间脑外源组胺抑制斑马鱼脊髓损伤后修复的作用研究
发布时间：2018-08-10 22:39
【摘要】：以往关于脊髓损伤的研究,不管是以实验鼠或斑马鱼等为模式动物开展的基础性研究,还是临床上对于治疗脊髓损伤的新方法、新药物的尝试,基本都只关注受损的脊髓本身。本课题中,我们不再局限于受损的脊髓组织,而是从脑内的单胺类神经递质组胺(histamine,HA)着手,将同是中枢神经系统的脑组织同脊髓损伤联系起来,以期探究脑源性信号对下游脊髓组织修复的影响,发现影响脊髓损伤修复的新机制。作为一种来源于脑组织并下行至脊髓的单胺能神经递质,组胺在脊髓损伤修复中作用的报道较少;而已有研究表明脑内单胺能神经递质信号的多巴胺(dopamine,DA)和5-羟色胺(5-hydroxytryptamine,5-HT)具有重要的运动调节功能,对斑马鱼脊髓损伤后的修复有积极作用。我们采用成年斑马鱼,首先向其间脑内注射组胺,其后进行脊髓损伤手术,建立脑源性组胺水平升高的脊髓损伤斑马鱼模型,向对照组斑马鱼注射生理盐水并进行脊髓损伤。行为学分析的结果显示,间脑内组胺注射可以抑制脊髓损伤斑马鱼运动功能的恢复。免疫荧光染色结果显示,在脊髓损伤后的不同时间点,间脑组胺注射对损伤位点神经胶质细胞产生影响:损伤后第3天,脊髓损伤位点上段的小胶质细胞的荧光强度增强,第7天恢复至对照组水平;而对于放射状胶质细胞,是在损伤后第7天促进其增殖,在第11天恢复至对照组水平,但是,在损伤后第11天,损伤位点上段放射状胶质细胞呈现出不利于损伤后修复的多极星状形态。此外,组胺注射还加剧了下行到脊髓损伤位点的酪氨酸羟化酶阳性和5-羟色胺阳性神经纤维的崩解。实时定量聚合酶链式反应(qRT-PCR)的结果表明,斑马鱼间脑组胺注射使损伤位点上段的脑源性神经营养因子(BDNF)在损伤后第1天和第3天的表达减少;使胰岛素样生长因子1(IGF-1)在损伤后第3天的表达下调。以上实验结果表明,斑马鱼间脑组胺注射可以抑制成年斑马鱼脊髓损伤后运动功能的恢复,其机制可能是通过调节损伤位点的神经营养及免疫微环境,以及促进从脑组织下行到脊髓的其他单胺能神经纤维的崩解。本课题以脑源性信号—组胺为研究对象,探究了其对下游脊髓组织修复的影响,为发现脊髓损伤修复的新机制提供了独特思路。
[Abstract]:Previous studies on spinal cord injury, whether based on experimental rats or zebrafish as model animals, or clinical treatment of new methods of spinal cord injury, new drugs, basically only focus on the injured spinal cord itself. In this study, we no longer confine ourselves to the injured spinal cord, but start with the monoamine neurotransmitter histamine HA in the brain, linking the same brain tissue of the central nervous system to the spinal cord injury. The purpose of this study was to explore the effects of brain-derived signals on the repair of the downstream spinal cord, and to find out the new mechanism that affects the repair of spinal cord injury. As a monoaminergic neurotransmitter derived from brain tissue and descending to the spinal cord, histamine plays an important role in the repair of spinal cord injury. Some studies have shown that dopamine DA and 5-hydroxytryptamine 5-HT in the brain of monoaminergic neurotransmitter signal play an important role in motor regulation and play a positive role in the repair of spinal cord injury in zebrafish. In adult zebrafish, histamine was injected into the brain at first, then spinal cord injury was performed. The model of spinal cord injury was established with elevated histamine level. The normal saline was injected into the control group and spinal cord injury was carried out. Behavioral analysis showed that diencephalon histamine injection inhibited the recovery of motor function in spinal cord injury zebrafish. The results of immunofluorescence staining showed that at different time points after spinal cord injury, diencephalon histamine injection had an effect on the glial cells at the injured site: the fluorescence intensity of the microglia in the upper segment of the spinal cord injury site increased on the 3rd day after injury. On the 7th day, the radiative glial cells increased their proliferation on the 7th day after injury and recovered to the control level on the 11th day, but on the 11th day after the injury, the radiative glial cells returned to the level of the control group, but on the 11th day after the injury, The radial glial cells in the upper segment of injury site showed multipolar stellate morphology which was not conducive to repair after injury. In addition, histamine injection increased the disintegration of tyrosine hydroxylase positive and serotonin positive nerve fibers descending to the site of spinal cord injury. The results of real-time quantitative polymerase chain reaction (qRT-PCR) showed that diencephalon histamine injection decreased the expression of brain-derived neurotrophic factor (BDNF) in the upper part of the injury site on the 1st and 3rd day after injury. The expression of insulin-like growth factor 1 (IGF-1) was down-regulated on the 3rd day after injury. These results suggest that diencephalon histamine injection can inhibit the recovery of motor function after spinal cord injury in adult zebrafish, and its mechanism may be by regulating the neurotrophic and immune microenvironment of the injured site. And promote the disintegration of other monoaminergic nerve fibers descending from brain tissue to spinal cord. In this study, the effects of brain-derived signal histamine on the repair of the downstream spinal cord were investigated, which provided a unique idea for discovering the new mechanism of spinal cord injury repair.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R651.2
本文编号：2176433
[Abstract]:Previous studies on spinal cord injury, whether based on experimental rats or zebrafish as model animals, or clinical treatment of new methods of spinal cord injury, new drugs, basically only focus on the injured spinal cord itself. In this study, we no longer confine ourselves to the injured spinal cord, but start with the monoamine neurotransmitter histamine HA in the brain, linking the same brain tissue of the central nervous system to the spinal cord injury. The purpose of this study was to explore the effects of brain-derived signals on the repair of the downstream spinal cord, and to find out the new mechanism that affects the repair of spinal cord injury. As a monoaminergic neurotransmitter derived from brain tissue and descending to the spinal cord, histamine plays an important role in the repair of spinal cord injury. Some studies have shown that dopamine DA and 5-hydroxytryptamine 5-HT in the brain of monoaminergic neurotransmitter signal play an important role in motor regulation and play a positive role in the repair of spinal cord injury in zebrafish. In adult zebrafish, histamine was injected into the brain at first, then spinal cord injury was performed. The model of spinal cord injury was established with elevated histamine level. The normal saline was injected into the control group and spinal cord injury was carried out. Behavioral analysis showed that diencephalon histamine injection inhibited the recovery of motor function in spinal cord injury zebrafish. The results of immunofluorescence staining showed that at different time points after spinal cord injury, diencephalon histamine injection had an effect on the glial cells at the injured site: the fluorescence intensity of the microglia in the upper segment of the spinal cord injury site increased on the 3rd day after injury. On the 7th day, the radiative glial cells increased their proliferation on the 7th day after injury and recovered to the control level on the 11th day, but on the 11th day after the injury, the radiative glial cells returned to the level of the control group, but on the 11th day after the injury, The radial glial cells in the upper segment of injury site showed multipolar stellate morphology which was not conducive to repair after injury. In addition, histamine injection increased the disintegration of tyrosine hydroxylase positive and serotonin positive nerve fibers descending to the site of spinal cord injury. The results of real-time quantitative polymerase chain reaction (qRT-PCR) showed that diencephalon histamine injection decreased the expression of brain-derived neurotrophic factor (BDNF) in the upper part of the injury site on the 1st and 3rd day after injury. The expression of insulin-like growth factor 1 (IGF-1) was down-regulated on the 3rd day after injury. These results suggest that diencephalon histamine injection can inhibit the recovery of motor function after spinal cord injury in adult zebrafish, and its mechanism may be by regulating the neurotrophic and immune microenvironment of the injured site. And promote the disintegration of other monoaminergic nerve fibers descending from brain tissue to spinal cord. In this study, the effects of brain-derived signal histamine on the repair of the downstream spinal cord were investigated, which provided a unique idea for discovering the new mechanism of spinal cord injury repair.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R651.2
【参考文献】
相关期刊论文 前1条
1 Melitta Schachner;;A Surgery Protocol for Adult Zebrafish Spinal Cord Injury[J];遗传学报;2012年09期
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