Development of the Enzyme Immunoassay Systems for the Substances Involved in the Cause of Alzheimer's Disease

开发与阿尔茨海默氏病相关的物质的酶免疫测定系统

• 批准号: 63870036
• 负责人: HAYASHI Kyozo
• 金额: $ 6.66万
• 依托单位: Gifu Pharmaceutical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63870036/
• 关键词: Alzhermer's disease; cholinergic nerve cells; nerve growth factor (NGF); superoxide dismutase (SOD); enzyme immunoassay system (EIA); antibody; diagnosis; astoglial cells; 老人痴呆塩; 抗体; 診断; 神経成長促進因子; 線維芽細胞成長因子; アミロイド蛋白; 老人痴呆; 測定感度; 早期診断

Alzheimer's disease is a disease of unknown cause that is characterized by a progressive loss of memory and other cognitive functions, leading to a severe form of dementia and ultimately death. Neuropathologically, senile plaques and neurofibrillary tangles constitute the main features of Alzheimer's disease and it appears likely that they lead to the neuronal degeneration that characterizes the disease. One of the most consistent neuropathdlogical findings is the degeneration of magnocellular cholinergic nerve cells in the nucleus basalis of Meynert. Nerve growth factor (NGF) administration reduces the basal forebrain cholinergic neuron atrophy and the spatual memory impairment that are associated with normal aging in rats. Purpose of this research was thus to develop the highly sensitive enzyme immunoassay (EIA) systems for various neurotrophic factors involved in the cause of Alzhei mer's disease and to apply these developed EIA systems for the diagnosis of the Alzheimer's disease.M … More ajor results are summarized as follows; (1) We developed the purification procedures for many biologically active substances, which are concerned in the cause of the disease, including NGF, epidermal growth factor (EGF), superoxide dismutase (SOD), pletelet-derived growth factor (PDGF), etc. (2) By using the highly purified these substances, we prepared their monoclonal and polyclonal antibodies against rabbits of mice. (3) By using these antibodies prepared, we developed their highly sensitive, simple, and reliable two-site enzyme immunoassay systems. The minimum amounts of these substances detected by these enzyme immunoassay systems were approximately 10-15 pg/ml when a 100 pg aliquot of the sample was used. Good reproducibilities of within- and between-assay series and excellent recovery of exogenous substances from serum were observed. (4) We demonstrated that catecholamines increased the NGF content in medium conditioned by mouse L-M fibroblast cells and mouse astroglial cells. In this research, we demonstrated that catecholamines enhanced NGF synthesis of L-M cells and astroglial cells by increasing the cellular content of NGF MRNA and the affects of catecholamines were not mediated by adrenergic receptors. Coenzyme Q and vitamin K analogues also found to stimutate the NGF synthesis of astroglial cells. Less

阿尔茨海默病是一种病因不明的疾病，其特征是记忆和其他认知功能的进行性丧失，导致严重的痴呆症并最终死亡。神经病理学上，老年斑和神经纤维缠结构成阿尔茨海默病的主要特征，并且似乎它们可能导致作为疾病特征的神经元变性。最一致的神经病理学发现之一是Meynert基底核中大细胞胆碱能神经细胞的变性。神经生长因子（NGF）管理减少基底前脑胆碱能神经元萎缩和与大鼠正常衰老相关的记忆障碍。本研究的目的是建立一种高灵敏度的神经营养因子酶免疫分析系统，并将其应用于阿尔茨海默病的诊断。 ...更多信息 主要结果如下：（1）建立了与该病病因有关的多种生物活性物质的纯化方法，包括神经生长因子（NGF）、表皮生长因子（EGF）、超氧化物歧化酶（SOD）、血小板衍生生长因子（PDGF）等。（2）利用这些高纯度物质制备了抗兔或小鼠的单克隆抗体和多克隆抗体。(3)利用这些抗体，我们建立了高灵敏度、简便、可靠的双位点酶免疫分析系统。当使用100 pg等份样品时，这些酶免疫测定系统检测到的这些物质的最低量约为10-15 pg/ml。观察到试验内和试验间系列的良好重现性和从血清中回收外源性物质的良好回收率。(4)我们证明，在小鼠L-M成纤维细胞和小鼠星形胶质细胞的条件培养基中，儿茶酚胺增加NGF含量。在本研究中，我们证明了儿茶酚胺通过增加细胞内NGFmRNA的含量来促进L-M细胞和星形胶质细胞的NGF合成，并且儿茶酚胺的作用不是由肾上腺素能受体介导的。辅酶Q和维生素K类似物也被发现刺激星形胶质细胞的NGF合成。少

项目成果

林恭三: "神経成長促進因子(2)" 化学と薬学の教室. 34-39 (1989)

Kyozo Hayashi：“神经生长促进因子（2）”化学与药学系34-39（1989）。

林恭三: "新生化学実験講座 第11巻 神経生化学(日本生化学会編)"

林恭三：《新生物化学实验教程第11卷神经生物化学（日本生化学会编）》

Daisuke Kohda, Chie Kodama, Ryoichi Kase, Hiroshi Nomoto, Kyozo Hayashi, and Fuyuhiko Inagami: "A Comparative ^1H NMR study of mouse alpha(1-53) and beta(2-53) epidermal growth factors." Biochem.Int., 16, 647-654 (1988).

Daisuke Kohda、Chie Kodama、Ryoichi Kase、Hiroshi Nomoto、Kyozo Hayashi 和 Fuyuhiko Inagami：“小鼠 α(1-53) 和 β(2-53) 表皮生长因子的比较 ^1H NMR 研究。”

Yoshiko Furukawa, Nobuyuki Fukazawa, Yukio Miyama, Kyozo Hayashi, and Shoei Furukawa: "Stimulatory effect of 4-alkylcatechols and their diacetylated derivatives on the synthesis of nerve growth factor." Biochem.Pharmacol.

Yoshiko Furukawa、Nobuyuki Fukazawa、Yukio Miyama、Kyozo Hayashi 和 Shoei Furukawa：“4-烷基儿茶酚及其二乙酰化衍生物对神经生长因子合成的刺激作用。”

Ichizo Shinoda, Rie Takeuchi, Masayuki Kurobe, Shoei Furukawa, and Kyozo Hayashi: "Molecular nature of beta-nerve growth factor (NGF)-like immunoreactive substance(s) in mouse plasma." J.Clin.Biochem.Nutr., 5, 135-144 (1988).

Ichizo Shinoda、Rie Takeuchi、Masayuki Kurobe、Shoei Furukawa 和 Kyozo Hayashi：“小鼠血浆中 β 神经生长因子 (NGF) 样免疫反应物质的分子性质。”