Cellular and Molecular Mechanisms of Coronary Endothelial Dysfunction in Diabetes

糖尿病冠状动脉内皮功能障碍的细胞和分子机制

基本信息

批准号：
8883693
负责人：
Ayako Makino
金额：
$ 37.68万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-03 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8883693
关键词：
20 year old Abnormal Endothelial Cell Accounting Apoptosis Apoptotic Attenuated Binding Blood Glucose Blood Vessels Blood capillaries Blood flow Ca(2+)-Transporting ATPase Calcium-Activated Potassium Channel Cardiac Myocytes Categories Cause of Death Cell Death Cell membrane Cells Communication Complication Coronary Coronary artery Coronary heart disease Coupled Data Defect Diabetes Mellitus Diabetic Angiopathies Diabetic Microangiopathies Diabetic mouse Disease Down-Regulation Endoplasmic Reticulum Endothelial Cells Endothelium Functional disorder Gap Junctions Generations Glucose Grant Heart Hyperglycemia Hyperlipidemia Incidence Individual Injury Ischemic Stroke Lead Left ventricular structure Mediating Membrane Mitochondria Molecular Morbidity - disease rate Myocardial Ischemia Newly Diagnosed Nitric Oxide Synthase Nutrient Organelles Oxygen Play Post-Translational Protein Processing Potassium Channel Proteins Pump Regulation Relaxation Role STIM1 gene Signal Transduction Site Stroke Testing Transportation United States Vascular Endothelium-Dependent Relaxation Vasodilation Vasodilator Agents base capillary cell type connexin 40 density design diabetic diabetic patient endothelial dysfunction extracellular gap junction channel insight mortality neuron loss novel therapeutic intervention overexpression protein expression sarcoplasmic reticulum calcium ATPase sensor vascular endothelial dysfunction

项目摘要

DESCRIPTION (provided by applicant): Coronary heart disease and ischemic stroke are the main causes of morbidity and mortality in diabetic patients. Endothelial injury and dysfunction are common starting points for diabetic angiopathy. It is thus important to investigate the cellula and molecular mechanisms involved in coronary vascular endothelial dysfunction in diabetes. Endothelial function is regulated by changes in cytosolic Ca2+ concentration ([Ca2+]cyt) in endothelial cells (ECs). [Ca2+]cyt is controlled by the Ca2+ mobilization from intracellular stores coupled to Ca2+ influx from the external medium. In ECs, the endoplasmic reticulum (ER) accounts for approximately 75% of the total intracellular Ca2+ stores and the [Ca2+] in the ER ([Ca2+]ER) determines in great part the generation of important Ca2+ signals. The objective of this study is to examine whether [Ca2+]ER is altered in diabetic ECs and how abnormal [Ca2+]ER leads to vascular endothelial dysfunction in diabetic heart. In Aim 1, we will examine whether and how the rise in [Ca2+]cyt due to Ca2+ release from the ER (indicative of the level of [Ca2+]ER) and [Ca2+]ER are attenuated in coronary ECs isolated from diabetic mice. The ER membrane constitutes Ca2+ pumps (sarco/endoplasmic reticulum Ca2+ ATPase (SERCA)) and several classes of intracellular Ca2+ releasing channels. Recently, stromal interaction molecule (STIM) was identified as an essential protein for the store-operated Ca2+ entry (SOCE) and a contributor to the Ca2+ refilling into the ER by interacting with SERCA on the ER membrane. In Aim 2, we will investigate the potential role of STIM1 in decreased [Ca2+]ER in diabetic coronary ECs. Our preliminary data show that SERCA3 and STIM1 protein expressions are markedly decreased in diabetic coronary ECs and STIM1 overexpression in diabetic coronary ECs significantly increases Ca2+ leak from the ER. In addition, Ca2+ transported to mitochondria from the ER plays a crucial role in cell apoptosis; increased mitochondrial Ca2+ ([Ca2+]mit) can trigger mitochondrial fragmentation, loss of mitochondrial integrity and cell death in many cell types. Our results show that [Ca2+]mit is significantly increased in coronary ECs in diabetes. In Aim 3, we will define whether Ca2+ transportation from the ER to mitochondria is increased in coronary ECs in diabetes and examine the role of the Ca2+ communication between the ER and mitochondria in endothelial dysfunction in diabetic heart. To examine Ca2+ handling by the ER in diabetic coronary ECs will further our understanding of pathophysiological aspects of coronary vascular complications in diabetes. Completion of this study will provide important insights into developing new therapeutic interventions for cardiac ischemia in diabetes.

描述（申请人提供）：冠心病和缺血性中风是糖尿病患者发病和死亡的主要原因。内皮损伤和功能障碍是糖尿病血管病的常见起点。因此，研究糖尿病冠状血管内皮功能障碍的细胞和分子机制非常重要。内皮功能通过内皮细胞 (EC) 中胞质 Ca2+ 浓度 ([Ca2+]cyt) 的变化来调节。 [Ca2+]cyt 受细胞内储存的 Ca2+ 动员控制与来自外部介质的 Ca2+ 流入耦合。在 EC 中，内质网 (ER) 约占细胞内总 Ca2+ 储存的 75%，ER 中的 [Ca2+] ([Ca2+]ER) 在很大程度上决定了重要 Ca2+ 信号的产生。本研究的目的是检查糖尿病 EC 中 [Ca2+]ER 是否发生改变以及异常 [Ca2+]ER 如何导致糖尿病心脏血管内皮功能障碍。在目标 1 中，我们将检查从糖尿病小鼠分离的冠状动脉 EC 中，由于 ER 中 Ca2+ 释放（指示 [Ca2+]ER 水平）和 [Ca2+]ER 导致的 [Ca2+]cyt 增加是否以及如何减弱。内质网膜由 Ca2+ 泵（肌浆/内质网 Ca2+ ATP 酶 (SERCA)）和几类细胞内 Ca2+ 释放通道组成。最近，基质相互作用分子 (STIM) 被确定为钙库操纵的 Ca2+ 进入 (SOCE) 的必需蛋白，并且通过与 ER 膜上的 SERCA 相互作用，促进 Ca2+ 再填充到 ER 中。在目标 2 中，我们将研究 STIM1 在糖尿病冠状动脉 EC 中 [Ca2+]ER 降低中的潜在作用。我们的初步数据表明，糖尿病冠状动脉 EC 中 SERCA3 和 STIM1 蛋白表达显着降低，糖尿病冠状 EC 中 STIM1 过表达显着增加 ER 的 Ca2+ 渗漏。此外，从内质网转运到线粒体的Ca2+在细胞凋亡中起着至关重要的作用。线粒体 Ca2+ ([Ca2+]mit) 增加可引发多种细胞类型的线粒体断裂、线粒体完整性丧失和细胞死亡。我们的结果表明，糖尿病患者冠状动脉 EC 中的 [Ca2+]mit 显着增加。在目标 3 中，我们将确定糖尿病患者冠状动脉 EC 中从 ER 到线粒体的 Ca2+ 转运是否增加，并检查 ER 和线粒体之间的 Ca2+ 通讯在糖尿病心脏内皮功能障碍中的作用。检查糖尿病冠状动脉 EC 中 ER 对 Ca2+ 的处理将进一步加深我们对糖尿病冠状血管并发症病理生理学方面的理解。这项研究的完成将为开发糖尿病心脏缺血的新治疗干预措施提供重要见解。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Hypoxia-induced pulmonary hypertension in type 2 diabetic mice.

DOI：
10.1086/690206
发表时间：
2017-03
期刊：
Pulmonary circulation
影响因子：
2.6
作者：
Pan M;Han Y;Si R;Guo R;Desai A;Makino A
通讯作者：
Makino A

VDAC: old protein with new roles in diabetes.

VDAC：旧蛋白质在糖尿病中发挥新作用。

DOI：
10.1152/ajpcell.00087.2012
发表时间：
2012
期刊：
American journal of physiology. Cell physiology
影响因子：
0
作者：
Sasaki,Koh;Donthamsetty,Reshma;Heldak,Michael;Cho,Young-Eun;Scott,BrianT;Makino,Ayako
通讯作者：
Makino,Ayako