Amyloid-B peptide on endothelial adhesion and its related cellular pathways

淀粉样蛋白B肽对内皮粘附及其相关细胞通路的影响

基本信息

批准号：
7939748
负责人：
JAMES C LEE
金额：
$ 18.52万
依托单位：
UNIVERSITY OF MISSOURI-COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7939748
关键词：
Actins Address Adhesions Advanced Glycosylation End Products Affinity Alzheimer&apos s Disease Amyloid Amyloid beta-Protein Antibodies Atomic Force Microscopy Binding Biochemical Biochemical Process Biological Biomechanics Blood - brain barrier anatomy Blood Vessels Brain Carbohydrates Cell Adhesion Cell Adhesion Molecules Cell membrane Cell physiology Cell surface Cells Cerebrum Characteristics Curcumin Data Deposition Development Disease E-Selectin Endothelial Cells Endothelium Event Free Radical Scavengers Goals Human Immunofluorescence Microscopy In Vitro Inflammatory Intervention Investigation Light MAPK14 gene MAPK8 gene Measurement Measures Mechanics Methodology Microglia Microscopy Mitogen-Activated Protein Kinases Mosses NADPH Oxidase Oxidative Stress P-Selectin Pathology Pathway interactions Peptides Peripheral Phosphotransferases Probability Process Production Property Reaction Reactive Oxygen Species Research Role Selectins Senile Plaques Signal Pathway Superoxides Surface Techniques Testing Western Blotting cantilever immunoreactivity in vivo inhibitor/antagonist insight monocyte monolayer monomer novel novel therapeutic intervention peptide B polymerization prevent public health relevance receptor

项目摘要

DESCRIPTION (provided by applicant): The overall goal of this project is to test the hypothesis that amyloid beta-peptide (A¿) in different self-assembled forms, monomers, oligomers and fibrils, directly impacts cerebral endothelial functions because it alters 1) the expression of adhesion molecules (e.g. P- and E-selectins) and actin polymerization; 2) cellular mechanical properties; and the forces involved in adhesion; and 3) these alterations are resulted from the signaling pathway involving the binding of A¿ to receptor for advanced glycation end products (RAGE) to induce activations of NADPH oxidase and MAP kinases. Alteration of the cerebral microvascular endothelium is intimately associated with Alzheimer's Disease (AD). Increased deposition of A¿ in cerebral vasculature and the increased presence of monocytes in the vessel wall and of activated microglia are frequently observed in AD brains. Recent studies show that peripheral monocytes can migrate across the blood-brain barrier (BBB) and differentiate into microglia within the brain parenchyma. In vitro studies demonstrated that A¿ deposition at the endothelial cell layer enhances the transmigration of monocytes. Thus, increased transmigration of monocytes into brains is thought to drive the disease development towards exacerbation of the oxidative and inflammatory conditions characteristic of the AD brain. Consistent with our hypothesis, our preliminary data show that A¿ oligomer increases the immunoreactivity of P-selectin at the endothelial surface and actin polymerization within endothelial cells. Since the transmigration of monocytes across the BBB is both a mechanical and a biochemical process, the expression of adhesion molecules and mechanical properties of endothelial cells are the critical factors that require investigation. In this regard, the direct effects of A¿ on the expression of adhesion molecules and the mechanical properties of endothelial cell have yet to be fully elucidated. Therefore, our study on how A¿ alters the factors governing the adhesion between monocytes and endothelial surfaces will provide highly relevant and novel biophysical information about the mechanism of progression of AD and development of intervention strategy. To test our central hypothesis, we propose three specific aims: Aim 1: Determine the effects of A¿ monomers, oligomers and fibrils on the expression of P- and E- selectins and actin polymerization in immortalized cerebral endothelial cells (CECs). Aim 2: Measure the effects of A¿ monomers, oligomers and fibrils on the cell mechanical and adhesion properties of CECs. Aim 3: Determine the roles of RAGE, NADPH oxidase and MAP kinases (i.e. p38, ERK and JNK) in A2-induced alterations in endothelial adhesion. These aims will be accomplished by various biophysical and biochemical techniques including quantitative immunofluorescent microscopy, atomic force microscopy and various biochemical approaches. PUBLIC HEALTH RELEVANCE: Alteration of the cerebral microvascular endothelium is intimately associated with Alzheimer's Disease. We use atomic force microscopy to investigate the mechanism underlying the altered adhesion properties of endothelium induced by amyloid beta peptide. The mechanism is important in part of Alzheimer's disease development.

描述（由申请人提供）：该项目的总体目标是检验以下假设：不同自组装形式、单体、寡聚体和原纤维的淀粉样蛋白 β 肽 (A¿) 直接影响脑内皮功能，因为它改变了 1)粘附分子（例如 P-和 E-选择素）和肌动蛋白聚合的表达；以及粘附所涉及的力； 3）这些改变是由涉及A¿结合的信号通路引起的晚期糖基化终末产物 (RAGE) 受体诱导 NADPH 氧化酶和 MAP 激酶的激活脑微血管内皮细胞的改变与阿尔茨海默病 (AD) 的沉积增加密切相关。最近的研究表明，AD 大脑中经常观察到脑血管系统中单核细胞的存在增加以及激活的小胶质细胞的存在，外周单核细胞可以迁移穿过血脑屏障（BBB）并分化为大脑内的小胶质细胞。体外研究表明，A¿内皮细胞层的沉积增强了单核细胞的迁移，因此，单核细胞向大脑的迁移增加被认为会导致 AD 脑部氧化和炎症特征的恶化，这与我们的假设一致，我们的初步数据显示。那个A??寡聚物增加内皮表面 P-选择素的免疫反应性和内皮细胞内肌动蛋白的聚合作用。由于单核细胞跨 BBB 的迁移既是机械过程又是生化过程，因此粘附分子的表达和内皮细胞的机械特性至关重要。需要调查的因素在这方面，A¿的直接影响。粘附分子的表达和内皮细胞的机械特性的关系尚未完全阐明，因此，我们对A¿改变控制单核细胞和内皮表面之间粘附的因素将为 AD 进展机制和干预策略的制定提供高度相关和新颖的生物物理信息。为了检验我们的中心假设，我们提出了三个具体目标：目标 1：确定效果。的 A?单体、寡聚体和原纤维对永生化脑内皮细胞 (CEC) 中 P- 和 E- 选择素以及肌动蛋白聚合的表达目标 2：测量 A¿目标 3：确定 RAGE、NADPH 氧化酶和 MAP 激酶（即 p38、ERK 和 JNK）在 A2 诱导的内皮粘附变化中的作用。通过各种生物物理和生化技术完成，包括定量免疫荧光显微镜、原子力显微镜和各种生化技术接近。公共健康相关性：脑微血管内皮的改变与阿尔茨海默病密切相关，我们使用原子力显微镜来研究淀粉样蛋白肽诱导的内皮粘附特性改变的机制，该机制在阿尔茨海默病的发展中很重要。