Cyclic Mechanical Stretch Stress Enhances BDNF and NT-3 immunoreactivity in Cultured Spinal Cord Neuronal, Glial cells

循环机械拉伸应力增强培养的脊髓神经元、胶质细胞中的 BDNF 和 NT-3 免疫反应性

基本信息

批准号：
16390435
负责人：
BABA Hisatoshi
金额：
$ 7.81万
依托单位：
University of Fukui
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390435/
关键词：
Mechanical stress Neurotrophic factor cultured Spinal cord cell

项目摘要

Study Design. A rat model designed to investigate the effects of applied cyclic stretch stress on neurotrophic factor synthesis in the rat spinal cord cells in vitro.Objective. To evaluate the neurotrophic response in vitro neuron and glia cell under mechanical stress using immunohistological analysis.Summary of Background Data. Recent studies conducted in animals with experimental spinal cord damage examined the function, source and dynamics of induction of endogenious neurotrophic factors including brain-derived neurotrophic factor (BDNF), neurotrophin (NT)-3. To our knowledge, there are no studies that examined biological and pathological responses to mechanical stress in cultured cells derived from spinal cord.Methods. The cultured spinal cord cells were isolated from 14 day Sprague-Dawley rat embryos. For providing mechanical stress to cells, FX-3000TM-Flexercell Strain Unit (Flexercell International Corp., McKeesport, PA, USA) was used. After starting application of stimulation, the cell morphology and immunoreactivity was observed over time (0, 1, 2, 6, 12 and 48 hours).Results. Decrease in the number of surviving neuron, thickening of cell membrane and extention of intercellular substance were observed after providing stress to cells for 6 hours or more. The number of microtubule-associated protein (MAP)-2 positive cells decreases with time, while the number of glial fibrillary acidic protein GFAP positive cells was comparatively kept against stress. For astrocytes, the number of anti-GFAP/BDNF and NT-3 antibody positive cells was comparatively kept with the lapse of stress time, but observation over time revealed a high immuno-reactivity of the remaining anti-GFAP positive cells.Conclusion. Our results indicate that reactive astrocytes increase the expression of neurotrophic factors to the mechanical stress, a possibility that reactive astrocytes against mechanical stress in cultured cells are involved in the prolongation of survival and mechanical repair.

研究设计。目的：建立体外循环拉伸应力对大鼠脊髓细胞神经营养因子合成影响的大鼠模型。目的。使用免疫组织学分析评估机械应力下体外神经元和神经胶质细胞的神经营养反应。背景数据摘要。最近在实验性脊髓损伤动物中进行的研究检查了内源性神经营养因子（包括脑源性神经营养因子（BDNF）、神经营养蛋白（NT）-3）诱导的功能、来源和动态。据我们所知，还没有研究检查来自脊髓的培养细胞对机械应力的生物和病理反应。方法。培养的脊髓细胞是从 14 天的 Sprague-Dawley 大鼠胚胎中分离出来的。为了向细胞提供机械应力，使用 FX-3000TM-Flexercell 应变装置（Flexercell International Corp.，McKeesport，PA，USA）。开始施加刺激后，随着时间的推移（0、1、2、6、12 和 48 小时）观察细胞形态和免疫反应性。结果。对细胞施加应激6小时或更长时间后，观察到存活神经元数量减少、细胞膜增厚和细胞间质扩张。微管相关蛋白（MAP）-2阳性细胞的数量随着时间的推移而减少，而胶质纤维酸性蛋白GFAP阳性细胞的数量相对保持抵抗应激。对于星形胶质细胞，随着应激时间的推移，抗GFAP/BDNF和NT-3抗体阳性细胞的数量相对保持不变，但随着时间的推移观察发现剩余的抗GFAP阳性细胞具有高免疫反应性。结论。我们的结果表明，反应性星形胶质细胞增加了针对机械应力的神经营养因子的表达，这可能是培养细胞中针对机械应力的反应性星形胶质细胞参与了存活的延长和机械修复。