急慢性应激大鼠多巴胺相关基因mRNA表达及miRNA表达谱的研究

发布时间：2018-09-03 15:46

【摘要】：目的探讨急慢性应激对抑郁动物模型伏隔核多巴胺通路相关基因表达的影响;探讨慢性不可预计温和应激模型miRNA表达谱差异。方法1.采用电击诱导急性应激模型,将16只SD大鼠随机分为正常对照组和急性应激组,每组8只,对照组常规饲养,不予任何处理,急性应激组每天予以40分钟足部电击,连续3天;采用慢性不可预计温和应激模型,将16只SD大鼠随机分为正常对照组和慢性应激组,每组8只,对照组常规饲养,不予任何处理,慢性应激组每天随机予1-2种温和刺激,共6周,每周称重,每3周测糖水偏爱性;造模结束后均采用旷场试验评价大鼠行为,安乐处死,取伏隔核低温保存。2.采用荧光实时定量PCR技术测定大鼠伏隔核多巴胺通路相关基因mRNA的表达。3.选取慢性不可预计温和应激试验中正常对照组和慢性应激组伏隔核样本各2例,采用miRNA测序技术比较两组大鼠miRNA表达差异。结果1.电击诱导的急性应激引起大鼠旷场试验行为减少。2.电击诱导的急性应激大鼠伏隔核多巴胺通路相关基因TH、DAT、DRD1、DRD2、DRD3、par-4的mRNA表达与对照组比较差异无统计学意义。3.慢性不可预计温和应激引起大鼠体重增长(240.91±27.59g vs 156.30±15.78g)和糖水偏爱度(.73±.10vs.60±.14)显著下降,旷场试验行为增多。4.慢性不可预计温和应激大鼠伏隔核多巴胺通路相关基因TH、DAT、DRD1、DRD2、DRD3、par-4的mRNA表达与对照组比较差异无统计学意义。5.慢性不可预计温和应激大鼠与正常对照组比较25个miRNA显著下调;2个miRNA显著上调。结论1.电击诱导的急性应激和慢性不可预计温和应激可能不影响伏隔核多巴胺通路相关基因TH、DAT、DRD1、DRD2、DRD3、par-4的mRNA表达。2.慢性不可预计温和应激可能使大鼠部分miRNA表达发生改变。
[Abstract]:Objective to investigate the effect of acute and chronic stress on the expression of dopamine pathway related genes in nucleus accumbens of depressive animal model and to explore the difference of miRNA expression profile in chronic unpredictable mild stress model. Method 1. Sixteen SD rats were randomly divided into normal control group (n = 8) and acute stress group (n = 8). Sixteen SD rats were randomly divided into normal control group (n = 8) and chronic stress group (n = 8). The rats were weighed every week for 6 weeks, and the preference for sugar water was measured every 3 weeks. The behavior of the rats was evaluated by open field test after the model was made, the rats were killed in euthanasia, and the nucleus accumbens was preserved by hypothermia. The expression of dopamine pathway related gene (mRNA) in rat nucleus accumbens was determined by fluorescence real time quantitative PCR. In the chronic unpredictable mild stress test, two samples of nucleus accumbens were collected from normal control group and two cases of chronic stress group, and the difference of miRNA expression between the two groups was compared by miRNA sequencing technique. Result 1. Acute stress induced by electric shock reduced open field test behavior in rats. 2. 2. The mRNA expression of dopamine pathway related gene TH,DAT,DRD1,DRD2,DRD3,par-4 in nucleus accumbens of acute stress rats induced by electric shock was not significantly different from that of control group. Chronic unpredictable mild stress significantly decreased weight gain (240.91 卤27.59g vs 156.30 卤15.78g) and sugar water preference (.73 卤.10vs.60 卤.14), and increased open-field test behavior. There was no significant difference in mRNA expression of dopamine pathway related gene TH,DAT,DRD1,DRD2,DRD3,par-4 between chronic unpredictable mild stress rats and control group. Compared with the control group, 25 miRNA and 2 miRNA were significantly down-regulated and up-regulated in chronic unpredictable mild stress rats. Conclusion 1. Electric shock induced acute stress and chronic unpredictable mild stress may not affect the expression of mRNA. 2. 2 of dopamine pathway related gene TH,DAT,DRD1,DRD2,DRD3,par-4 in nucleus accumbens. Chronic unpredictable mild stress may change the expression of partial miRNA in rats.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.4

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