Exercise, Diabetes, & Coronary Smooth Muscle Ca2+

运动，糖尿病，

基本信息

批准号：
8220817
负责人：
Michael Sturek
金额：
$ 42.69万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-04-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8220817
关键词：
Acute Adenosine Adenosine A1 Receptor Aerobic Exercise Agonist Aldosterone Angiotensin II Angiotensin Receptor Angiotensins Animal Model Antisense Oligonucleotides Aorta Arterial Fatty Streak Arteries Atherosclerosis Attenuated Biomedical Engineering Cells Characteristics Clinical Research Collagen Complex Coronary Coronary Arteriosclerosis Coronary Restenosis Coronary Stenosis Coronary artery Coronary heart disease Cytotoxin Deposition Development Devices Diabetes Mellitus Diffuse Dyslipidemias Exercise Experimental Designs Exposure to Family suidae Genes Glucose Intolerance Goals Grant Growth Health Histopathology Human Hyperlipidemia Hypertension Immunoblotting Immunohistochemistry In Vitro Inflammation Injury Insulin Resistance Kidney Lesion Lipids Losartan MAPK8 gene Measures Messenger RNA Metabolic syndrome Metals Mineralocorticoids Mitogens Modeling Molecular Non-Insulin-Dependent Diabetes Mellitus Nucleotides Obesity Organ Culture Techniques P2Y2 receptor Paclitaxel Patients Pharmaceutical Preparations Pharmacotherapy Phenotype Phosphotransferases Platelet aggregation Positioning Attribute Prediabetes syndrome Prevention Procedures Protein Isoforms Proteins Proto-Oncogene Proteins c-akt Purinergic P1 Receptors Receptor Signaling Recovery Regulation Renin Renin-Angiotensin-Aldosterone System Reporting Research Reverse Transcriptase Polymerase Chain Reaction Risk Sarcoplasmic Reticulum Signal Pathway Signal Transduction Sirolimus Small Interfering RNA Smooth Muscle Smooth Muscle Myocytes Spironolactone Stenosis Stents Testing Therapeutic Effect Thrombosis Time Training Translations Ultrasonography Vascular Diseases attenuation clinically relevant clinically significant immunocytochemistry improved in vivo injured migration non-diabetic novel novel therapeutic intervention nucleotide receptor optical imaging overexpression oxidant stress prevent public health relevance receptor receptor expression response restenosis scaffold selective expression

项目摘要

DESCRIPTION (provided by applicant): The long-term goal is to determine the coronary smooth muscle (CSM) adenosine A1 and P2Y2 receptor signaling mechanisms by which antagonism of the renin-angiotensin-aldosterone system (RAAS) and exercise training decrease coronary artery disease (CAD) and restenosis in metabolic syndrome (MetS). RAAS regulation of CSM A1 and P2Y2 receptor signaling that modulates kinases and downstream inflammation is unknown. Although drug-eluting stents decrease restenosis, further study is needed because of restenosis complications and progression of CAD adjacent to the stent, i.e. peri-stent CAD. Major findings in our previous grant provide outstanding rationale for this project. Ossabaw swine have clinically significant atherosclerosis (>50% stenosis) and we have for the first time stented natural lesions, not just balloon-injured healthy arteries. We cloned all 4 adenosine receptor isoforms (A1, A2A, A2B, A3) from pig and found A1 receptors (A1R) selectively expressed in CSM and increased in atherosclerotic and stented coronary. Direct activation of A1R induces CSM proliferation, indicating novel regulation. We cloned 3 porcine P2 nucleotide receptor isoforms and found the P2Y2 subtype is only expressed in diseased CSM and is selectively up-regulated in stented coronary. Overall hypothesis: in MetS RAAS (mainly aldosterone) increases coronary A1R and P2Y2R signaling, pivotal signals for CSM growth. The integrative experimental design compares lean vs. MetS Ossabaw RAAS antagonism. Atherosclerotic lesions are stented followed by recovery with or without exercise. Results are compared to simpler in vitro organ culture. Specific Aims are to test 6 specific hypotheses in MetS vs. lean Ossabaw: 1) Atherosclerosis, peri-stent CAD, and in-stent stenosis are increased in MetS and attenuated by RAAS antagonism and exercise. Intravascular ultrasound assesses CAD in vivo and optical imaging and histopathology assess CAD in vitro. 2) MetS increases molecular expression of the A1R and P2Y2R. Protein and mRNA will be determined in 5 different coronary segments: healthy, atherosclerotic, peri-stent, in-stent neointima, and in-stent media. 3) MetS increases functional expression of the A1R and P2Y2R by ERK, JNK, and AKT activation. Immunoblots and immunocytochemistry for phospho ERK, JNK, and AKT; store-operated Ca influx; and inflammation and oxidant stress quantify functional activation. 4) Angiotensin II and aldosterone directly, independently, and synergistically increase molecular and functional expression of A1R and P2Y2R, effects that are potentiated by dyslipidemia. In vivo treatments include several pig groups to be compared with in vitro organ culture. 5) Exercise will reverse the increased molecular and functional expression of A1R and P2Y2R in MetS. This Aim assesses more established CAD and stenting. 6) Blockade of A1R and P2Y2R attenuates in-stent stenosis in over-expansion injury and native atherosclerosis. Lean and MetS pigs will undergo in vivo coronary stenting with drug- and gene-eluting stents to selectively block A1R and P2Y2R. PUBLIC HEALTH RELEVANCE: Our studies involve highly clinically relevant coronary artery disease in the Ossabaw miniature pig, which superbly mimics complex metabolic syndrome ("pre-diabetes") and type 2 diabetes in humans. Thus, our findings will catalyze the translation of novel therapeutic interventions to humans, including bioengineered devices, drug-eluting stents (scaffold to open occluded artery), and systemic drug therapy. Drugs currently approved for humans are very likely to find new uses in treatment of coronary disease. We are uniquely positioned to benefit the prevention and regression of coronary disease, which will improve health in the U.S.

描述（由申请人提供）：长期目标是确定冠状动脉平滑肌（CSM）腺苷A1和P2Y2受体信号传导机制，通过拮抗肾素-血管紧张素-醛固酮系统（RAAS）和运动训练来减少冠状动脉疾病（CAD）和代谢综合征（MetS）中的再狭窄。 RAAS 对 CSM A1 和 P2Y2 受体信号传导（调节激酶和下游炎症）的调节尚不清楚。尽管药物洗脱支架可减少再狭窄，但由于再狭窄并发症和支架附近 CAD（即支架周围 CAD）的进展，还需要进一步研究。我们之前资助的主要发现为该项目提供了重要的理由。奥萨博猪具有临床上显着的动脉粥样硬化（>50% 狭窄），我们首次在自然病变处植入支架，而不仅仅是气球损伤的健康动脉。我们从猪身上克隆了所有 4 种腺苷受体亚型（A1、A2A、A2B、A3），发现 A1 受体 (A1R) 在 CSM 中选择性表达，并在动脉粥样硬化和支架冠状动脉中增加。 A1R 的直接激活诱导 CSM 增殖，表明新的调节。我们克隆了 3 个猪 P2 核苷酸受体亚型，发现 P2Y2 亚型仅在患病的 CSM 中表达，并且在支架冠状动脉中选择性上调。总体假设：在 MetS 中，RAAS（主要是醛固酮）增加冠状动脉 A1R 和 P2Y2R 信号传导，这是 CSM 生长的关键信号。综合实验设计比较了瘦肉与 MetS Ossabaw RAAS 的拮抗作用。动脉粥样硬化病变被置入支架，然后在运动或不运动的情况下恢复。结果与更简单的体外器官培养进行比较。具体目标是测试 MetS 与瘦 Ossabaw 中的 6 个具体假设：1) 动脉粥样硬化、支架周围 CAD 和支架内狭窄在 MetS 中增加，并通过 RAAS 拮抗和运动减弱。血管内超声评估体内 CAD，光学成像和组织病理学评估体外 CAD。 2) MetS 增加 A1R 和 P2Y2R 的分子表达。将测定 5 个不同冠状动脉节段的蛋白质和 mRNA：健康节段、动脉粥样硬化节段、支架周围节段、支架内新内膜节段和支架内介质节段。 3) MetS 通过 ERK、JNK 和 AKT 激活增加 A1R 和 P2Y2R 的功能表达。磷酸化 ERK、JNK 和 AKT 的免疫印迹和免疫细胞化学；商店操作的 Ca 流入；炎症和氧化应激可量化功能激活。 4) 血管紧张素 II 和醛固酮直接、独立和协同地增加 A1R 和 P2Y2R 的分子和功能表达，血脂异常会增强这种作用。体内治疗包括几个猪组，与体外器官培养进行比较。 5) 运动会逆转 MetS 中 A1R 和 P2Y2R 分子和功能表达的增加。该目标评估更成熟的 CAD 和支架置入术。 6) 阻断 A1R 和 P2Y2R 可减轻过度扩张损伤和天然动脉粥样硬化中的支架内狭窄。瘦肉猪和 MetS 猪将接受药物和基因洗脱支架的体内冠状动脉支架置入术，以选择性阻断 A1R 和 P2Y2R。公共健康相关性：我们的研究涉及 Ossabaw 小型猪的临床高度相关的冠状动脉疾病，它完美地模拟了人类复杂的代谢综合征（“糖尿病前期”）和 2 型糖尿病。因此，我们的研究结果将促进新型治疗干预措施向人类的转化，包括生物工程设备、药物洗脱支架（打开闭塞动脉的支架）和全身药物治疗。目前批准用于人类的药物很可能在治疗冠心病方面找到新的用途。我们处于独特的地位，有利于预防和消退冠心病，这将改善美国的健康状况。