DEVELOPMENT OF EPR CATHETER PROBES FOR IN VIVO STUDIES

用于体内研究的 EPR 导管探针的开发

• 批准号: 6206515
• 负责人: TADEUSZ WALCZAK
• 金额: $ 1.22万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6206515
• 关键词: Mammalia; biological products; biomaterials; biomedical resource; environmental toxicology; enzymes; hematology

Although nitric oxide (NO) is a central mediator during endotoxin-induced sepsis, direct detection of tissue NO in vivo, has until recently been difficult, and techniques have relied on indirect measurement of byproducts in blood or invasive technology. We have utilized electron paramagnetic resonance (EPR) in conjunction with the spin-trapping technique to detect NO directly, and non-invasively, from the tissue of septic mice. Relative signal intensity arising from NO complexed with iron and diethyldithiocarbamate (DETC) measured directly from the liver and kidney of mice given endotoxin was maximal at 6 hours post endotoxin. The quality of the EPR signal obtained (high signal to noise ratio of 15:1) using our experimental set-up and L-band EPR hardware was such that we were able to establish a time course of NO production in tissue following endotoxin, and measurement of NO from other organs (kidney and spleen). This technique was extended to experiments in which we first implanted an oxygen sensitive material (gloxy) into the liver of mice, and then monitored NO production following endotoxin. Due to the fact that the EPR spectrum from gloxy and that of NO-Fe-(DETC)2 do not overlap, we were able to monitor NO production and pO2 simulatneously in tissue, in real time.

虽然一氧化氮（NO）是一个中心介质， 内毒素诱导的脓毒症，直接检测体内组织NO， 直到最近才变得困难，技术依赖于间接的 测量血液中的副产物或侵入性技术。 我们有 利用电子顺磁共振（EPR）结合 利用自旋捕获技术直接、非侵入性地检测NO， 从脓毒症小鼠的组织中分离出来 出现的相对信号强度 从NO与铁和二乙基二硫代氨基甲酸盐（DETC）络合， 直接从肝脏和肾脏中提取的内毒素最多 在内毒素后6小时。 所获得的EPR信号的质量 (high信噪比为15：1）使用我们的实验装置， L波段EPR的硬件使得我们能够确定 内毒素作用下组织中NO产生的过程及测定 其他器官（肾脏和脾脏）的NO。 该技术 扩展到我们第一次植入氧气 敏感材料（gloxy）进入小鼠肝脏，然后监测 内毒素后无生成。 由于EPR 来自gloxy和NO-Fe-（DETC）2光谱不重叠，我们 能够同时监测组织中NO的产生和pO 2， 真实的时间。