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

基本信息

批准号：
7816316
负责人：
DONGMIN LIU
金额：
$ 21.31万
依托单位：
VIRGINIA POLYTECHNIC INST AND ST UNIV
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7816316
关键词：
Accounting Adenovirus Protein Adenylate Cyclase Adhesions Affect American Animals Anti-Inflammatory Agents Anti-inflammatory Atherosclerosis Attenuated Biological Assay Blood Glucose Blood Vessels Botanicals Cardiovascular Diseases Catalytic Domain Cause of Death Cell Adhesion Cell Adhesion Molecules Cell Communication Cell Culture Techniques Cessation of life Chemicals Chronic Chronic Disease Cyclic AMP Cyclic AMP-Dependent Protein Kinases DNA Binding Data Development Diabetes Mellitus Diabetic Angiopathies Diabetic mouse Dietary Supplementation Dietary intake E-Selectin Endothelial Cells Endothelium Environment Enzyme-Linked Immunosorbent Assay Equipment Estrogen Receptors Experimental Designs Fabaceae Functional disorder Funding Gene Expression Genes Genistein Glucose Goals Human Human Resources Hyperglycemia Image Immunoblot Analysis In Vitro Incubated Incubators Inflammation Intercellular adhesion molecule 1 Interleukin-8 Intervention Knockout Mice Lead Leukocytes Ligand Binding Ligands Measures Mediating Methods Microscope Molecular Monocyte Chemoattractant Protein-1 Morbidity - disease rate Nuclear Nutritional Obesity Parents Pathogenesis Pathway interactions Patients Peroxisome Proliferator-Activated Receptors Physiological Plasma Play Postdoctoral Fellow Prevention Production Protein Kinase A Inhibitor Protein Tyrosine Kinase Proteins Protocols documentation Recovery Recruitment Activity Reporter Genes Research Research Contracts Research Design Research Personnel Role Schedule Signal Pathway Signal Transduction Small Interfering RNA Streptozocin Supplementation System Testing Therapeutic Therapeutic Agents Time Up-Regulation Vascular Endothelial Cell Western Blotting adenylate base chemokine cost diabetic diabetic patient exposed human population flexibility in vivo instrument intraperitoneal knockout animal mRNA Expression methylxanthine monocyte mortality new technology novel parent grant parent project prevent protein expression public health relevance research study tool vascular inflammation

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this research is to decrease the morbidity and mortality related to diabetic vascular complications by developing 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 monocyte-EC interaction, suggesting an anti-inflammatory action of genistein in vasculature of diabetics. 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. Our recent studies found that genistein is an activator of the cAMP signaling system in ECs. In addition, our data demonstrated that genistein can stimulate PPAR3 activity in HAECs. Interestingly, data from our animal study showed that dietary intake of genistein increased plasma cAMP but reduced MCP-1 levels in diabetic mice. Our specific aim of this application is to determine the molecular mechanism by which genistein suppresses hyperglycemia-induced vascular inflammation. We hypothesized that genistein inhibits hyperglycemia-induced EC inflammation by activation of cAMP/PKA signaling and PPAR3 in ECs. Aim 1 will use HAECs to investigate: 1) whether genistein inhibits high glucose-induced expression of chemokines and adhesion molecules; 2) whether genistein inhibits hyperglycemia-induced NF-kB activity; 3) whether the anti-inflammatory action of genistein is mediated via the cAMP/PKA and PPAR3 pathways; and 4) whether genistein also inhibits hyperglycemia-induced EC-monocyte interaction under flow. Aim 2 will use diabetic mice to determine: 1) whether genistein activates cAMP/PKA and PPAR3 in vivo; 2) whether genistein inhibits chemokine and adhesion molecule production from blood vessels; and 3) whether genistein suppresses monocyte recruitment to ECs and whether this effect is attenuated by PPAR3 deficiency. Genistein will be administered to diabetic mice through dietary supplementation approach. The protein and mRNA expression will be assessed by Western blot, ELISA or real-time PCR. Cyclic AMP levels and PKA activity will be determined using assay kits. NF-kB and PPAR3 activity will be measured with ELISA and reporter gene assays. The role of PKA and PPAR3 in the anti-inflammatory action of genistein will be determined by using chemical and molecular tools as well as gene knockout mice. 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 genisein 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 23 million Americans.

描述（由申请人提供）：这项研究的长期目标是通过开发生物活性治疗剂来降低与糖尿病血管并发症相关的发病率和死亡率。高血糖引起的血管炎症，导致对血管内皮细胞（ECS）的单核细胞粘附，是启动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，植物染料蛋白酶抑制了高血糖诱导的人类单核细胞-EC相互作用，这表明染料木黄酮在糖尿病患者的脉管系统中的抗炎作用。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，这些分子的表达受到核因子KB（NF-KB）的严格调节。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蛋白1（MCP-1）和白介素8（IL-8）的产生，两种趋化因子是企业粘附的关键因子单核细胞至激活的EC。我们最近的研究发现，Genastein是EC中cAMP信号系统的激活剂。此外，我们的数据表明，染料木黄酮可以刺激HAEC中的PPAR3活性。有趣的是，我们动物研究的数据表明，染料木黄酮的饮食摄入增加了血浆cAPP，但糖尿病小鼠的MCP-1水平降低。该应用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Genastein通过在EC中激活CAMP/PKA信号传导和PPAR3抑制高血糖诱导的EC炎症。 AIM 1将使用HAEC进行调查：1）染料木黄酮是否抑制了高葡萄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）染料木黄酮的抗炎作用是否是通过CAMP/PKA和PPAR3途径介导的； 4）染料木黄酮是否还抑制流量下高血糖诱导的EC形成细胞相互作用。 AIM 2将使用糖尿病小鼠来确定：1）染料木黄酮在体内是否激活CAMP/PKA和PPAR3； 2）染料木黄酮是否抑制趋化因子和血管产生粘附分子； 3）染料木黄酮是否抑制了对EC的单核细胞募集，以及这种作用是否因PPAR3缺乏症而减弱。染料木黄酮将通过饮食补充方法对糖尿病小鼠施用。蛋白质和mRNA表达将通过Western印迹，ELISA或实时PCR进行评估。循环AMP水平和PKA活性将使用测定试剂盒确定。 NF-KB和PPAR3活性将用ELISA和报告基因测定法测量。 PKA和PPAR3在染料木黄酮的抗炎作用中的作用将通过使用化学和分子工具以及基因基因敲除小鼠来确定。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，植物吉妮丝蛋白是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2300万美国人的慢性疾病。