丁苯酞对神经发生中BDNF通路的激活和神经元细胞增殖的促进作用

发布时间：2018-01-29 04:06

本文关键词： 丁苯酞海马成体神经发生脑源性神经营养因子　出处：《武汉大学》2016年博士论文　论文类型：学位论文

【摘要】：目的：成体神经发生,特别是海马的成体神经发生被普遍认为与学习记忆能力有紧密联系。而丁苯酞(L-3-n-butylphthalide, NBP)作为临床上一种常用的改善轻、中度缺血性脑卒中,改善认知作用的药物,其改善脑卒中患者认知作用的机制极可能与其对海马齿状回中成体神经发生有密切关系。我们拟采用大鼠双侧颈总动脉永久结扎(bilateral common carotid artery occlusion, two vessel occlusion,2VO)这一目前较为公认的慢性脑缺血导致认知功能障碍的动物模型,我们给大鼠以不同浓度梯度的丁苯酞进行灌胃,从行为学评价整体认知,影响神经发生及相关分子学水平观察海马区成体神经发生在丁苯酞的认知改善疗效中的作用,并探讨可能涉及的分子机制。方法：将实验动物随机分为六组：假手术组(Sham),双侧颈总动脉永久结扎组(2VO),2VO+丁苯酞干预组(以不同浓度30,60,120mg/kg丁苯酞干预,连续四周给予大鼠灌胃)以及2VO+丁苯酞(120mg/kg灌胃)+K252a组(腹腔注射,25μg/kg)。使用水迷宫测试评估大鼠空间记忆和学习能力。使用5-溴-2’-脱氧尿苷(5-bromo-2'-deoxyuridine, BrdU)标记法探究丁苯酞对大鼠海马齿状回神经前体细胞增殖分化能力的影响。使用RT-PCR和Western Blot水平分析丁苯酞刺激下大鼠海马组织中脑源性神经营养因子(brain derived neurophic factor, BDNF),细胞外调节蛋白激酶(extracellular regulated protein kinases, ERK)和环磷酸腺苷反应元件结合蛋白(cAMP response element binding protein, CERB)的表达水平以及磷酸化水平变化。结果：研究结果显示,丁苯酞可以改善慢性脑缺血大鼠的空间记忆与认知学习能力。同时丁苯酞可以促进成体神经前体细胞的增殖,但其对神经分化没有影响。此外,丁苯酞可以上调大鼠慢性脑缺血组织中BDNF, pERK和pCERB的表达水平,并呈浓度依赖性,这一作用可能被BDNF酪氨酸激酶受体B(Tyrosin-kinase receptor B, TrkB)信号通路抑制剂K252a所抑制。结论：丁苯酞可改善大鼠空间记忆与认知水平,这一作用与成体神经的发生密切相关,而其促进成体神经的发生可能和上调BDNF, pERK和pCERB表达水平相关。
[Abstract]:Objective: adult neurogenesis, especially in the hippocampus, is generally considered to be closely related to the ability of learning and memory, while butyrophthalide L-3-n-butylphthalide. NBPs are commonly used clinically to improve mild and moderate ischemic stroke and to improve cognitive function. The mechanism of improving the cognitive function of stroke patients may be closely related to the occurrence of adult nerve in dentate gyrus. We intend to use bilateral common carotid artery ligation (bilateral common carotid artery ligation) in rats. Bilateral common carotid artery occlusion. Two vessel inclusion (2VOO) is a widely accepted animal model of cognitive impairment caused by chronic cerebral ischemia. The rats were given butyphthalide with different concentration gradient to evaluate the overall cognition from behavior. Effect of neurogenesis and related molecular level to observe the role of adult nerve in the cognitive improvement of butyphthalide. Methods: the experimental animals were randomly divided into six groups: sham-operated group and bilateral common carotid artery permanent ligation group. 2VO-butyphthalide intervention group (300.60g / kg butyphthalide at different concentrations). Rats were given intragastric administration of 2VO butylphthalide 120mg / kg for four consecutive weeks and K252a group (intraperitoneal injection). 25 渭 g / kg 路Water maze test was used to evaluate the spatial memory and learning ability of rats. 5-bromo-2deoxyuridine was used to evaluate the spatial memory and learning ability of rats. BrdU). Effects of butylphthalide on proliferation and differentiation of neural precursor cells in dentate gyrus of rat were investigated by labeling. RT-PCR and Western. Analysis of brain-derived neurotrophic factor (BDNF) in rat hippocampal tissue stimulated by butylphthalide (Blot). Brain derived neurophic factor. BDNF, extracellular regulated protein kinases. Erk) and camp response element binding protein. Changes in CERBs expression and phosphorylation levels. Results: the results showed that. Butyphthalide can improve the spatial memory and cognitive learning ability of rats with chronic cerebral ischemia. Butylphthalide can promote the proliferation of adult neural precursor cells, but it has no effect on neural differentiation. Butyphthalide up-regulated the expression of BDNF, pERK and pCERB in rats with chronic cerebral ischemia in a concentration-dependent manner. This effect may be attributed to the receptor of BDNF tyrosine kinase, B#en0# Tyrosin-kinase receptor B. Conclusion: butyphthalide can improve the spatial memory and cognitive level of rats, which is closely related to the development of adult nerve. It may be related to the up-regulation of BDNF, pERK and pCERB expression.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R338

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