Molecular Mechanisms of Genistein in the Prevention of Inflammatory Cytokine (TNF

金雀异黄素预防炎症细胞因子（TNF）的分子机制

• 批准号: 7879814
• 负责人: Zhenquan Jia
• 金额: $ 5.3万
• 依托单位: VIRGINIA COLLEGE OF OSTEOPATHIC MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7879814
• 关键词: Adhesions; Anti-Inflammatory Agents; Anti-inflammatory; Atherosclerosis; Binding; Biological Assay; Blood; Blood Vessels; CD40 Ligand; Cell Adhesion Molecules; Cell Communication; Cell membrane; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Development; Diet; Dietary intake; E-Selectin; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Estrogen Receptors; Functional disorder; GTP-Binding Proteins; Genetic Transcription; Genistein; Goals; Health; Human; Inflammatory; Inflammatory Response; Interleukin-6; Interleukin-8; Intervention; Mediating; Molecular; Molecular Mechanisms of Action; Monocyte Chemoattractant Protein-1; Morbidity - disease rate; Mus; NF-kappa B; Oxidative Stress; Pathogenesis; Pathway interactions; Physiological; Play; Prevention; Production; Protein Tyrosine Kinase; Proteins; Recruitment Activity; Reporter Genes; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Transduction; Soybeans; System; Transcriptional Regulation; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Umbilical vein; Vascular Cell Adhesion Molecule-1; Vascular Diseases; Western Blotting; Work; base; chemokine; cytokine; design; dietary supplements; fundamental research; improved; mRNA Expression; monocyte; mortality; novel; prevent; protective effect; public health relevance; transcription factor; vascular inflammation

DESCRIPTION (provided by applicant): The long-range goal of this research is to identify bioactive components and develop novel nutraceuticals that may improve vascular function and therefore decrease the morbidity and mortality related to vascular dysfunction and related complications. Vascular inflammation and its subsequent endothelial dysfunction play a fundamental role in the initiation and progression of atherosclerosis. It is believed that tumor necrosis factor (TNF)-alpha is critically involved in the pathogenesis of atherosclerosis. Studies show that genistein may have vascular protective effects. However, the molecular mechanism of genistein action is not well understood. The main objective of this study is to evaluate the cellular mechanism of action of genistein in its protective effect on (TNF)-alpha-induced vascular dysfunction. We have collected preliminary data, which suggest a potential anti-inflammatory action of genistein against vascular dysfunction: 1) genistein, at physiological concentrations, significantly inhibited TNF-alpha-induced adhesion of monocytes to human umbilical vein endothelial cells (HUVECs), 2) genistein significantly suppressed TNF-alpha-induced production of monocyte chemoattractant protein -1 (MCP-1) and interleukin-8 (IL-8), two chemokines that are the key factors in the firm adhesion of monocytes to activated endothelial cells, (3) TNF-alpha significantly increased NF-KB binding activity indicating that activation of the transcription factor NF-kB might be critical for the TNF-alpha-induced inflammatory response. We have recently demonstrated that genistein directly activates the cAMP/protein kinase A (PKA) signaling in endothelial cells, which is not related to any known genistein action such as inhibition of tyrosine kinase or binding to estrogen receptors. Our working hypothesis is that genistein, at physiological concentrations, suppresses TNF-alpha-induced recruit of monocytes to endothelial cells via activation of the cAMP/PKA signaling (involving modulation of plasma membrane associated G-proteins) that subsequently inhibits NF-kB activity and its regulated chemokine and adhesion molecular expression. Accordingly, the specific aims of this application are: 1) to determine whether genistein inhibits TNF-alpha-induced expression of adhesion molecules, markers of vascular inflammation and NF-kB in HUVECs, 2) to determine whether the effects of genistein on TNF- alpha-induced monocyte adhesion to endothelial cells, expression of chemokine, adhesion molecules as well as NF-KB activation is mediated through the cAMP/PKA pathway, and whether stimulation of cAMP/PKA cascade by genistein is mediated through G proteins, 3) to investigate whether dietary intake of genistein protects against TNF-alpha-induced vascular inflammation in mice. Fulfillment of the above aims will establish the cellular and molecular mechanisms of action of genistein in its protective effect against vascular dysfunction. PUBLIC HEALTH RELEVANCE: Atherosclerotic vascular disease is a major cause of morbidity and mortality in the industrial world. Our preliminary data indicate genistein, a soybean compound, is a promising agent to protect against vascular dysfunction caused by the proinflammatory cytokine tumor necrosis factor (TNF)-alpha. The elucidation of the molecular mechanism(s) of action of genistein will provide clinically related strategies in protecting against atherosclerotic vascular disease.

描述（由申请人提供）：本研究的长期目标是鉴定生物活性成分并开发可能改善血管功能的新型营养药品，从而降低与血管功能障碍和相关并发症相关的发病率和死亡率。血管炎症及其随后的内皮功能障碍在动脉粥样硬化的发生和发展中起着重要作用。人们认为肿瘤坏死因子(TNF)- α在动脉粥样硬化的发病过程中起着至关重要的作用。研究表明染料木素可能具有血管保护作用。然而，染料木素作用的分子机制尚不清楚。本研究的主要目的是评价染料木素对(TNF)- α诱导的血管功能障碍的保护作用的细胞机制。我们收集了初步数据，表明染料木素对血管功能障碍具有潜在的抗炎作用：1)在生理浓度下，染料木素显著抑制tnf - α诱导的单核细胞与人脐静脉内皮细胞（HUVECs）的粘附；2)染料木素显著抑制tnf - α诱导的单核细胞趋化蛋白-1 （MCP-1）和白细胞介素-8 （IL-8）的产生，这两种趋化因子是单核细胞与活化的内皮细胞紧密粘附的关键因素。(3) tnf - α显著增加NF-KB结合活性，表明转录因子NF-KB的激活可能对tnf - α诱导的炎症反应至关重要。我们最近证明染料木素直接激活内皮细胞中的cAMP/蛋白激酶A （PKA）信号，这与任何已知的染料木素作用如抑制酪氨酸激酶或结合雌激素受体无关。我们的工作假设是，在生理浓度下，染料木素通过激活cAMP/PKA信号（包括调节质膜相关g蛋白）抑制tnf - α诱导的单核细胞向内皮细胞的募集，进而抑制NF-kB活性及其调节的趋化因子和粘附分子表达。因此，这一应用的具体目的是：1)确定染料木素是否抑制TNF- α诱导的内皮细胞粘附分子、血管炎症标志物和NF-kB的表达；2)确定染料木素对TNF- α诱导的内皮细胞单核细胞粘附、趋化因子表达、粘附分子以及NF-kB活化的影响是否通过cAMP/PKA通路介导，染料木素刺激cAMP/PKA级联是否通过G蛋白介导；3)探讨膳食摄入染料木素是否对小鼠tnf - α诱导的血管炎症有保护作用。上述目的的实现将为染料木素抗血管功能障碍的细胞和分子机制的建立奠定基础。