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+、游离镁离子和游离钙离子)代谢 正常和病理状态。我们试图理解这一规定是如何 细胞内必需金属离子的浓度迷失在 健康障碍，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。我们的实验室在这一过程中发挥了关键作用 核磁共振法测定细胞内Na+的研究进展 膜损伤细胞外移位试剂鉴别 胞内和胞外离子)和游离镁离子(通过分析31P核磁共振 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钙的含量(通过~(19)F-核磁共振氟化 引入细胞的探针分子)。选择单元格和 我们研究的问题是基于可获得性和 生理意义。它包括人类外周血细胞 (红细胞和血小板)，体内完整的大鼠肾脏和心脏或 体外灌流，并灌流大鼠主动脉，这是一种容易接近的 血管平滑肌细胞模型。以下具体目标将 被追查：(L)检验细胞内钾离子 缺乏症与高血压有关，是由于K+增加引起的 通过钙激活的钾通道外流；2)测试 假设肾脏钠转运异常是由于 肾Na~+：H~+转运亢进伴随血管离子 盐敏感的变化，但不是耐盐的变化，是必要的 高血压：3)研究糖尿病对细胞内的影响 选定靶组织中的[Na+]、[K+]、游离[Ca2+]、游离[Mg2+]和pH 糖尿病大鼠模型(肾、心、血管)； 研究高血糖导致糖尿病的潜在原因 相关的细胞内离子异常，特别是在血管内 组织；以及(5)调查糖尿病和糖尿病易感性增加 高血压对肾脏和心肌的缺血性损害，以及 镁离子的保护作用。希望建议的 高血压和糖尿病患者细胞内离子的研究 最终导致更好的策略来管理这些健康 精神错乱。

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.