Genistein and cAMP signaling in vascular endothelium

血管内皮细胞中的金雀异黄素和 cAMP 信号传导

• 批准号: 7140738
• 负责人: DONGMIN LIU
• 金额: $ 19.98万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7140738
• 关键词: Address; Adenylate Cyclase; Adhesions; Aging; American; Animals; Arteriosclerosis; Biochemical; Biological; Biological Assay; Blood Pressure; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Cell Proliferation; Cell membrane; Cells; Chronic; Condition; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cytoskeletal Modeling; Data; Depressed mood; Development; Disease; Dose; Elevation; Endothelial Cells; Estrogen Receptors; Event; Extravasation; Foundations; Functional disorder; GTP-Binding Proteins; Gases; Gene Expression; Genetic Transcription; Genistein; Genomics; Goals; Health; Health Benefit; Hypertension; Hypotension; Ingestion; Intervention; Isoflavones; Leukocytes; Mediating; Mediation; Messenger RNA; Methods; Microscope; Molecular; Morbidity - disease rate; Nitric Oxide; Nitric Oxide Synthase; Nutraceutical; Pathway interactions; Permeability; Physiological; Play; Polymerase Chain Reaction; Population; Preventive; Process; Production; Protein Biosynthesis; Protein Tyrosine Kinase; Proteins; Public Health; RNA; Range; Rattus; Research; Ribonucleases; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Smooth Muscle Myocytes; Soy Proteins; System; Testing; Therapeutic; Thrombin; Time; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelium; analog; base; cellular targeting; dietary supplements; human NOS3 protein; immunocytochemistry; monolayer; non-genomic; novel; promoter; protective effect; protein expression; soy; soy protein isolate; vasoactive agent

DESCRIPTION (provided by applicant): The long-range goal of this research is to develop novel vasoactive agents that may be of benefit in lowering the high level of cardiovascular morbidity in the aging American population. Recent studies have shown that ingestion of soy protein associated with isoflavones may have a protective role in vasculature. Genistein, the major soy isoflavone, has various biologgical actions. However, it is not clear whether the beneficial effect of soy protein is due to genistein or other components. In addition, the cellular or molecular mechanisms underlying the vessel protective effect of soy products remain unclear. We recently found that genistein directly acts on vascular dothelial cells (EC), leading to cAMP accumulation, nitric oxide synthase (eNOS) activation and expression. Further, We demonstrated that genistein induced a rapid decrease in systemic blood pressure (BP) in rats. The objective of this application, is therefore to determine how genistein triggers the cAMP signaling pathway and thereby influencing the cAMP-related vascular function. The four following questions will be addressed: 1). How does genistein elevate cAMP, activate eNOS, and consequently stimulate nitric oxide in EC? 2) Does genistein decrease BP in animals through activation of the cAMP signaling? 3). Does elevated cAMP by genistein also alter eNOS gene expression ? 4). Does genistein protect endothelial barrier function via the cAMP signaling? We will employ cellular, molecular and biochemical analysis to determine whether genistein elevates cAMP via activation of Gs-mediated adenylate cyclase in EC. Genistein-regulated eNOS gene expression will be evaluated by ribonuclease protection assay and Real-Time PCR. The effect of genistein on endothelial barrier function, as determined by cytoskeletal reorganization and barrier permeability, will be assessed using immunocytochemistry, confocal microscope and macromolecular leakage methods. Pharmacological and molecular intervention approaches will be used to detemine the involvement of a specific pathway in mediation of genistein action. The results from these studies will establish the mechanisms of the genistein action in vasculature and thereby provide foundation for further studies to determine the effects of genistein on vascular health and diseases such as hypertension. The demonstration of the health benefits of soy isoflavones in vasculature is expected to have significant effect on public health since isoflavones are widely used as a dietary supplements.

描述（由申请人提供）：本研究的长期目标是开发新型血管活性药物，可能有助于降低美国老龄化人口中高水平的心血管发病率。最近的研究表明，摄入与异黄酮相关的大豆蛋白可能对脉管系统具有保护作用。金雀异黄酮是主要的大豆异黄酮，具有多种生物作用。然而，尚不清楚大豆蛋白的有益作用是由于染料木黄酮还是其他成分。此外，豆制品血管保护作用的细胞或分子机制仍不清楚。我们最近发现金雀异黄素直接作用于血管内皮细胞（EC），导致cAMP积累、一氧化氮合酶（eNOS）激活和表达。此外，我们证明金雀异黄素可诱导大鼠全身血压（BP）快速降低。因此，本申请的目的是确定金雀异黄酮如何触发 cAMP 信号通路，从而影响 cAMP 相关的血管功能。将解决以下四个问题：1）。金雀异黄素如何提高 EC 中的 cAMP、激活 eNOS，从而刺激一氧化氮？ 2) 金雀异黄素是否通过激活 cAMP 信号来降低动物血压？ 3）。金雀异黄素升高的 cAMP 是否也会改变 eNOS 基因表达？ 4).金雀异黄素是否通过 cAMP 信号传导保护内皮屏障功能？我们将采用细胞、分子和生化分析来确定金雀异黄素是否通过激活 EC 中 Gs 介导的腺苷酸环化酶来升高 cAMP。金雀异黄素调节的 eNOS 基因表达将通过核糖核酸酶保护测定和实时 PCR 进行评估。金雀异黄素对内皮屏障功能的影响（通过细胞骨架重组和屏障通透性确定）将使用免疫细胞化学、共聚焦显微镜和大分子渗漏方法进行评估。将使用药理学和分子干预方法来确定特定途径参与金雀异黄素作用的介导。这些研究的结果将建立金雀异黄酮在脉管系统中的作用机制，从而为进一步研究确定金雀异黄酮对血管健康和高血压等疾病的影响奠定基础。由于异黄酮被广泛用作膳食补充剂，大豆异黄酮对脉管系统的健康益处的证明预计将对公众健康产生重大影响。