丰富环境联合氟西汀干预对大鼠抑郁样行为、MBP及CNP的影响

发布时间：2018-01-04 01:30

  本文关键词：丰富环境联合氟西汀干预对大鼠抑郁样行为、MBP及CNP的影响　出处：《新乡医学院》2015年硕士论文　论文类型：学位论文

  更多相关文章： 丰富环境 氟西汀 髓鞘碱性蛋白 2

【摘要】：背景抑郁症已经成为全球重要的的公共卫生问题。研究表明抑郁症患者大脑前额叶、胼胝体、海马区等部位的脑白质的完整性明显缺损。脑白质有大量的神经纤维束组成,起着神经信号的传导作用。大量研究证实脑白质的损伤与抑郁症的发生密切相关。丰富环境可以增加脑白质有髓神经纤维总长度和总体积,还可以促进少突胶质细胞的增生,短期氟西汀治疗能够提高抑郁症患者脑白质的完整性。本研究建立慢性不可预见性应激(chronic unpredictable stress, CUS)刺激制备抑郁模型大鼠,使用丰富环境及氟西汀为干预方式,以前额叶、胼胝体、海马区为目标脑区,探讨丰富环境联合氟西汀干预对慢性应激所致抑郁大鼠行为学以及脑白质的影响。目的1.探讨丰富环境联合氟西汀干预对慢性应激所致抑郁大鼠行为学的影响。2.探讨丰富环境联合氟西汀干预对抑郁大鼠脑白质中髓鞘指标(髓鞘碱性蛋白)、以及少突胶质细胞标记物(2'-3’环核苷酸-3’-磷酸二酯酶)的影响。方法1.实验动物分组以及抑郁模型的建立SPF级雄性SD大鼠65只,采用旷场试验对所有大鼠评估,不合格的剔除。按照随机抽样的方法分配10只大鼠为正常对照(A)组,标准环境饲养。使用CUS法对剩余大鼠进行抑郁模型的建立。造模后随机抽样40只抑郁大鼠,分配为抑郁模型(B)组、氟西汀(C)组、丰富环境(D)组、丰富环境+氟西汀(E)组。2.氟西汀及丰富环境干预对于C组大鼠,给予抗抑郁药氟西汀10 mg/kg.d灌胃；对于D组大鼠,给予丰富环境12 h/d干预；对于E组大鼠,给予抗抑郁药氟西汀10 mg/kg.d灌胃,同时给予给予丰富环境12 h/d干预。3.行为学评估采用体质量的变化、蔗糖水消耗量实验、旷场实验三种方式分别在CUS刺激前、CUS刺激第21d,分别在氟西汀干预及丰富环境干预的第1周末、第2周末、第3周末评估大鼠的行为学变化。4. MBP、CNP蛋白含量及mRNA表达测定的实验方法使用免疫组化方法检测大鼠海马各区、前额叶区、胼胝体中MBP、CNP蛋白的含量；使用RT-PCR技术检测大鼠海马区、前额叶区MBP mRNA、CNP mRNA的表达量。5.统计方法实验数据用SPSS20.0软件包进行统计分析。多组计量资料采用单因素方差分析(one-way ANOVA);两两比较时,方差齐时用LSD法检验,方差不齐时用非参数独立样本Kruskal-Wallis检验分析,两组间均数比较采用独立样本t检验,配对t检验。P0.05为差异有显著性。结果1.行为学评估结果CUS刺激前,各组大鼠的体质量、旷场实验、蔗糖水消耗量的测量相比差异无统计学意义(P0.05)。经过21 d的CUS应激后；A组、C组、D组、E组大鼠与B组大鼠相比,在体质量变化、旷场实验、蔗糖水消耗量的测量上差异均有显著性(P0.05)。氟西汀及丰富环境干预第1周末,C组、D组、E组大鼠与B组大鼠相比部分行为明显改善,其中E组大鼠部分行为恢复较快；干预第2周末,E组大鼠与B组大鼠相比行为差异有显著性(P0.05)；干预第3周末；各组大鼠与B组大鼠相比行为差异均有显著性(P0.05),C组、D组、E组大鼠与A组大鼠相比行为差异均无显著性(P0.05),且这三组之间相比差异亦无显著性(P0.05)。2.大鼠海马、前额叶、胼胝体区MBP、CNP蛋白的含量结果在5组大鼠中,胼胝体区MBP、CNP蛋白的含量差异无显著性(P0.05)。在海马CA1区、DG区以及前额叶区五组大鼠之间的MBP的含量差异有显著性(F=6.956,P=0.000;F=6.098, P=0.001;F=6.590,P=0.000),其中B组大鼠MBP的含量与其它各组相比明显偏低,差异均有显著性(P0.05),且C组、D组、E组分别与A组相比差异均无显著性(P0.05)。在海马CA1区、DG区以及前额叶区CNP的含量差异有显著性(F=7.277,P=0.000;F=4.937, P=0.002; F=7.431, P=0.000)。其中B组大鼠MBP的含量与其它各组相比明显偏低,差异均有显著性(P0.05),C组、D组、E组分别与A组相比差异均无显著性(P0.05),且这三组之间相比差异亦无显著性(P0.05)。3.大鼠海马区、前额叶区MBP mRNA、CNP mRNA的表达量在5组大鼠中,海马区、前额叶区MBPmRNA的相对表达量差异有显著性(F=2.872,P=0.044;F=3.156, P=0.031),其中B组大鼠与其它各组大鼠相比MBPmRNA的相对表达量均显著降低(P0.05),且C组、D组、E组分别与A组相比差异均无显著性(P0.05)。海马区、前额叶区CNPmRNA的相对表达量均存在着明显的差异(F=4.283, P=0.009; F=2.829, P=0.046),其中B组大鼠与其它各组相比CNPmRNA的相对表达量显著降低,差异均有显著性(P0.05),C组、D组、E组大鼠分别与A组相比差异均无显著性(P0.05),且这三组之间相比差异亦无显著性(P0.05)。结论1.丰富环境和(或)氟西汀干预均可改善大鼠的抑郁样行为,丰富环境联合氟西汀在干预的早期表现出加快抑郁行为的改善。2.CUS所致抑郁模型大鼠的海马CA1、DG区及前额叶区MBP、CNP的蛋白含量及其转录水平均明显降低,可能与抑郁症的发病机制相关。3.丰富环境和(或)氟西汀均可提高海马CA1、DG区及前额叶区MBP、CNP的蛋白含量及其转录水平。
[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 琛屼负鎭㈠�嶈緝蹇�锛涘共棰勭��2鍛ㄦ湯,E缁勫ぇ榧犱笌B缁勫ぇ榧犵浉姣旇�屼负宸�寮傛湁鏄捐憲鎬，

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