本文關(guān)鍵詞：豐富環(huán)境聯(lián)合氟西汀干預對大鼠抑郁樣行為、MBP及CNP的影響　出處：《新鄉(xiāng)醫(yī)學院》2015年碩士論文　論文類型：學位論文

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【摘要】：背景抑郁癥已經(jīng)成為全球重要的的公共衛(wèi)生問題。研究表明抑郁癥患者大腦前額葉、胼胝體、海馬區(qū)等部位的腦白質(zhì)的完整性明顯缺損。腦白質(zhì)有大量的神經(jīng)纖維束組成,起著神經(jīng)信號的傳導作用。大量研究證實腦白質(zhì)的損傷與抑郁癥的發(fā)生密切相關(guān)。豐富環(huán)境可以增加腦白質(zhì)有髓神經(jīng)纖維總長度和總體積,還可以促進少突膠質(zhì)細胞的增生,短期氟西汀治療能夠提高抑郁癥患者腦白質(zhì)的完整性。本研究建立慢性不可預見性應(yīng)激(chronic unpredictable stress, CUS)刺激制備抑郁模型大鼠,使用豐富環(huán)境及氟西汀為干預方式,以前額葉、胼胝體、海馬區(qū)為目標腦區(qū),探討豐富環(huán)境聯(lián)合氟西汀干預對慢性應(yīng)激所致抑郁大鼠行為學以及腦白質(zhì)的影響。目的1.探討豐富環(huán)境聯(lián)合氟西汀干預對慢性應(yīng)激所致抑郁大鼠行為學的影響。2.探討豐富環(huán)境聯(lián)合氟西汀干預對抑郁大鼠腦白質(zhì)中髓鞘指標(髓鞘堿性蛋白)、以及少突膠質(zhì)細胞標記物(2'-3’環(huán)核苷酸-3’-磷酸二酯酶)的影響。方法1.實驗動物分組以及抑郁模型的建立SPF級雄性SD大鼠65只,采用曠場試驗對所有大鼠評估,不合格的剔除。按照隨機抽樣的方法分配10只大鼠為正常對照(A)組,標準環(huán)境飼養(yǎng)。使用CUS法對剩余大鼠進行抑郁模型的建立。造模后隨機抽樣40只抑郁大鼠,分配為抑郁模型(B)組、氟西汀(C)組、豐富環(huán)境(D)組、豐富環(huán)境+氟西汀(E)組。2.氟西汀及豐富環(huán)境干預對于C組大鼠,給予抗抑郁藥氟西汀10 mg/kg.d灌胃；對于D組大鼠,給予豐富環(huán)境12 h/d干預；對于E組大鼠,給予抗抑郁藥氟西汀10 mg/kg.d灌胃,同時給予給予豐富環(huán)境12 h/d干預。3.行為學評估采用體質(zhì)量的變化、蔗糖水消耗量實驗、曠場實驗三種方式分別在CUS刺激前、CUS刺激第21d,分別在氟西汀干預及豐富環(huán)境干預的第1周末、第2周末、第3周末評估大鼠的行為學變化。4. MBP、CNP蛋白含量及mRNA表達測定的實驗方法使用免疫組化方法檢測大鼠海馬各區(qū)、前額葉區(qū)、胼胝體中MBP、CNP蛋白的含量；使用RT-PCR技術(shù)檢測大鼠海馬區(qū)、前額葉區(qū)MBP mRNA、CNP mRNA的表達量。5.統(tǒng)計方法實驗數(shù)據(jù)用SPSS20.0軟件包進行統(tǒng)計分析。多組計量資料采用單因素方差分析(one-way ANOVA);兩兩比較時,方差齊時用LSD法檢驗,方差不齊時用非參數(shù)獨立樣本Kruskal-Wallis檢驗分析,兩組間均數(shù)比較采用獨立樣本t檢驗,配對t檢驗。P0.05為差異有顯著性。結(jié)果1.行為學評估結(jié)果CUS刺激前,各組大鼠的體質(zhì)量、曠場實驗、蔗糖水消耗量的測量相比差異無統(tǒng)計學意義(P0.05)。經(jīng)過21 d的CUS應(yīng)激后；A組、C組、D組、E組大鼠與B組大鼠相比,在體質(zhì)量變化、曠場實驗、蔗糖水消耗量的測量上差異均有顯著性(P0.05)。氟西汀及豐富環(huán)境干預第1周末,C組、D組、E組大鼠與B組大鼠相比部分行為明顯改善,其中E組大鼠部分行為恢復較快；干預第2周末,E組大鼠與B組大鼠相比行為差異有顯著性(P0.05)；干預第3周末；各組大鼠與B組大鼠相比行為差異均有顯著性(P0.05),C組、D組、E組大鼠與A組大鼠相比行為差異均無顯著性(P0.05),且這三組之間相比差異亦無顯著性(P0.05)。2.大鼠海馬、前額葉、胼胝體區(qū)MBP、CNP蛋白的含量結(jié)果在5組大鼠中,胼胝體區(qū)MBP、CNP蛋白的含量差異無顯著性(P0.05)。在海馬CA1區(qū)、DG區(qū)以及前額葉區(qū)五組大鼠之間的MBP的含量差異有顯著性(F=6.956,P=0.000;F=6.098, P=0.001;F=6.590,P=0.000),其中B組大鼠MBP的含量與其它各組相比明顯偏低,差異均有顯著性(P0.05),且C組、D組、E組分別與A組相比差異均無顯著性(P0.05)。在海馬CA1區(qū)、DG區(qū)以及前額葉區(qū)CNP的含量差異有顯著性(F=7.277,P=0.000;F=4.937, P=0.002; F=7.431, P=0.000)。其中B組大鼠MBP的含量與其它各組相比明顯偏低,差異均有顯著性(P0.05),C組、D組、E組分別與A組相比差異均無顯著性(P0.05),且這三組之間相比差異亦無顯著性(P0.05)。3.大鼠海馬區(qū)、前額葉區(qū)MBP mRNA、CNP mRNA的表達量在5組大鼠中,海馬區(qū)、前額葉區(qū)MBPmRNA的相對表達量差異有顯著性(F=2.872,P=0.044;F=3.156, P=0.031),其中B組大鼠與其它各組大鼠相比MBPmRNA的相對表達量均顯著降低(P0.05),且C組、D組、E組分別與A組相比差異均無顯著性(P0.05)。海馬區(qū)、前額葉區(qū)CNPmRNA的相對表達量均存在著明顯的差異(F=4.283, P=0.009; F=2.829, P=0.046),其中B組大鼠與其它各組相比CNPmRNA的相對表達量顯著降低,差異均有顯著性(P0.05),C組、D組、E組大鼠分別與A組相比差異均無顯著性(P0.05),且這三組之間相比差異亦無顯著性(P0.05)。結(jié)論1.豐富環(huán)境和(或)氟西汀干預均可改善大鼠的抑郁樣行為,豐富環(huán)境聯(lián)合氟西汀在干預的早期表現(xiàn)出加快抑郁行為的改善。2.CUS所致抑郁模型大鼠的海馬CA1、DG區(qū)及前額葉區(qū)MBP、CNP的蛋白含量及其轉(zhuǎn)錄水平均明顯降低,可能與抑郁癥的發(fā)病機制相關(guān)。3.豐富環(huán)境和(或)氟西汀均可提高海馬CA1、DG區(qū)及前額葉區(qū)MBP、CNP的蛋白含量及其轉(zhuǎn)錄水平。
[Abstract]:Background depression has become an important global public health problem. The research showed that depression in patients with prefrontal, corpus callosum, brain white matter integrity obvious defect in hippocampus area of brain white matter. There are a large number of nerve fiber bundles, plays a role in neural signal conduction. Many studies have confirmed that closely related to injury and depression the cerebral white matter. The enriched environment can increase the cerebral white matter and the total length of the myelinated fibers and the total volume, but also can promote oligodendrocyte proliferation, short-term fluoxetine therapy can improve the depression of patients with cerebral white matter integrity. This study established the chronic unpredictable stress (chronic unpredictable, stress, CUS) stimulate the preparation of rat model of depression, using the rich environment and fluoxetine as intervention before, frontal lobe, corpus callosum, hippocampus to target brain regions, the effects of enriched environment combined with fluoxetine intervention on As well as the effects of brain white matter behavior induced by chronic stress depression rats. Objective: 1. to investigate the effects of.2. combined with fluoxetine enriched environment intervention on chronic stress induced depression rat behavior of fluoxetine hydrochloride combined with enriched environment intervention on depression index of rat brain myelin in the white matter (myelin basic protein), and oligodendrocyte marker material (2'-3 '-3' - cyclic nucleotide phosphodiesterase). The effects of 1. methods of experimental animal model of depression group and SPF grade male SD 65 rats by open field test and evaluation of all rats, eliminate unqualified. According to the method of random sampling distribution of 10 rats as normal control (A) group, standard environment. The establishment of the remaining rats were in depression model using the CUS method. After modeling the random sampling 40 depression rats, distribution model of depression (B) group, fluoxetine (C) group, enriched environment (D) group, fluoxetine + enriched environment Ting (E).2. group and fluoxetine EE on C rats given antidepressant fluoxetine 10 mg/kg.d orally; the rats in group D, 12 h/d to give a rich environment for intervention; E rats given antidepressant fluoxetine 10 mg/kg.d orally, and given the rich environment of 12 h/d the intervention of.3. evaluation of the behavior change with body weight, sucrose consumption test, open field test in three ways respectively before CUS stimulation, the stimulation of CUS 21d, respectively, in the intervention of fluoxetine and enriched environment intervention at the end of the first week, second week, third week assessment rats behavioral changes of.4. MBP and CNP protein the experimental method for the determination of content and expression of mRNA using immunohistochemical method to detect the rat hippocampus, prefrontal cortex, corpus callosum in MBP, the content of CNP protein; using RT-PCR technique to detect rat hippocampus, prefrontal region of MBP mRNA, CNP mRNA scale up.5. statistics Experimental data were analyzed using SPSS20.0 software. Multiple sets of measurement data using single factor analysis of variance (one-way ANOVA); 22, when the LSD method is used to test the homogeneity of variance, homogeneity of variance in nonparametric independent sample Kruskal-Wallis test, the two groups were compared using independent samples t test, paired t.P0.05 test for significant differences. Results 1. behavior evaluation results before CUS stimulation, the rats body weight, open field test and sucrose consumption measurements had no significant difference (P0.05). After 21 d CUS stress; A group, C group, D group, E group rats compared with B rats, the changes of body mass, open field test, measurement of sucrose water consumption had significant difference (P0.05). Fluoxetine and enriched environment intervention for first weeks, C group, D group, E group rats compared with the behavior of rats in B group were significantly improved, which group E rats 琛屼負鎭㈠杈冨揩錛涘共棰勭2鍛ㄦ湯,E緇勫ぇ榧犱笌B緇勫ぇ榧犵浉姣旇涓哄樊寮傛湁鏄捐憲鎬，

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