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.

心内膜内皮细胞功能障碍与内皮与肌肉间氧化基质堆积 是充血性心力衰竭(CHF)的特征。本项目的总体目标是了解 心力衰竭时氧化基质堆积和EE功能障碍的机制。之前的研究表明， 氧化应激诱导、内皮细胞密度降低、基质激活之间的关系 金属蛋白酶、胶原酶溶解和心脏组织金属蛋白酶抑制物抑制 (CIMP)(CHF)。这项提议的新奇之处在于，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。这项提议的目的是测试一个 氧化基质堆积和EE功能障碍是由于基质金属蛋白酶水平升高的中心假说 活性和胶原蛋白分解。这些水平与EE、NO和CIMP水平的降低有关 在长时间的循环中，由于活性氧物种(ROS)水平的增加， 容量超负荷的缺血/再灌流。CIMP水平升高可保护EE免受氧化和 蛋白水解性压力。我们将通过以下三个具体目标来检验中心假设：L 确定CIMP是否通过增加EE一氧化氮浓度来降低氧化应激。电浆 慢性容量超负荷患者的左心室一氧化氮、ROS和硝基酪氨酸水平将被测量 CIMP治疗和不治疗动静脉瘘小鼠。2_确定CIMP蛋白转移 抑制胶原蛋白的分解。用原位酶谱法检测基质金属蛋白酶活性。CIMP活性水平 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 用抗胶原抗体进行蛋白印迹分析。3--确定CIMP是否有所改善 心内膜内皮细胞功能障碍。血管对乙酰胆碱、缓激肽和硝普钠的收缩反应 心脏环将以组织肌层为单位进行测量。这些研究将使我们能够确定CIMP 改善心脏对一氧化氮供体的反应。确定参与控制的主要参与者 氧化和蛋白分解应激将有助于制定预防心力衰竭的策略。