BDNF-GABA能神经功能障碍与老年焦虑症关系的研究

发布时间：2018-02-16 01:37

本文关键词： 老年焦虑症脑源性神经营养因子 γ-氨基丁酸能神经神经可塑性　出处：《云南中医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的:近年来,老年焦虑症发病率与日俱增,成为严重影响老年人健康的疾病。γ-氨基丁酸(γ-aminobutyric acid,GABA)能神经是调节情绪的主要神经,其功能障碍可导致焦虑行为的发生。脑源性神经营养因子(brain derived neurotrophic factor,BDNF)参与GABA能神经可塑性的调节,并具有改善焦虑相关行为的作用,其水平随着年龄的增长而减少。因此,我们提出假设老年人随BDNF的减少,导致GABA能神经功能障碍,焦虑易感性增加,补充外源性BDNF可通过调节GABA能神经可塑性发挥抗焦虑作用。本实验将探讨BDNF与GABA能神经功能与老年焦虑症的相关性,以及通过补充BDNF改善GABA能神经可塑性治疗老年焦虑症的可行性与有效性,为开发老年焦虑症的治疗寻找新的药物靶点。方法:1.第一部分BDNF缺乏-GABA能神经功能障碍-焦虑行为相关性的研究实验分3组:3月龄阴性对照组(3Mon)、12月龄阴性对照组(12Mon)、12月龄BDNF-LVsh RNA组(12Mon/KD)。通过齿状回注射BDNF-LVsh RNA敲低老龄小鼠内源性BDNF的表达,以高架十字迷宫法(EPM)研究小鼠焦虑程度,以ELISA检测小鼠海马、皮层的GABA和BDNF水平。最后进行行为学和生化指标的相关性分析,研究BDNF缺乏对老龄小鼠GABA能神经功能及焦虑行为的影响。2.第二部分外源性BDNF抗老年焦虑的作用与GABA能神经可塑性的相关性研究实验分4组:3月龄对照组(3Mon)、12月龄对照组(12Mon)、12月龄BDNF组(12Mon/BDNF)、12月龄氟西汀组(12Mon/FXT)。通过给予老龄小鼠外源性BDNF,以EPM研究小鼠焦虑程度;以跳台法研究小鼠学习记忆能力。以ELISA检测小鼠海马、皮层的GABA和BDNF水平,利用Western blot法检测GABAA-Rα1、α2和α5亚基的表达。最后进行行为学和生化指标的相关性分析,研究外源性BDNF对12月老龄小鼠GABA神经可塑性及焦虑行为的改善作用。结果:1.BDNF缺乏-GABA能神经功能障碍-焦虑行为相关性的研究1.1在EPM行为检测中,与3Mon组相比,12Mon组和12Mon/KD组进入开臂次数的百分比(OE%)有降低趋势。与12Mon组相比,12Mon/KD组进入开臂时间的百分比(OT%)有降低趋势。1.2与3Mon组相比,12Mon/KD组的皮层BDNF水平明显降低,差异有统计学意义(P0.05);12Mon组和12Mon/KD组的海马BDNF水平明显降低,差异有统计学意义(P0.001;P0.01)。与12Mon组相比,12Mon/KD组海马、皮层的GABA水平有降低趋势。1.3相关性分析结果表明,小鼠的海马、皮层中GABA水平、BDNF水平与OT%和OE%存在正相关性(P0.05);小鼠的海马、皮层中GABA水平和BDNF水平存在正相关性(P0.05)。2.外源性BDNF抗老年焦虑的作用与GABA能神经可塑性的相关性研究2.1在EPM行为检测中,与12Mon组相比,12Mon/BDNF组和12Mon/FXT组的OT%和OE%均明显升高,差异有统计学意义(P0.05;P0.01);3Mon组的OE%明显升高,差异有统计学意义(P0.01)。在跳台检测中,与12Mon组相比,第一天学习能力测试,12Mon/BDNF组的错误潜伏期明显升高,差异有统计学意义(P0.05);第二天记忆能力测试,12Mon/BDNF组的错误次数明显减少,错误潜伏期明显增长,差异有统计学意义(P0.01;P0.05)。2.2与12Mon组相比,12Mon/FXT组和12Mon/BDNF组的海马内BDNF水平和GABA水平明显升高,差异有统计学意义(P0.05;P0.001);皮层内BDNF水平和GABA水平有升高趋势。2.3与12Mon组相比,12Mon/BDNF组的海马、皮层GABAA-Rα2和α5亚基表达水平均明显升高,差异有统计学意义(P0.05)。与3Mon组相比,12Mon组的海马GABAA-Rα5亚基表达水平明显降低,差异有统计学意义(P0.01)。2.4小鼠海马、皮层中GABA水平和OT%存在正相关性(P0.05),小鼠海马中BDNF水平和OE%存在正相关性(P0.05);小鼠皮层中BDNF水平和OT%存在正相关性(P0.05)。小鼠海马、皮层中GABA水平和BDNF水平存在正相关性(P0.05)。小鼠海马、皮层中GABAA-Rα2和α5亚基表达水平与BDNF水平存在正相关性(P0.05)。结论:1.敲低内源性BDNF m RNA可增加老龄小鼠焦虑行为易感性,其作用机制与BDNF水平降低导致GABA能神经功能障碍有关。2.外源性BDNF可改善老龄小鼠的焦虑行为和学习记忆功能,其作用机制与改善脑内GABA能神经可塑性有关。3.BDNF可能是治疗老年焦虑症潜在药物靶点。
[Abstract]:Objective: in recent years, the incidence of anxiety disorder in the elderly grow with each passing day become a serious impact on the health of the elderly, the disease. GABA (gamma -aminobutyric acid, GABA) is the main nerve nerve regulating emotions, its dysfunction can lead to anxiety behavior. Brain derived neurotrophic factor (brain derived, neurotrophic factor, BDNF) participate in GABA can regulate neural plasticity, and can improve the anxiety related behavior, its level decreased with age. Therefore, we propose that the elderly with the decrease of BDNF, GABA can cause nerve dysfunction, anxiety and increased susceptibility to exogenous BDNF can be adjusted by the GABA can exert anti neural plasticity the role of anxiety. This experiment will explore BDNF and GABA correlation of nerve function and anxiety disorder in the elderly, and supplemented by BDNF GABA can improve neural plasticity in treating anxiety disorder in the elderly can Feasibility and effectiveness, for the development of treatment of anxiety disorder in the elderly to find new drug targets. Methods: part 1. BDNF lack of -GABA can experimental study on relativity between anxiety behavior - nerve dysfunction were divided into 3 groups: negative control group (3 month old 3Mon 12 month old), negative control group (12Mon), RNA group (12Mon/KD BDNF-LVsh 12 month old). Through the injection of BDNF-LVsh RNA expression in dentate gyrus of knockdown of endogenous BDNF in aged mice, elevated plus maze method (EPM) on mouse anxiety, detected by ELISA in mouse hippocampus cortex, GABA and BDNF levels. Finally, correlation analysis of behavior and biochemical effects of GABA and BDNF, the lack of research on aging mice GABA can the nerve function and anxiety behavior influence.2. second part of exogenous BDNF anti senile anxiety correlation study of neural plasticity were divided into 4 groups: 3 month old control group (3Mon), 12 month old in the control group (12Mon), December At the age of BDNF group (12Mon/BDNF) and fluoxetine group (12Mon/FXT). 12 month old by exogenous BDNF in aged mice, EPM mice in step-down test anxiety; Study on learning and memory ability of mice. ELISA mice were tested, cortex GABA and BDNF level detection using Western blot GABAA-R alpha 1, alpha 2 expression and the alpha 5 subunit. Finally, correlation analysis and biochemical behavior, improve the effect of exogenous BDNF on December aged mice GABA neural plasticity and anxiety behavior. Results: 1.BDNF -GABA lack of nerve dysfunction and anxiety for research on the relationship between the 1.1 in EPM detection, compared with 3Mon group, 12Mon the percentage of group and 12Mon/KD group of open arm entries (OE%) were decreased. Compared with 12Mon group, 12Mon/KD group the percentage of open arm time (OT%) decreased.1.2 compared with 3Mon group, 12Mon/KD group of cortical BDNF levels Decreased obviously, the difference was statistically significant (P0.05); 12Mon group and 12Mon/KD group, the BDNF level in the hippocampus decreased significantly, the difference was statistically significant (P0.001; P0.01). Compared with 12Mon group, 12Mon/KD group, hippocampus, cortex GABA levels had decreased.1.3 correlation analysis results showed that the mice hippocampus, the level of GABA in cortex the level of BDNF and OT% and OE% (P0.05); there is a positive correlation between the hippocampus of mice, GABA and BDNF level in the cortex (P0.05) there is a positive correlation between.2. and GABA effects of exogenous BDNF anti senile anxiety to neural plasticity related research in 2.1 EPM behavior detection, compared with 12Mon group, 12Mon/BDNF group group 12Mon/FXT and OT% and OE% were significantly increased, the difference was statistically significant (P0.05; P0.01); group 3Mon OE% increased significantly, the difference was statistically significant (P0.01). In the step-down test, compared with the 12Mon group, the first day of learning ability test, 12Mon/BDN F group the error latency was significantly increased, the difference was statistically significant (P0.05); the second day memory test, 12Mon/BDNF group significantly reduced the number of errors, error latency significantly increased, the difference was statistically significant (P0.01; P0.05).2.2 compared with 12Mon group, the levels of BDNF and GABA in hippocampus of 12Mon/FXT group and 12Mon/BDNF group obviously increased, the difference was statistically significant (P0.05; P0.001); the levels of BDNF and GABA in cerebral cortex increased.2.3 compared with 12Mon group, in 12Mon/BDNF group, GABAA-R in cerebral cortex of alpha 2 and alpha 5 subunit expression levels were significantly increased, the difference was statistically significant (P0.05). Compared with 3Mon group, 12Mon group hippocampal GABAA-R alpha 5 subunit expression levels were significantly lower, the difference was statistically significant (P0.01) in hippocampus of mice.2.4, GABA level and OT% cortex (P0.05), there is a positive correlation between the level of BDNF and OE% in the hippocampus of mice has a positive correlation (P0.05); The level of BDNF and OT% in the cortex. There is a positive correlation (P0.05) in hippocampus of mice, GABA and BDNF level in the cortex (P0.05). There was a positive correlation between the mouse hippocampus, cortex GABAA-R alpha 2 and alpha 5 subunit expression level and BDNF level has a positive correlation (P0.05). Conclusion: 1. knockdown of endogenous BDNF m RNA can increase the anxiety behavior in aging mice reduced susceptibility, the mechanism and the level of BDNF GABA can lead to anxiety behavior and learning memory function of neural dysfunction.2. exogenous BDNF can improve the aging mice, its mechanism and improvement of GABA in the brain can neural plasticity may be related to the.3.BDNF treatment of elderly patients with anxiety disorder of potential drug targets.

【学位授予单位】：云南中医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R749.72

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