夺24小时在海马(r=-0.868,P=0.025)、纹状体(r=-0.883,P=0.02)、下丘脑(r=-0.841,P=0.036)两个基因的表达均呈负相关,睡眠剥夺48小时和72小时在海马(r=-0.881,P=0.02；r=-0.878,P=0.021)、纹状体(r=-0.735,P=0.006；r=-0.855,P=0.03)两个基因的表达也呈负相关。5睡眠剥夺对大鼠海马、纹状体和下丘脑区域D1R基因表达表达差异不具有统计学意义(P0.05)。小结睡眠剥夺大鼠在海马和纹状体5-HT1A与D2R基因表达均发生显著变化,5-HT1A基因表达增高、D2R基因表达下降,并呈负相关,表明5-羟色胺系统和多巴胺系统在受体水平对睡眠节律的调节具有协同性。
[Abstract]:The background of the current evidence shows that there is in sleep deprivation during the waking period and different gene expression. Sleep deprivation on 5-HT, on DA related neurotransmitter changes already, but the regulatory mechanism of balance conversion and synergy among them is unclear. Objective to detect rats in sleep deprivation three brain regions (hippocampus, hypothalamus, striatum) 5-HT receptor 1A, DA receptor expression of D1 gene and DA receptor D2 gene changes, analysis of effects of sleep deprivation on the expression of these genes, the differences between different neurotransmitter pathways involved in sleep regulation. Methods: 1 10 week old male SD 24 rats were 24. Hours, 48 hours, 72 hours of sleep deprivation REM.2 using the open field test animal level scores, rearing latency, sleep deprivation. Verify the physical and mental fatigue model.3 from each rat hippocampus, hypothalamus, striatum The brain of three brain regions, the use of RT-PCR technology to detect 5 serotonin 1A receptor and dopamine D1R, D2R gene expression of.4 using SPSS11.5 statistical software for statistical processing and corresponding to the experimental data. The measurement data were used for statistical analysis the mean and standard deviation (x + s) between the two groups were analyzed. Comparison and analysis by using independent sample t test; multiple groups were compared using one-way ANOVA, 22 were compared with LSD test and correlation analysis using Pearson correlation, P0.05 said the difference was significant. The statistical results of 1 SD rats after sleep deprivation after the open field test scores and level fluctuation the vertical score significantly, cognitive ability of rats decreased significantly in the environment, physical and mental fatigue. It inhibited the effect of.2 sleep deprivation rat hippocampus on the mental activity of sleep deprivation (F=56. 203, P=0.000) and striatum (F=77.288, P=0.000) expression region of 5-HT1A gene, there was significant difference in the expression of hippocampus and striatum of the sleep deprivation group rats were higher than the control group, the difference was statistically significant (P0.05) in the hippocampus, 72 hours deprivation group was higher than that of deprivation for 24 hours (P=0.005) and 48 h (P=0.010) group, the difference of each animal striatum were statistically significant (P0.05). Each group the hypothalamus 5-HT1A rat gene expression difference was not statistically significant (P0.05).3 sleep deprivation affects the hippocampus (F=74.708, P=0.000), striatum (F=80.687, P=0.000) expression of D2 gene region, the difference was statistically significant, the sleep deprivation group rats were lower than the control group in the expression of hippocampus and striatum volume, the difference was statistically significant (P0.05), deprived of animal gene expression is lower in long time, The difference of gene expression in hippocampus and striatum area of each animal were statistically significant (P < 0.05). Each region of the hypothalamus of rat D2R gene expression difference was not statistically significant (P0.05).4 correlation analysis showed that 5-HT1A and D2R control group of brain gene expression is associated with sleep deprivation for 24 hours (r=-0.868, P=0.025) in the hippocampus, striatum, hypothalamus (r=-0.883, P=0.02) (r=-0.841, P=0.036) two gene expression was negatively related to sleep deprivation for 48 hours and 72 hours in the hippocampus (r=-0.881, P=0.02; r= -0.878, P=0.021), striatum (r=-0.735, P=0.006; r=-0.855, P=0.03) two the gene was also negatively related to.5 sleep deprivation on rat hippocampus, striatum and hypothalamus D1R gene expression difference was not statistically significant (P0.05). Summary of sleep deprivation rats in the hippocampus and striatum of 5-HT1A and D2R gene All of them had significant changes. The expression of 5-HT1A gene increased, and the expression of D2R gene decreased. It was negatively correlated, indicating that 5- serotonin system and dopamine system play a synergistic role in the regulation of sleep rhythm at the receptor level.

【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R740

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