INDIVIDUAL HUMAN PERIPHERAL MONONUCLEAR CELL MAGNESIUM QUANTITATION

个体人类外周单核细胞镁定量

• 批准号: 4705640
• 负责人: G R HOOK
• 金额: --
• 依托单位: DIVISION OF RESEARCH SERVICES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/4705640
• 关键词: X ray spectrometry; blood chemistry; electron probe spectrometry; human tissue; lymphocyte; magnesium; monocyte

Total body magnesium (Mg) is inversely correlated with heart disease, atherosclerosis and hypertension. Serum Mg is a poor indicator of total body Mg but peripheral blood mononuclear cell Mg may be a good indicator. Peripheral blood mononuclear cells are a heterogenous population of lymphocytes and monocytes, with monocytes being twice the volume of lymphocytes. To evaluate mononuclear cell Mg, the mean cell Mg of the population, the individual cell Mg distribution within the population and the relationships between cell Mg and cell size must be determined. Whereas atomic absorption spectroscopy (AAS) methods have been developed to measure the mean mononuclear cell Mg, methods for measuring the Mg of individual mononuclear cells have not been described. We have developed a method for the quantification of Mg in individual whole mononuclear cells by electron probe X-ray microanalysis (EPMA). Whole blood is anticoagulated with heparin, washed with buffered saline glucose and ultracentrifuged in a discontinuous stractan density gradient. Isolated mononuclear cells are washed with osoosmotic, pH neutral ammonium nitrate, spray deposited on thin film supports, air dried and carbon coated for EPMA. The Mg concentration is determined by the peak-to-local continuum X-ray intensity ratio. Glass microsphere standards obtained from the National Bureau of Standards were used to calibrate the analysis. The mean cell Mg concentrations determined by AAS and EPMA were not significantly different. The coefficient of variation for replicate analysis of the standard by EPMA was 10%. The net variation in Mg concentration among individual cells was 20% and the distribution was symetrical. The Mg concentration was independent of cell size but the cell Mg content (fg/cell) increased with cell size.

全身镁（Mg）与心脏病呈负相关， 动脉粥样硬化和高血压。 血清镁是一个很差的指标， 外周血单个核细胞Mg可能是一个较好的指标。 外周血单个核细胞是一个异质性群体， 淋巴细胞和单核细胞，单核细胞的体积是 淋巴细胞 为了评估单核细胞Mg，使用了单核细胞的平均细胞Mg。 群体内的单个细胞Mg分布，以及 必须确定晶胞Mg和晶胞尺寸之间的关系。 而原子吸收光谱（AAS）方法已经发展到 测量平均单核细胞Mg，测量 单个的单核细胞没有被描述。 我们已经开发出一种 单个完整单核细胞中镁的定量方法 电子探针X射线微区分析（EPMA）。 全血 用肝素抗凝，用缓冲盐水葡萄糖洗涤， 在一个不连续的stractan密度梯度中超密集。 分离 单核细胞用等渗的、pH中性的硝酸铵洗涤， 喷涂沉积在薄膜载体上，空气干燥并涂碳形成 电子探针。 Mg浓度由峰-局域连续谱决定 X射线强度比。 玻璃微球标准品获自 使用美国国家标准局校准分析。 平均 AAS和EPMA测定的细胞镁浓度差异不显著 不同. 重复分析的变异系数 EPMA标准品为10%。 Mg浓度的净变化 单个细胞占20%，分布对称。 的Mg 浓度与细胞大小无关，但细胞镁含量 (fg/cell）随细胞大小而增加。