【摘要】：目的探討急慢性應(yīng)激對(duì)抑郁動(dòng)物模型伏隔核多巴胺通路相關(guān)基因表達(dá)的影響;探討慢性不可預(yù)計(jì)溫和應(yīng)激模型miRNA表達(dá)譜差異。方法1.采用電擊誘導(dǎo)急性應(yīng)激模型,將16只SD大鼠隨機(jī)分為正常對(duì)照組和急性應(yīng)激組,每組8只,對(duì)照組常規(guī)飼養(yǎng),不予任何處理,急性應(yīng)激組每天予以40分鐘足部電擊,連續(xù)3天;采用慢性不可預(yù)計(jì)溫和應(yīng)激模型,將16只SD大鼠隨機(jī)分為正常對(duì)照組和慢性應(yīng)激組,每組8只,對(duì)照組常規(guī)飼養(yǎng),不予任何處理,慢性應(yīng)激組每天隨機(jī)予1-2種溫和刺激,共6周,每周稱重,每3周測(cè)糖水偏愛性;造模結(jié)束后均采用曠場(chǎng)試驗(yàn)評(píng)價(jià)大鼠行為,安樂處死,取伏隔核低溫保存。2.采用熒光實(shí)時(shí)定量PCR技術(shù)測(cè)定大鼠伏隔核多巴胺通路相關(guān)基因mRNA的表達(dá)。3.選取慢性不可預(yù)計(jì)溫和應(yīng)激試驗(yàn)中正常對(duì)照組和慢性應(yīng)激組伏隔核樣本各2例,采用miRNA測(cè)序技術(shù)比較兩組大鼠miRNA表達(dá)差異。結(jié)果1.電擊誘導(dǎo)的急性應(yīng)激引起大鼠曠場(chǎng)試驗(yàn)行為減少。2.電擊誘導(dǎo)的急性應(yīng)激大鼠伏隔核多巴胺通路相關(guān)基因TH、DAT、DRD1、DRD2、DRD3、par-4的mRNA表達(dá)與對(duì)照組比較差異無統(tǒng)計(jì)學(xué)意義。3.慢性不可預(yù)計(jì)溫和應(yīng)激引起大鼠體重增長(240.91±27.59g vs 156.30±15.78g)和糖水偏愛度(.73±.10vs.60±.14)顯著下降,曠場(chǎng)試驗(yàn)行為增多。4.慢性不可預(yù)計(jì)溫和應(yīng)激大鼠伏隔核多巴胺通路相關(guān)基因TH、DAT、DRD1、DRD2、DRD3、par-4的mRNA表達(dá)與對(duì)照組比較差異無統(tǒng)計(jì)學(xué)意義。5.慢性不可預(yù)計(jì)溫和應(yīng)激大鼠與正常對(duì)照組比較25個(gè)miRNA顯著下調(diào);2個(gè)miRNA顯著上調(diào)。結(jié)論1.電擊誘導(dǎo)的急性應(yīng)激和慢性不可預(yù)計(jì)溫和應(yīng)激可能不影響伏隔核多巴胺通路相關(guān)基因TH、DAT、DRD1、DRD2、DRD3、par-4的mRNA表達(dá)。2.慢性不可預(yù)計(jì)溫和應(yīng)激可能使大鼠部分miRNA表達(dá)發(fā)生改變。
[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 鹵27.59g vs 156.30 鹵15.78g) and sugar water preference (.73 鹵.10vs.60 鹵.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.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R749.4

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