COMPLEMENT AND INFLAMMATORY FACTORS IN AD PATHOGENESIS

描述：(改编自申请者摘要)：阿尔茨海默病(AD) 是一种常见的痴呆症或认知能力丧失，这与 脑组织变性。这项建议直接集中在 确定AD大脑中的补体激活是否触发了一条通路 导致神经病理和痴呆症。对这一事件的调查 发生的生化反应(C‘活化)之间的相互关系 以及由这些生化反应引发的细胞反应 反应可能导致确定可以 随后成为阿尔茨海默病治疗干预的靶点 病人。这一提议将检验这样一种假设，即 经典补体途径，同时导致细胞裂解破坏 在大脑内，也会触发局部炎症反应，这可能 导致神经变性和随后的认知功能障碍 阿尔茨海默氏症。大量的免疫化学证据表明， 补体在阿尔茨海默病脑中的作用已有报道。为了进一步评估体内的 这些事件的意义，各种定义的合成b-淀粉样蛋白 用于代表体内多肽的多肽将通过一种 灵敏的C4消耗测定，用于它们的相对激活能力 互补性。结果将与这些人的能力相关联 形成纤维的多肽(电子显微镜)以及 这些多肽之间的二级结构(圆二色)。在……里面 此外，已知的其他蛋白质对老年性痴呆的影响 此激活上的斑块(聚集素和α-抗凝乳杆菌胰蛋白酶)将 评估过了。在第二系列调查中，我们将调查 治疗前后小胶质细胞、星形胶质细胞和神经元的激活 补体激活片段C5a和/或 促炎细胞因子。产生超氧化物，触发 吞噬作用，蛋白水解酶的分泌，细胞因子和 补体蛋白将被确定。B-淀粉样蛋白的能力，或 B-淀粉样蛋白与补体起始蛋白C1q复合诱导 这些炎症活性也将进行测试，最初使用的是体外实验 在可能的情况下进行原代人类细胞培养，或在可能的情况下进行原代大鼠细胞培养 必要的和/或适当的。这些实验的结果将澄清 补体介导炎症的调节因子及其作用 神经退行性变和认知功能障碍的发生机制 在公元后。此外，这里的发现应该会增加对 中枢神经系统其他补体介导的病理改变。最后，中国的发展 这里描述的体外模型将允许测试潜在的药物 可能会调节有害活动，以减缓或阻止进展 这种疾病。

DESCRIPTION: (Adapted from Applicant's Abstract): Alzheimer's disease (AD) is a common dementia or loss of cognitive abilities, which is linked to the degeneration of brain tissue. This proposal is directly focussed on determining whether complement activation in AD brain triggers a pathway leading to neuropathology and dementia. Investigation of the interrelationships between biochemical reactions (C' activation) that occur and the cellular responses triggered as a result of those biochemical reactions could lead to identification of regulatory points that can subsequently be targeted for therapeutic intervention for Alzheimer disease patients. This proposal will test the hypothesis that the activation of the classical complement pathway, while resulting in lytic destruction of cells within the brain, also triggers a local inflammatory response which may contribute to the neurodegeneration and subsequent cognitive dysfunction in Alzheimer's disease. Abundant immunochemical evidence of the activation of complement in AD brain has been reported. To further assess the in vivo significance of these events, a variety of defined synthetic b-amyloid peptides, designed to represent the in vivo peptides will be assayed, via a sensitive C4 consumption assay, for their relative ability to activate complement. The results will be correlated with the ability of these peptides to form fibrils (electron microscopy) as well as variations in the secondary structure (circular dichroism) between these peptides. In addition, the effect of other proteins known to be present in the AD senile plaque (clusterin and a-antichymotrypsin ) on this activation will be assessed. In a second series of investigations, we will investigate the activation of microglia, astrocytes and neurons, before and after stimulation with the complement activation fragment C5a and/or proinflammatory cytokines. Generation of superoxide, triggering of phagocytosis, secretion of proteases, and production of cytokines and complement proteins will be determined. The ability of b-amyloid, or b-amyloid complexed with the complement initiation protein C1q to induce these inflammatory activities will also be tested, initially using in vitro primary human cell cultures when possible or primary rat cell cultures when necessary and/or appropriate. The results of these experiments will clarify the factors regulating, and the role of, complement-mediated inflammatory mechanisms in the development of neurodegeneration and cognitive dysfunction in AD. In addition, findings here should increase the understanding of other complement-mediated pathology in the CNS. Finally, the development of the in vitro model described here will allow testing of potential drugs that may modulate deleterious activities in an effort to slow or stop progression of this disease.