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

阿尔茨海默病是一种病因不明的疾病，其特征是记忆力和其他认知功能的逐渐丧失，导致严重形式的痴呆，最终导致死亡。神经病理学上，老年斑和神经原纤维缠结构成阿尔茨海默病的主要特征，它们似乎很可能导致该疾病的特征神经元变性。最一致的神经病理学发现之一是Meynert基底核的大细胞胆碱能神经细胞变性。神经生长因子（NGF）可减轻大鼠基底前脑胆碱能神经元萎缩和与正常衰老相关的空间记忆障碍。因此，本研究的目的是开发用于阿尔茨海默病病因的各种神经营养因子的高灵敏度酶免疫测定（EIA）系统，并将这些已开发的EIA系统应用于阿尔茨海默病的诊断。更多的主要结果总结如下：(1)建立了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）等多种与疾病病因有关的生物活性物质的纯化方法。(2)利用高纯度的这些物质制备了抗兔/小鼠的单克隆和多克隆抗体。(3)利用所制备的抗体，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的样品时，这些酶免疫测定系统检测到的这些物质的最小量约为10-15 pg/ml。结果表明，该方法具有良好的重复性和血清中外源性物质的回收率。(4)在小鼠L-M成纤维细胞和小鼠星形胶质细胞培养基中，儿茶酚胺增加了NGF的含量。在本研究中，我们证明儿茶酚胺通过增加细胞中NGF MRNA的含量来促进L-M细胞和星形胶质细胞的NGF合成，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类似物也被发现可以刺激星形胶质细胞的NGF合成。少

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