小鼠海马齿状回过表达18kDa转位蛋白对焦虑和抑郁样行为的调节

发布时间：2018-03-27 22:14

本文选题：海马齿状回　切入点：转位蛋白(TSPO)　出处：《河北北方学院》2017年硕士论文

【摘要】：18k Da转位蛋白(18k Da translocator protein,TSPO)在中枢神经系统中,TSPO主要分布在胶质细胞线粒体外膜上,能够介导胆固醇从线粒体外膜向线粒体内转运,参与神经类固醇合成的限速步骤,介导神经类固醇合成尤其是四氢孕酮的合成,而神经类固醇作为神经调节递质,影响γ-氨基丁酸,谷氨酸受体、等多个神经递质及其受体功能,在应激反应或焦虑,抑郁,创伤后应激障碍等应激相关疾病中起到重要的调节作用。近年来研究发现,TSPO可能成为情感性精神疾病尤其是焦虑症抑郁症治疗的潜在靶标。本研究旨在探究小鼠海马齿状回18k Da转位蛋白(TSPO)过表达对焦虑,抑郁样行为的影响,为TSPO成为药物潜在新靶标提供实验依据。本研究成年雄性ICR小鼠海马齿状回微注射携带TSPO基因的慢病毒,以致TSPO过表达,观察小鼠海马齿状回区注射慢病毒后注射位置荧光蛋白表达准确性以及TSPO蛋白表达水平的变化;第12,14和16天进行小鼠高架十字迷宫、爬梯实验、明暗箱实验检测焦虑样行为,第18,和21天进行小鼠悬尾和强迫游泳两个行为绝望模型检测抑郁样行为;同时采用Western Blot和ELISA方法分别检测海马注射孔周围3mm组织内TSPO蛋白水平以及四氢孕酮含量,初步探讨海马齿状回过表达TSPO与焦虑,抑郁障碍的关系。本研究结果如下:慢病毒定位注射后12天,过表达组小鼠海马组织内TSPO蛋白水平明显高于空白组(P=0.001);注射后12天,过表达组小鼠海马齿状回区绿色荧光蛋白明显;小鼠海马齿状回微注射慢病毒后12天,对小鼠自发活动无显著影响(P0.05)。与LV-NC组相比,LV-TSPO组显著增加小鼠进入中心区时间(P0.05);显著增加小鼠进入开臂次数和开臂时间的百分比(P0.05;P0.01);显著减少小鼠站立次数(P0.05);显著增加小鼠进入明箱的次数和时间(P0.05;P0.01),提示海马齿状回TSPO过表达小鼠表现出显著的抗焦虑样行为作用;与LV-NC组相比,LV-TSPO组明显减少小鼠在悬尾实验和强迫游泳实验中的不动时间(P0.001;P0.05),提示海马齿状回TSPO过表达小鼠表现出显著的抗抑郁样行为作用。进一步的生化检测表明,与阴性对照组相比,小鼠海马齿状回TSPO表达可导致海马注射孔周围3mm组织的TSPO蛋白表达水平显著增加(P0.05),并且其四氢孕酮含量显著增加(P0.05)。小鼠海马齿状回过表达TSPO具有显著的抗焦虑、抗抑郁样的行为学效应,且TSPO介导的四氢孕酮水平升高可能是其重要机制。
[Abstract]:In the central nervous system (CNS), the 18kDa translocation protein (18kDa translocator) mainly distributes on the mitochondrial outer membrane of glial cells, which can mediate the transport of cholesterol from the mitochondrial outer membrane to the mitochondria, and participate in the rate-limiting steps of neurosteroid synthesis. Mediates the synthesis of neurosteroids, especially tetrahydroprogesterone, and neurosteroids, as neuroregulatory transmitters, affect the functions of many neurotransmitters, such as 纬 -aminobutyric acid, glutamate receptors, and their receptors, in stress response or anxiety, depression, etc. In recent years, it has been found that TSPO may be a potential target for the treatment of affective mental disorders, especially anxiety disorders. Overexpression of 18kDa translocation protein TSPO in dentate gyrus of sea horse, The effect of depressive behavior provides experimental evidence for TSPO to become a potential new target for drugs. In this study, the adult male ICR mice were microinjected with lentivirus carrying TSPO gene in dentate gyrus, which resulted in overexpression of TSPO. The accuracy of fluorescent protein expression and the expression level of TSPO protein were observed after injecting lentivirus into the dentate gyrus of mice. On the 18th and 21st days, the despairing model of suspending tail and forced swimming in mice was used to detect depressive behavior, and the TSPO protein level and tetrahydroprogesterone content in the 3mm tissue around the hippocampal injection pore were detected by Western Blot and ELISA, respectively. To explore the relationship between the expression of TSPO and anxiety and depression in dentate gyrus of the sea horse. The results are as follows: 12 days after lentivirus injection, the level of TSPO protein in hippocampus of the overexpression group was significantly higher than that of the control group, and 12 days after the injection, the expression of TSPO protein in the hippocampus of the control group was significantly higher than that of the control group. The green fluorescent protein in the dentate gyrus of the sea horse was obvious in the overexpression group, and 12 days after the microinjection of lentivirus in the dentate gyrus of the mice. Compared with the LV-NC group, the LV-TSPO group significantly increased the entry time of mice into the central area, significantly increased the number of times of entering the open arm and the percentage of the opening arm time of the mice, and significantly decreased the standing times of the mice (P0.05). The times and time of entering the box indicated that the over-expression of TSPO in the dentate gyrus of the sea horse showed significant anti-anxiety behavior in mice. Compared with LV-NC group, LV-TSPO group significantly reduced the immobility time of mice in tail suspension test and forced swimming test, suggesting that TSPO overexpression in dentate gyrus of sea horse showed significant antidepressant behavior. Compared with the negative control group, The expression of TSPO in the dentate gyrus of the sea horse significantly increased the expression of TSPO protein in the 3mm tissue around the hippocampal injection pore, and the content of tetrahydroprogesterone increased significantly. The expression of TSPO in the dentate gyrus of the mice was significantly anti-anxiety. The behavioral effect of antidepressant and the elevated level of tetrahydroprogesterone mediated by TSPO may be the important mechanism.
【学位授予单位】：河北北方学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749

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