NMR STUDIES OF METAL IONS IN INTACT CELLS AND TISSUES

这项研究的主要目标仍然是阐明 细胞内金属离子（Na+、K+、游离Mg ~（2+）和游离Ca ~（2+））代谢 正常和病理状态。我们试图了解监管如何 的细胞内浓度的基本金属离子误入歧途， 健康问题，尤其是高血压和糖尿病。我们的主要研究 工具是核磁共振波谱。我们的实验室发挥了关键作用， 开发用于测量细胞内Na+的NMR方法（使用 膜损伤细胞外移位试剂，以区分 胞内和胞外离子）和游离Mg 2+（通过分析31 P NMR 细胞内ATP的化学位移），并用于同时测量 重金属和细胞内游离Ca 2+（通过氟化的19 F NMR） 引入细胞的探针分子）。细胞的选择和 我们研究的问题是基于可用性的标准， 生理意义它包括人类外周血细胞 （红细胞和血小板）、完整大鼠肾脏和心脏体内或 在体外灌注，并灌注大鼠主动脉，这是一个容易接近的 血管平滑肌细胞模型。以下具体目标将 （1）为了检验细胞内钾离子浓度与细胞内钾离子浓度之间的关系， 缺乏与高血压有关，并由于K+增加而出现 通过钙激活钾通道流出; 2）为了测试 一种假说认为，由于肾钠转运异常， 肾Na ~+：H ~+转运亢进伴血管离子 盐敏感性的变化，但不耐盐， 高血压：3）研究糖尿病对细胞内 [Na+]、[K+]、游离[Ca 2 +]、游离[Mg 2 +]和选定靶组织中的pH （4）在糖尿病大鼠模型中， 研究高血糖症引起糖尿病的潜在原因 相关的细胞内离子异常，特别是在血管 组织;和（5）调查糖尿病患者和 高血压肾和心肌缺血性损伤，以及 镁离子的保护作用。希望拟议的 高血压和糖尿病细胞内离子的研究将 最终导致更好的战略，他管理这些健康 紊乱

The primary goal of this research continues to be elucidation of intracellular metal ion (Na+, K+, free Mg2+ & free Ca2+) metabolism in normal and pathological states. We seek to understand how the regulation of intracellular concentrations of essential metal ions goes astray in health disorders, especially hypertension diabetes. Our main research tool is NMR spectroscopy. Our laboratory played a key role in the development of NMR methods for measuring intracellular Na+ (using a membrane-impairment extracellular shift reagent to differentiate between intra- and extracellular ions) and free Mg2+ (by analyzing 31P NMR chemical shifts of intracellular ATP), and for simultaneous measurement of heavy metals and intracellular free Ca2+ (by 19F NMR of fluorinated probe molecules introduced into the cell). The choice of cells and issues for our research is based on the criteria of availability and physiological significance. It includes human peripheral blood cells (erythrocytes and platelets), intact rat kidney and heart in-vivo or perfused in vitro, and perfused rat aorta which is a readily accessible model for vascular smooth muscle cells. The following specific aims will be pursued: (l) To test the hypothesis that intracellular potassium deficit is relevant in hypertension, and arises due to increased K+ efflux via calcium-activated potassium channels; 2) To test the hypothesis that an abnormality in renal sodium transport due to hyperactivity of renal Na+:H+ transport accompanies vascular ionic changes in salt-sensitive, but not in salt-resistant, essential hypertension: 3) To investigate the effects of diabetes on intracellular [Na+], [K+], free [Ca2+], free [Mg2+] and pH in elected target tissues (kidney, heart and vasculature) in diabetic rat models;(4) To investigate the potential ole of hyperglycemia in causing diabetes associated intracellular ionic abnormalities, especially in the vascular tissue; and (5) To investigate increased vulnerability of diabetic and hypertensive kidney and myocardium to ischemic damage, and the protective effect of magnesium ions. It is hoped that the proposed investigations of intracellular ions in hypertension and diabetes will eventually lead to better strategies for he management of these health disorders.