Coronary ion channels, gender and vasoreactivity

冠状动脉离子通道、性别和血管反应性

基本信息

批准号：
7440355
负责人：
DOUGLAS K BOWLES
金额：
$ 34.85万
依托单位：
UNIVERSITY OF MISSOURI-COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-01 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7440355
关键词：
Acute Attenuated Blood Vessels Blood flow Buffers Calcium Calcium-Activated Potassium Channel Cardiology Cells Clinical Clinical Trials Condition Coronary Coronary Artery Vasospasm Coronary heart disease Coupled Coupling Culture Techniques Cytophotometry Disease Electrophysiology (science)Estradiol Estrogens Family suidae Female Functional disorder Gender Gender Role Goals Gonadal Steroid Hormones Health Hormones Human Immunoblotting In Vitro Injury Ion Channel Isometric Exercise Knowledge Modeling Molecular Numbers Organ Physiological Physiology Potassium Channel Probability Protein Kinase C Regulation Research Reverse Transcriptase Polymerase Chain Reaction Role Severities Sex Characteristics Smooth Muscle Smooth Muscle Myocytes Techniques Testosterone Translations Vascular Diseases Vascular Smooth Muscle clinically relevant density in vivo insight large-conductance calcium-activated potassium channels male novel patch clamp prospective response sex voltage voltage clamp voltage gated channel

项目摘要

DESCRIPTION (provided by applicant): Recent prospective clinical trials have underscored our lack of knowledge with regard to the effect of gender and sex hormones on the coronary vasculature, especially at the cellular/molecular level. Sex hormones exert profound influence on vascular smooth muscle physiology, including proliferation, ion channel activity and channel expression. Coronary smooth muscle (CSM) ion channel activity is central to both regulation of coronary blood flow and progression of vascular disease. We have demonstrated that CSM calcium and potassium channel activities are strongly influenced by gender. This study will determine the mechanism for gender-specific differences in CSM voltage-gated calcium (VGCC) and K channel activity and the consequent effects on intracellular calcium (Cai) regulation and vasoreactivity. The overall hypothesis is that coronary arterial reactivity in males is greater due to a testosterone (TST)-dependent increase in VGCC synthesis and PKC-dependent activity resulting in an enhanced propensity for coronary vasospasm (CVS). Both in vivo (intact, gonadectomized and hormone-replaced swine) and in vitro techniques will be used to determine the role of gender, TST and estrogen (E2) in regulating CSM ion channel activity and coronary arterial reactivity. Aim 1 will determine the mechanism of gender and sex hormone induced changes in VGCC activity and expression using electrophysiology, immunoblot and RT-PCR. As both vasoreactivity and VGCC activity are modulated by PKC, gender and sex hormone effects on PKC regulation of VGCC will also be examined. As the effect of VGCC activity on Cai and contraction is modulated via coupling to K channels and cellular calcium buffering, Aim 2 will determine the effect of sex-specific differences in VGCC activity on Ca-activated (BK) and voltage-dependent K current (Kv) and Cai using simultaneous voltage clamp and microfluorometry. The effect of sex-specific differences in VGCC activity on macro- and microvascular reactivity under both physiological and pathophysiological conditions will be assessed in vitro and in vivo in Aim 3. A model of CVS will be used to assess the role of sex-specific differences in VGCC activity on the progression and severity of vascular disease. The goal of this research is to determine gender-related cellular and molecular differences in CSM as they relate to gender differences in propensity for coronary disease.

描述（由申请人提供）：最近的前瞻性临床试验强调了我们对性别和性激素对冠状动脉脉管系统的影响的知识，尤其是在细胞/分子水平上。性激素对血管平滑肌生理产生深远的影响，包括增殖，离子通道活性和通道表达。冠状动脉平滑肌（CSM）离子通道活性既是调节冠状动脉流动和血管疾病进展的核心。我们已经证明，CSM钙和钾通道活性受性别的强烈影响。这项研究将确定CSM电压门控钙（VGCC）和K通道活性的性别特异性差异的机制，以及对细胞内钙（CAI）调节和血管反应性的影响。总体假设是，由于睾丸激素（TST）依赖性VGCC合成和PKC依赖性活性，雄性冠状动脉反应性更大，从而增强了冠状动脉血管痉挛倾向（CVS）。体内（完整，性腺切除和激素重取的猪）以及体外技术都将用于确定性别，TST和雌激素（E2）在调节CSM离子通道活性和冠状动脉动脉反应性中的作用。 AIM 1将通过电生理学，免疫印迹和RT-PCR来确定性别和性激素诱导VGCC活性和表达的变化。由于血管反应性和VGCC活性都受到PKC的调节，因此还将检查性别和性激素对VGCC PKC调控的影响。由于VGCC活性对CAI和收缩的影响通过耦合与K通道和细胞钙缓冲进行调节，AIM 2将确定VGCC活性对CA激活（BK）和电压依赖性K电流（KV）和CAI的影响，并使用同时使用同时的电压挡板和微荧光和微荧光。在生理和病理生理状况下，VGCC活性中性别特异性差异对宏观和微血管反应性的影响将在AIM 3中评估和体内评估。CVS的模型将用于评估性别特异性差异在VGCC活性在血管疾病进展和严重性血管疾病中的作用。这项研究的目的是确定CSM中与性别相关的细胞和分子差异，因为它们与冠状动脉疾病倾向的性别差异有关。