STUDY OF THE MECHANISM OF NEURONAL CELL DAMAGE IN DEMENTIA FOR EARLY DIAGOSIS

痴呆症神经细胞损伤机制研究及早期诊断

基本信息

批准号：
18591303
负责人：
SOHMA Hitoshi
金额：
$ 2.55万
依托单位：
Sapporo Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

In dementia such as Alzheimer's disease (AD) morphological change due to neuronal cell apoptosis is observed in the brain. The cell damage in dementia has been reported to be caused by functional disorder of mitochondria and microsomes. Intracellular elevation of Ca^<2+> is involved in the pathophysiology of AD due to accumulation of peptide A. AD is often diagnosed at a moderately advanced stage. Early detection is required for early treatment. In this study we aimed to investigate molecular markers representing AD. We first analyzed secreted proteins from culture cells (human neuroblastoma NB-1 and rat glioma C6 cells), induced by Ca^<2+>-damage with A23187. Then, we analyzed the protein in blood plasma of dementia patients. As the blood-brain barrier strictly limits transport into the brain through both physical (tight junctions) and metabolic (enzymes) barriers and relatively hydrophobic materials tends to go through the barrier, we isolated the lipid liposome (PC : PS=9 : 1) binding proteins. Annexiss A5, A2 and fetuin were identified after separation on 2D-gel with LC-MS. As annexin A2 is highly expressed in blood cells, we focused on annexin A5. In the next step, blood plasma level of annexin A5 was measured using sandwich ELISA with AD, other dementia patients and controls. The level of plasma annexin A5 was 3.92±2.08 ng/ml in AD, while that in control was 0.87±0.21 ng/ml. The plasma annexin A5 level tended to increase with decreasing level of Hasegawa dementia scale (HDS-R) score. On the other hand, plasma annexin A5 level tended to be high (0.87±1.29 ng/ml) in vascular dementia patients, although not significantly higher than in controls. The physiological role of annexin A5 in the etiology of AD is next to be clarified.

在痴呆症中，例如在大脑中观察到神经元细胞凋亡引起的阿尔茨海默氏病（AD）形态学变化。据报道，痴呆症中的细胞损伤是由线粒体和微粒体功能障碍引起的。由于肽A的积累，Ca^<2+>的细胞内升高参与AD的病理生理学。AD通常在中等晚期的阶段被诊断出来。早期治疗需要尽早检测。在这项研究中，我们旨在研究代表AD的分子标记。我们首先分析了来自培养细胞（人类神经母细胞瘤NB-1和大鼠神经胶质瘤C6细胞）的分泌蛋白质，该蛋白质由Ca^<2+> - 用A23187损伤诱导。然后，我们分析了痴呆患者血浆中血浆中的蛋白质。由于血脑屏障严格限制了通过物理（紧密连接）和代谢（酶）屏障和相对疏水材料往往会穿过屏障，因此我们分离了脂质脂质体（PC：PS：PS = 9：1）结合蛋白。用LC-MS在2D凝胶上分离后，鉴定出Anexiss A5，A2和Fetuin。由于膜联蛋白A2在血细胞中高度表达，因此我们专注于膜联蛋白A5。下一步，使用带有AD，其他痴呆症患者和对照组的三明治ELISA测量膜联蛋白A5的血浆水平。在AD中，血浆附着素A5的水平为3.92±2.08 ng/ml，而对照中的水平为0.87±0.21 ng/ml。血浆膜蛋白A5水平往往会随着谷川痴呆量表（HDS-R）评分的降低而增加。另一方面，血浆膜联蛋白A5水平在血管痴呆患者中倾向于高（0.87±1.29 ng/ml），尽管并不明显高于对照组。接下来要澄清膜联蛋白A5在AD病因中的身体作用。