Acute Phase Protein Effects in Cellular Mechanisms of Microvascular Endothelium

急性期蛋白对微血管内皮细胞机制的影响

• 批准号: 7459156
• 负责人: Karen S Mark
• 金额: $ 9.24万
• 依托单位: UNIVERSITY OF MISSOURI KANSAS CITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-07 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7459156
• 关键词: Actins; Acute-Phase Proteins; Affect; American; Apical; Biological Markers; Blood; Blood - brain barrier anatomy; Blood Circulation; Brain; C-reactive protein; Cardiovascular Diseases; Cardiovascular system; Cell Line; Cells; Cerebrum; Chronic; Clinical; Communication; Complex; Culture Media; Development; Diabetes Mellitus; Diffusion; Disease; Disruption; Edema; Endothelial Cells; Endothelium; Enzyme-Linked Immunosorbent Assay; Exposure to; Foundations; Functional disorder; Future; Genetic Transcription; Goals; Haptoglobins; Health Care Costs; Homeostasis; Human; Hypoxia; Immune; Immune system; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Interleukin-1; Interleukin-10; Interleukin-4; Interleukin-6; Investigation; Ischemia; Lead; Leukocytes; Link; Little' s Disease; Measures; Mediator of activation protein; Modeling; Molecular; Molecular Weight; Monitor; Neurologic Dysfunctions; Neuropathy; Pathway interactions; Patients; Peripheral; Permeability; Pharmacological Treatment; Plague; Play; Polymerase Chain Reaction; Preventive; Process; Protein C; Proteins; Public Health; Quality of life; RNA; Rattus; Research; Research Proposals; Risk; Role; Serum amyloid A protein; Side; Signal Pathway; Signal Transduction; Stress; Stroke; Structural Protein; TNF gene; Testing; Tight Junctions; Time; Umbilical vein; Western Blotting; Work; activating transcription factor; base; brain cell; chronic pain; claudin-1 protein; clinically relevant; concept; cytokine; fighting; improved; insight; monolayer; neurovascular unit; occludin; programs; protein expression; research study; response; therapeutic target; transcription factor

DESCRIPTION (provided by applicant): Over the last decade, it has become apparent that dysregulation of the immune system is related to several serious diseases, including include diabetes, cardiovascular diseases, and chronic inflammation. Millions of Americans are affected by these diseases and results in increased healthcare costs. The imbalance of the immune/inflammatory responses in these diseases ultimately leads to cellular damage and pathophysiologic dysfunction like neuropathy and ischemia. While clinicians are working to identify key biomarkers to use in determining or monitoring the seriousness of these diseases, little has been done to identify the direct effects that inflammatory biomarkers have on the common target of the microvasculature. The release of acute phase proteins (APP), including haptoglobin, serum amyloid A, and C-reactive protein, occurs in many of these diseases, yet the underlying cellular impact of these inflammatory biomarkers is not well understood. Our hypothesis is that inflammatory mediators, such as APPs, act directly on the endothelial cells of the microvasculature, activating signaling pathways and cellular mechanisms that alter the normal endothelial barrier function. This proposal focuses on examining changes in the microvasculature following exposure to APPs to gain a better understanding of the underlying mechanisms that lead to plague development and disrupted endothelial transport. Using an in vitro BBB model (primary isolated rat brain microvessel endothelial cells, RBMEC) and a peripheral microvascular endothelial model (human umbilical vein endothelial cell line, HUVEC) we will examine the cellular mechanisms that are likely involved in structural and molecular changes of the microvascular endothelial barrier in response to APPs. Functional (BBB permeability), structural (expression of tight junction proteins) and molecular (release of related inflammatory cytokines) assessments will be carried out to establish a link between clinical findings of elevated levels of C-reactive protein, haptoglobin, etc. and the pathophysiological changes observed in the microvasculature of patients from afore mentioned diseases. The results from this study will provide the proof of concept for further investigation into clarifying cellular signaling pathways that are activated by these inflammatory biomarkers as well as identify ideal pharmacological treatments that will reduce the risks and improve the quality of life in patients with these diseases. The focus of this research proposal is to enhance our understanding of cellular mechanisms in the microvascular endothelium which respond to inflammatory mediators, especially acute phase proteins. The results of this study will provide invaluable information regarding the effects that acute phase proteins (i.e., C-reactive protein, haptoglobin, and serum amyloid A) stimulate in the microvascular endothelial barrier including permeability and related structural proteins that regulate these barriers. PUBLIC HEALTH RELEVANCE: The clinical impact of these studies will provide targets for therapeutic treatment as well as preventive measures for those patients with cardiovascular and inflammatory-related diseases.

描述（由申请人提供）：在过去的十年中，免疫系统的失调与几种严重疾病有关，包括糖尿病、心血管疾病和慢性炎症，这一点已经变得很明显。数百万美国人受到这些疾病的影响，导致医疗费用增加。这些疾病中免疫/炎症反应的失衡最终导致细胞损伤和病理生理功能障碍，如神经病变和缺血。虽然临床医生正在努力确定用于确定或监测这些疾病的严重性的关键生物标志物，但几乎没有做过什么来确定炎症生物标志物对微血管系统的共同靶点的直接影响。急性时相蛋白（APP），包括触珠蛋白，血清淀粉样蛋白A和C-反应蛋白的释放，发生在许多这些疾病中，但这些炎症生物标志物的潜在细胞影响还没有得到很好的理解。我们的假设是，炎症介质，如APP，直接作用于微血管内皮细胞，激活信号通路和细胞机制，改变正常的内皮屏障功能。该提案的重点是检查暴露于APP后微血管系统的变化，以更好地了解导致斑块发展和内皮运输中断的潜在机制。使用体外BBB模型（原代分离的大鼠脑微血管内皮细胞，RBMEC）和外周微血管内皮细胞模型（人脐静脉内皮细胞系，HUVEC），我们将研究可能参与微血管内皮屏障的结构和分子变化的细胞机制，以响应APP。将进行功能（BBB通透性）、结构（紧密连接蛋白的表达）和分子（相关炎性细胞因子的释放）评估，以建立C反应蛋白、触珠蛋白等水平升高的临床结果与上述疾病患者微血管中观察到的病理生理变化之间的联系。这项研究的结果将为进一步研究澄清由这些炎症生物标志物激活的细胞信号传导途径提供概念证据，并确定理想的药物治疗方法，以降低这些疾病患者的风险并改善其生活质量。这项研究的重点是加强我们对微血管内皮细胞机制的理解，这些机制对炎症介质，特别是急性时相蛋白做出反应。这项研究的结果将提供关于急性期蛋白（即，C-反应蛋白、触珠蛋白和血清淀粉样蛋白A）刺激微血管内皮屏障，包括渗透性和调节这些屏障的相关结构蛋白。公共卫生相关性：这些研究的临床影响将为心血管和炎症相关疾病的患者提供治疗和预防措施的靶点。