Inhibitor ameliorates oxidative and proteolytic strees

抑制剂可改善氧化和蛋白水解应激

• 批准号: 7340811
• 负责人: Suresh C. Tyagi
• 金额: $ 9.23万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-05 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7340811
• 关键词: 3-nitrotyrosine; Abbreviations; Acetylcholine; Acids; Antibodies; Antigens; Arginine; Arteriovenous fistula; Biochemical; Bradykinin; CD31 Antigens; Cardiac; Cells; Chronic; Collagen; Complex; Congestive; Congestive Heart Failure; Diastolic blood pressure; Disulfides; Endothelial Cells; Endothelium; Extracellular Matrix; Feedback; Functional disorder; Goals; Heart; Heart failure; Hydroxyeicosatetraenoic Acids; Hypertrophy; In Situ; Ischemia; Left; Left ventricular structure; Matrix Metalloproteinases; Measures; Metalloproteases; Methods; Molecular; Mus; Muscle; Myocardial; NADH; Nicotinamide adenine dinucleotide; Nitric Oxide; Nitric Oxide Donors; Nitroprusside; Nuclear; Oxidases; Oxidation-Reduction; Oxidative Stress; Oxidative Stress Induction; Oxides; Peroxisome Proliferator-Activated Receptors; Peroxonitrite; Physiological; Physiological reperfusion; Plasma; Poly(ADP-ribose) Polymerases; Principal Investigator; Prostaglandins; Protein Kinase C; Proteins; Purpose; Reactive Oxygen Species; Reperfusion Therapy; Reporting; Repression; Role; Stress; Sulfhydryl Compounds; Superoxide Dismutase; Techniques; Testing; Tissue Inhibitor of Metalloproteinases; Tissues; Tyrosine; Ventricular; Western Blotting; arginine methyl ester; catalase; density; human NOS3 protein; improved; inhibitor/antagonist; prevent; progesterone 11-hemisuccinate-(2-iodohistamine); programs; research study; response; tetrahydrobiopterin; tissue inhibitor of metalloproteinase 4

Endocardial endothelial (EE) dysfunction and accumulation of oxidized-matrix between endothelial and muscle are the hallmarks of congestive heart failure (CHF). The overall objective of this project is to understand the mechanism of oxidized-matrix accumulation and EE dysfunction in CHF. Previous studies have suggested an association between induction of oxidative stress, decrease in endothelial cell density, activation of matrix metalloproteinase (MMP), collagenolysis, and repression of cardiac tissue inhibitor of metalloproteinase (CIMP) in CHF. The novelty of this proposal is that among all known tissue inhibitors of metalloproteinase (TIMP), the TIMP-4 (i.e. CIMP) is highly expressed in the heart. The purpose of this proposal is to test a central hypothesis that oxidized-matrix accumulation and EE dysfunction are due to increased levels of MMP activity, and collagenolysis. These levels are associated with decreased levels of EE nitric oxide, and CIMP in response to increased levels of reactive oxygen species (ROS) during protracted cycles of ischemia/reperfusion in volume overload. The increased levels of CIMP protects EE against oxidative and proteolytic stresses. We will test the central hypothesis by the following three specific aims: L To determine whether CIMP decreases oxidative stress by increasing EE nitric oxide concentration. Plasma and left ventricle levels of nitric oxide, ROS, and nitrotyrosine will be measured in chronic volume overload arteriovenous fistula mice treated with and without CIMP. 2_ To determine whether CIMP protein transfer inhibits collagenolysis. MMP activity will be measured by in situ zymography. The levels of CIMP activity will be measured by reverse zymography. Total collagen and its degradation fragments will be measured by Western-blot analysis using anti-collagen antibody. 3-- To determine whether CIMP ameliorates endocardial endothelial dysfunction. Contractile responses to acetylcholine, bradykinin, and nitroprusside in cardiac rings will be measured in a tissue myobath. These studies will enable us to determine whether CIMP improves the hearts response to nitric oxide donors. Identification of major players involved in the control of oxidative and proteolytic stresses will help to develop strategies to prevent CHF.

心内膜内皮 (EE) 功能障碍以及内皮和肌肉之间氧化基质的积累 是充血性心力衰竭（CHF）的标志。该项目的总体目标是了解 CHF 中氧化基质积累和 EE 功能障碍的机制。先前的研究表明 氧化应激诱导、内皮细胞密度降低、基质活化之间的关联 金属蛋白酶 (MMP)、胶原溶解和金属蛋白酶的心脏组织抑制剂的抑制 (CIMP) 换算为瑞士法郎。该提案的新颖之处在于，在所有已知的金属蛋白酶组织抑制剂中 (TIMP)，TIMP-4（即 CIMP）在心脏中高表达。该提案的目的是测试 中心假设是氧化基质积累和 EE 功能障碍是由于 MMP 水平升高所致 活性和胶原蛋白溶解。这些水平与 EE 一氧化氮和 CIMP 水平降低有关 响应延长周期期间活性氧 (ROS) 水平的增加 容量超负荷时的缺血/再灌注。 CIMP 水平的增加可保护 EE 免受氧化和 蛋白水解应激。我们将通过以下三个具体目标来检验中心假设： L To 确定 CIMP 是否通过增加 EE 一氧化氮浓度来降低氧化应激。等离子体 慢性容量超负荷时将测量左心室一氧化氮、活性氧和硝基酪氨酸的水平 使用和不使用 CIMP 治疗的动静脉瘘小鼠。 2_判断CIMP蛋白是否转移 抑制胶原蛋白溶解。 MMP 活性将通过原位酶谱法测量。 CIMP 活动水平 将通过反向酶谱法进行测量。总胶原蛋白及其降解片段将通过以下方式测量 使用抗胶原抗体进行蛋白质印迹分析。 3--确定CIMP是否改善 心内膜内皮功能障碍。对乙酰胆碱、缓激肽和硝普钠的收缩反应 心环将在组织肌浴中测量。这些研究将使我们能够确定 CIMP 是否 改善心脏对一氧化氮供体的反应。识别参与控制的主要参与者 氧化应激和蛋白水解应激将有助于制定预防 CHF 的策略。