Genistein for the Prevention and Treatment of Diabetic Vascular Inflammation

金雀异黄酮预防和治疗糖尿病血管炎症

• 批准号: 7686348
• 负责人: DONGMIN LIU
• 金额: $ 23.78万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7686348
• 关键词: Accounting; Adenovirus Protein; Adenylate Cyclase; Adhesions; Affect; American; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Atherosclerosis; Biological Assay; Blood Glucose; Blood Vessels; Botanicals; Budgets; Cardiovascular Diseases; Catalytic Domain; Cause of Death; Cell Adhesion; Cell Adhesion Molecules; Cell Communication; Cessation of life; Chronic; Chronic Disease; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Development; Diabetes Mellitus; Diabetic Angiopathies; Diabetic mouse; Dietary intake; E-Selectin; Endothelial Cells; Endothelium; Enzyme-Linked Immunosorbent Assay; Estrogen Receptors; Fabaceae; Functional disorder; Gene Expression; Genes; Genistein; Glucose; Goals; Grant; Human; Hyperglycemia; Intercellular adhesion molecule 1; Interleukin-8; Intervention; Laboratory Finding; Lead; Leukocytes; Mediating; Molecular; Monocyte Chemoattractant Protein-1; Morbidity - disease rate; National Center for Complementary and Alternative Medicine; Nuclear; Nutraceutical; Nutritional; Obesity; Pathogenesis; Pathway interactions; Patients; Peroxisome Proliferator-Activated Receptors; Physiological; Plasma; Play; Prevention; Production; Property; Protein Kinase A Inhibitor; Protein Tyrosine Kinase; Proteins; Protocols documentation; Recruitment Activity; Reporter Genes; Research; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; Streptozocin; Supplementation; System; Testing; Therapeutic; Therapeutic Agents; Time; Vascular Endothelial Cell; Western Blotting; adenylate; base; chemokine; cost; diabetic; diabetic patient; exposed human population; in vivo; innovation; intraperitoneal; mRNA Expression; methylxanthine; monocyte; mortality; mouse model; novel; prevent; protective effect; public health relevance; systems research; vascular inflammation

DESCRIPTION (provided by applicant): The long-term goal of this research is to decrease the morbidity and mortality related to diabetic vascular dysfunction and related complications, by developing novel, bioactive therapeutic agents. Hyperglycemia- induced vascular inflammation, leading to monocyte adhesion to vascular endothelial cells (ECs), is the key factor that initiates the pathogenesis of atherosclerosis in diabetic patients. Our studies found that botanical genistein inhibits hyperglycemia-induced human leukocyte-EC interaction, suggesting a potential anti- inflammatory action of genistein in diabetic vascular complications. Leukocyte adhesion to endothelium is mediated through both chemokines and adhesion molecules on ECs, and the expression of these molecules is critically up-regulated by nuclear factor kB (NF-kB). Further analysis showed that genistein suppressed high glucose-induced production of monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) in human aortic ECs (HAECs), two chemokines that are the key factors in the firm adhesion of monocytes to activated ECs. Recent studies in this laboratory found that genistein is an activator of the cAMP signaling system in ECs. Interestingly, our studies showed that dietary intake of genistein increased plasma cAMP but reduced MCP-1 levels in diabetic mice. Our specific aim of this proposal is to determine the molecular mechanism by which genistein suppresses hyperglycemia-induced vascular inflammation. We hypothesized that genistein, by activation of the cAMP/PKA signaling, inhibits high glucose-induced monocyte adhesion to ECs through suppression of NF-kB activity and subsequent inhibition of chemokine and adhesion molecule expression. Specific aim 1 will use HAECs to investigate: 1) whether genistein inhibits hyperglycemia-induced expression of chemokines and adhesion molecules; 2) whether genistein inhibits hyperglycemia-induced NF-kB activity; and 3) whether the anti-inflammatory properties of genistein are mediated via the cAMP/PKA pathway. Specific aim 2 will use diabetic mouse models to determine: 1) whether genistein induces cAMP/PKA signaling in vivo; 2) whether genistein inhibits chemokine and adhesion molecule production from blood vessels; and 3) whether genistein suppresses monocyte recruitment to vascular ECs. Genistein will be administered through a nutritional supplementation approach to both chemically induced and obese diabetic mice. The protein and mRNA expression will be assessed by Western blot, ELISA or real-time PCR. The adenylate cylase activity, cAMP and PKA activation in ECs will be determined using assay kits. NF-kB activity will be determined with ELISA and reporter gene assay. The role of the cAMP/PKA cascade in the anti-inflammatory action of genistein will be determined by using both pharmacological and molecular intervention approaches. The mechanistic understanding of how genistein protects against diabetes-caused vascular inflammation will facilitate the development of efficacious CAM strategies to protect diabetic patients from vascular complications. PUBLIC HEALTH RELEVANCE: Cardiovascular disease such as atherosclerosis is the main cause of deaths in patients with diabetes mellitus. It is increasingly recognized that vascular inflammation mediated by sustained elevation of blood glucose levels in diabetic patients plays a pivotal role in the pathogenesis of atherosclerosis. Our studies indicate that botanical genistein is a promising agent to protect against chronic high glucose- caused vascular inflammation. The results of this research will potentially lead us to develop novel, natural, and cost-effective agents to prevent and treat atherosclerosis caused by diabetes, a chronic disease that affects over 20 million Americans.

描述（由申请人提供）：这项研究的长期目标是通过开发新型的生物活性治疗剂来降低与糖尿病血管功能障碍和相关并发症相关的发病率和死亡率。高血糖诱导的血管炎症，导致对血管内皮细胞（ECS）的单核细胞粘附，是引发糖尿病患者动脉粥样硬化发病机理的关键因素。我们的研究发现，植物染料木黄酮抑制了高血糖诱导的人白细胞-EC相互作用，这表明染料木黄酮在糖尿病血管并发症中的潜在抗炎作用。白细胞对内皮的粘附是通过趋化因子和EC上的粘附分子介导的，这些分子的表达被核因子KB（NF-KB）严重上调。进一步的分析表明，染料木黄酮抑制了人主动脉ECS（HAECS）中高葡萄糖诱导的单核细胞趋化蛋白1（MCP-1）和白介素-8（IL-8）的产生，这是两种趋化因子，这是单核细胞粘附于单核细胞粘附到激活的ECS的关键因素。该实验室的最新研究发现，染料木黄酮是EC中cAMP信号系统的激活因子。有趣的是，我们的研究表明，饮食中染料木黄酮的摄入增加了血浆营地，但糖尿病小鼠的MCP-1水平降低。该提议的具体目的是确定染料木黄酮抑制高血糖诱导的血管炎症的分子机制。我们假设通过激活CAMP/PKA信号传导，染料木黄酮通过抑制NF-KB活性并随后抑制趋化因子和粘附分子表达来抑制高葡萄糖诱导的单核细胞粘附于EC。具体目标1将使用HAEC进行调查：1）染料木黄酮是否抑制高血糖诱导的趋化因子和粘附分子的表达； 2）染料木黄酮是否抑制高血糖诱导的NF-KB活性； 3）染料木黄酮的抗炎特性是否是通过CAMP/PKA途径介导的。特定的目标2将使用糖尿病小鼠模型来确定：1）染料木黄酮是否在体内诱导CAMP/PKA信号传导； 2）染料木黄酮是否抑制趋化因子和血管产生粘附分子； 3）染料木黄酮是否会抑制单核细胞募集到血管EC中。染料木黄酮将通过对化学诱导和肥胖的糖尿病小鼠的营养补充方法来施用。蛋白质和mRNA表达将通过Western印迹，ELISA或实时PCR进行评估。 EC中的腺苷酸元酶活性，CAMP和PKA激活将使用测定试剂盒确定。 NF-KB活性将用ELISA和报告基因测定法确定。 CAMP/PKA级联在染料木黄酮的抗炎作用中的作用将通过使用药理和分子干预方法来确定。对染料木黄酮如何保护糖尿病引起的血管炎症的机械理解将有助于开发有效的相机策略，以保护糖尿病患者免受血管并发症的侵害。 公共卫生相关性：诸如动脉粥样硬化之类的心血管疾病是糖尿病患者死亡的主要原因。人们越来越认识到，糖尿病患者血糖水平持续升高介导的血管炎症在动脉粥样硬化的发病机理中起关键作用。我们的研究表明，植物染料木黄酮是一种预防慢性高葡萄糖引起的血管炎症的有前途的药物。这项研究的结果可能会导致我们开发新颖，自然和具有成本效益的药物，以预防和治疗由糖尿病引起的动脉粥样硬化，糖尿病是一种影响超过2000万美国人的慢性疾病。