Mechanism of Vascular Remodeling in Hyperhomocysteinemia

高同型半胱氨酸血症血管重塑机制

• 批准号: 7810738
• 负责人: Suresh C. Tyagi
• 金额: $ 37万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7810738
• 关键词: 3-nitrotyrosine; 9-deoxy-delta-9-prostaglandin D2; Abbreviations; Acetylcholine; Address; Agonist; Aorta; Arachidonic Acids; Attenuated; Basement membrane; Biological Assay; Blood Vessels; Blood flow; Bradykinin; Breeding; Caliber; Collagen; Cystathionine; Disease; Disintegrins; Dose; EMSA; Eicosatrienoic Acid; Elastin; Electrodes; Endothelin-1; Extracellular Matrix; Fibrosis; Fluorescein; Fluoresceins; Functional disorder; Funding; Gelatin; Gelatinase A; Gelatinase B; Genes; Genetic; Goals; Heterozygote; Homo; Homocysteine; Homocystine; Homozygote; Hydroxyeicosatetraenoic Acids; Hyperhomocysteinemia; In Situ; Knock-out; Knockout Mice; Leukotriene B4; Matrix Metalloproteinases; Measures; Mediating; Metalloproteases; Mitochondria; Mus; NADH; NADH oxidase; NF-kappa B; NG-Nitroarginine Methyl Ester; Nicotinamide adenine dinucleotide; Nitric Oxide; Nitroprusside; Nuclear; Nuclear Receptors; Oxidation-Reduction; Oxidative Stress; Peroxisome Proliferator-Activated Receptors; Plasma; Polymerase Chain Reaction; Proteomics; RXR; Reactive Oxygen Species; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; Smooth Muscle; Smooth Muscle Myocytes; Stress; Structure; Superoxide Dismutase; Therapeutic; Thick; Thioredoxin; Time; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinases; Tissues; Vascular Diseases; Vascular remodeling; Western Blotting; arginine methyl ester; arterial remodeling; ciglitazone; ciprofibrate; human AKAP13 protein; human NOS3 protein; in vivo; insight; mouse model; novel; peroxiredoxin; pressure; response; tetrahydrobiopterin; tissue inhibitor of metalloproteinase 4; vascular endothelial dysfunction; vasoactive agent

DESCRIPTION (provided by applicant): Elevated plasma levels of homocysteine (Hcy) known as hyperhomocysteinemia (HHcy) cause vascular disease. The long-term objective of this project is to understand the mechanism of vascular remodeling in HHcy. Studies from the previous funding period showed that there is an inverse relationship between plasma Hcy and levels of peroxisome proliferators activated receptor (PPAR), a nuclear receptor which ameliorates vascular endothelial dysfunction. The central hypothesis of this competitive renewal application is that Hcy decreases thioredoxin, peroxiredoxin, increases NADH oxidase, mtNOS activity, and reactive oxygen species (ROS) in mitochondria in a gene dose-dependent manner. ROS transduces metalloproteinase activation causing thickening (fibrosis) of the basement membrane, rendering ineffective endothelial nitric oxide synthase (eNOS) and promotes endothelial-smooth muscle disconnection/uncoupling by antagonizing PPARy activity. The specific aims to address the central hypothesis are as following: Specific Aim #1: To determine whether by rendering ineffective PPARy Hcy decreases mitochondrial thioredoxin, peroxiredoxin, increases NADH oxidase, mtNOS activity and ROS. The PPARy agonist, ciglitazone will be administered to wild type (WT), cystathione (3 synthase (CBS) knockout heterozygote and homozygote (i.e. CBS +/+, CBS - /+ and CBS -/-), PPARy-/+, iNOS-/-, p47-/-, MMP-9-/- and double knockout mice. Aortic PPARy activity will be measured by EMSA. Levels of mitochondrial ROS will be measured using 2, 7-dichlorofluorescein in continuous assay. Peroxiredoxin, thioredoxin and NADH oxidase will be measured in situ, by Western blot and RT-PCR analyses. Specific Aim #2: To determine whether Hcy increases metalloproteinase activity, decreases the elastin/collagen ratio, and causes fibrosis by antagonizing PPARy. Metalloproteinase, TIMP, elastin and collagen will be measured in situ, by novel 2-D zymography, reverse-zymography, real-time RT- PCR and Western blot analysis. Specific Aim #3: To determine whether Hcy attenuates vascular function by increasing iNOS and rendering eNOS ineffective by antagonizing PPARy. Aortic contractile response will be measured. The novelty of this proposal is that it elucidates the mitochondrial mechanism of oxidative stress and vascular remodeling by Hcy. This study will provide new insights into the mechanism of arterial remodeling and will have therapeutic ramifications for vessel wall disease.

描述（由申请人提供）：同型半胱氨酸（HCY）的血浆水平升高，称为高型半胱氨酸血症（HHCY）引起血管疾病。该项目的长期目标是了解HHCY中血管重塑的机制。前一个资金期的研究表明，血浆HCY与过氧化物酶体增生剂的水平之间存在反比关系，这是一种核受体，可改善血管内皮功能障碍。这种竞争性更新应用的中心假设是，HCY以基因剂量依赖性方式在线粒体中降低了硫氧还蛋白，过氧蛋白，增加NADH氧化酶，mTNOS活性和活性氧（ROS）。 ROS传递金属蛋白酶活化，导致基底膜的增厚（纤维化），使无效的内皮一氧化氮合酶（ENOS）呈现，并促进内皮平滑的肌肉断开/通过拮抗Ppary活性来偶联。解决中心假设的具体目的如下：特定目的＃1：通过呈现无效的PPARY HCY是否会降低线粒体硫氧还蛋白，过氧蛋白，增加NADH氧化酶，mTNOS活性和ROS。 The PPARy agonist, ciglitazone will be administered to wild type (WT), cystathione (3 synthase (CBS) knockout heterozygote and homozygote (i.e. CBS +/+, CBS - /+ and CBS -/-), PPARy-/+, iNOS-/-, p47-/-, MMP-9-/- and double knockout mice. Aortic PPARy通过在连续测定中，将通过2、7-二氯氟乙烯素来测量线粒体ROS的水平。通过抗抗蛋白酶，抗蛋白酶，TIMP，弹性蛋白和胶原蛋白将通过新的2-D Zymography，实时的RT-PCR和Western Blot分析来测量该建议阐明了HCY的氧化应激和血管重塑的线粒体机制。

项目成果

Congenic expression of tissue inhibitor of metalloproteinase in Dahl-salt sensitive hypertensive rats is associated with reduced LV hypertrophy.

• DOI: 10.1080/13813450802535978
• 发表时间: 2008-12
• 期刊: Archives of physiology and biochemistry
• 影响因子: 3
• 作者: Rodriguez WE;Tyagi N;Deng AY;Adeagbo A;Joshua IG;Tyagi SC
• 通讯作者: Tyagi SC

Folic acid improves inner ear vascularization in hyperhomocysteinemic mice.

• DOI: 10.1016/j.heares.2011.12.006
• 发表时间: 2012-02
• 期刊: HEARING RESEARCH
• 影响因子: 2.8
• 作者: Kundu, Soumi;Munjal, Charu;Tyagi, Neetu;Sen, Utpal;Tyagi, Aaron C.;Tyagi, Suresh C.
• 通讯作者: Tyagi, Suresh C.

Folic acid improves acetylcholine-induced vasoconstriction of coronary vessels isolated from hyperhomocysteinemic mice: an implication to coronary vasospasm.

• DOI: 10.1002/jcp.22621
• 发表时间: 2011-10
• 期刊: JOURNAL OF CELLULAR PHYSIOLOGY
• 影响因子: 5.6
• 作者: Qipshidze, Natia;Metreveli, Naira;Lominadze, David;Tyagi, Suresh C.
• 通讯作者: Tyagi, Suresh C.

Fibrinogen induces alterations of endothelial cell tight junction proteins.

• DOI: 10.1002/jcp.21845
• 发表时间: 2009-10
• 期刊: JOURNAL OF CELLULAR PHYSIOLOGY
• 影响因子: 5.6
• 作者: Patibandla, Phani K.;Tyagi, Neetu;Dean, William L.;Tyagi, Suresh C.;Roberts, Andrew M.;Lominadze, David
• 通讯作者: Lominadze, David

Differential expression of gamma-aminobutyric acid receptor A (GABA(A)) and effects of homocysteine.

γ-氨基丁酸受体 A (GABA(A)) 的差异表达和同型半胱氨酸的影响。

• DOI: 10.1515/cclm.2007.342
• 发表时间: 2007
• 期刊: Clinical chemistry and laboratory medicine
• 影响因子: 6.8
• 作者: Tyagi,Neetu;Lominadze,David;Gillespie,William;Moshal,KarniS;Sen,Utpal;Rosenberger,DorotheaS;Steed,Mesia;Tyagi,SureshC
• 通讯作者: Tyagi,SureshC