ENDOTHELIAL DYSFUNCTION: FORMATION OF MICROPARTICLES

内皮功能障碍：微粒的形成

• 批准号: 6942957
• 负责人: SERGEY BRODSKY
• 金额: $ 1.1万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2005-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6942957
• 关键词: biomarker; blood coagulation; blood coagulation disorders; blood vessel disorder; diabetes mellitus; flow cytometry; genetically modified animals; glucose; immunomagnetic separation; intermolecular interaction; laboratory mouse; laboratory rat; nitric oxide; particle; pathologic process; plasminogen activator inhibitors; protein binding; tissue /cell culture; vascular endothelium; vasoconstriction; western blottings

DESCRIPTION (provided by applicant): The proposed project should enhance candidate's previous experience in vascular biology and diabetes and develop the candidate into an independent researcher. While Pl's experience with in vivo and in vitro experimental techniques will help to conduct the presented project, its completion will significantly enrich candidate's knowledge in cell biology and physiology as well. Endothelial cell dysfunction (ECD) is a common precursor and denominator of various pathologic conditions, including stroke, hypertension, myocardial infarction and diabetes mellitus. The established hallmarks of endothelial cell dysfunction are a deficiency in the production or bioavailability of nitric oxide (NO), an increased level of plasminogen activator inhibitor-1 (PAl-1), and a presence of a pro-coagulant state. We have previously demonstrated that elevated level of PAl-1 results in a decreased production of NO by cultured endothelial cells, as well as an increased production of microparticies stained with Annexin V - evidence for the expression of anionic phospholipids. This was accompanied by the accelerated generation of thrombin by microparticles. PAl-1 knockout mice presented with a significantly decreased number of circulating microparticles and a marked increase in the number of microparticles after PAl-1 injection. Combined with recent findings of elevated levels of circulating endothelial microparticles in patients with acute coronary syndrome and diabetes, our data suggest that a) microparticles may be a consequence of ECD and contribute to a procoagulant state accompanying these diseases and b) raise the question whether increased levels of circulating microparticles may contribute to the development and maintenance of endothelial cell dysfunction. Based on these findings we hypothesize that: 1) increased microparticle formation by stressed endothelial cells is not only a novel marker of, but also a contributor to endothelial dysfunction and 2) elevated level of PAl-1 (a recognized marker of endothelial dysfunction) leads to increased formation of microparticles with pro-coagulant properties. The current proposal seeks to 1) establish a novel pathophysiolgical mechanism of endothelial dysfunction - formation of microparticles by stressed endothelial cells; 2) obtain information on the population of circulating microparticles produced by endothelial cells in diabetes mellitus; 3) to investigate possible links between identified markers of endothelial dysfunction, namely elevated levels of PAl-1 and increased coagulant activity. To accomplish these goals, microparticle formation will be studied using fluorescence-activated cell sorting in vitro (in endothelial cells stressed with high glucose or PAl-1) and in vivo (in Zucker diabetic rats). The link between PAl-1 and increased coagulant activity will be studied in vivo using PAl-1 knockout mice and in vitro using endothelial cell cultures. The proposed studies may provide novel information on the contribution of microparticles to the pathophysiology of endothelial cell dysfunction and coagulation abnormalities.

描述（由申请人提供）： 拟议的项目应增强候选人以前在血管生物学和糖尿病方面的经验，并将候选人发展为独立的研究人员。尽管PL在体内和体外实验技术方面的经验将有助于进行提出的项目，但其完成也将大大丰富候选人在细胞生物学和生理学方面的知识。 内皮细胞功能障碍（ECD）是各种病理状况的常见前体和分母，包括中风，高血压，心肌梗塞和糖尿病。内皮细胞功能障碍的已建立标志是一氧化氮（NO）的生产或生物利用度缺乏，纤溶酶原激活剂抑制剂1（PAL-1）的水平升高以及促凝状态的存在。我们先前已经证明，PAL -1的水平升高会导致培养的内皮细胞的NO产生降低，并增加了用膜联蛋白V染色的微观产生的增加 - 表达阴离子磷脂的证据。这伴随着微粒加速产生凝血酶。 PAL-1基因敲除小鼠的循环微粒数量明显减少，而PAL-1注射后的微粒数量显着增加。结合最近发现急性冠状动脉综合征和糖尿病患者循环内皮微粒升高水平的发现，我们的数据表明，a）微粒可能是ECD的结果，并且有助于伴随这些疾病的促进状态并伴随这些疾病并提高循环小颗粒的水平，以促进循环量的发展和维持的循环症状水平。根据这些发现，我们假设：1）由压力的内皮细胞形成的微粒形成不仅是新颖的标志物，而且是内皮功能障碍的贡献者，而2）PAL-1水平升高（公认的内皮功能障碍的标志物）导致具有pro型物质的微型分会形成增加。当前的提案寻求1）建立一种新型的病理生理学机制，其内皮功能障碍 - 由压力内皮细胞形成微粒； 2）获取有关糖尿病内皮细胞产生的循环微粒群体的信息； 3）研究内皮功能障碍鉴定的标记物之间的可能联系，即PAL-1水平升高和凝血活性增加。为了实现这些目标，将使用体外荧光激活的细胞分选（在高葡萄糖或PAL-1强调的内皮细胞中）和体内（在Zucker糖尿病大鼠中）研究微粒形成。使用PAL-1基因敲除小鼠和使用内皮细胞培养物在体内研究PAL-1与增加的凝血活性之间的联系。拟议的研究可以提供有关微粒对内皮细胞功能障碍和凝结异常的病理生理学的贡献的新信息。

项目成果

Dynamics of circulating microparticles in liver transplant patients.

• 发表时间: 2008-09
• 期刊: Journal of gastrointestinal and liver diseases : JGLD
• 作者: S. Brodsky;M. Facciuto;D. Heydt;Jun Chen;H. Islam;M. Kajstura;G. Ramaswamy;M. Aguero-Rosenfeld

Glycated Collagen I (GC) impairs angiogenesis in vitro: a study using an innovative chamber for cell research.

糖化胶原 I (GC) 会损害体外血管生成：一项使用创新室进行细胞研究的研究。

• DOI: 10.1016/j.diabres.2006.10.003
• 发表时间: 2007
• 期刊: Diabetes research and clinical practice
• 影响因子: 5.1
• 作者: Brodsky,SergeyV;Merks,RoelandMH;Mendelev,Natalia;Goo,Cara;Chen,Jun
• 通讯作者: Chen,Jun