A novel assay of cell rubidium uptake using graphite furnace atomic absorption: application to rats on a magnesium-deficient diet.

A novel assay of cell rubidium uptake using graphite furnace atomic absorption: application to rats on a magnesium-deficient diet.
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使用石墨炉原子吸收来检测细胞铷吸收的新方法:应用于缺镁饮食的大鼠。

DOI:
10.1016/j.jnutbio.2005.01.005
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发表时间:
2005
期刊:
The Journal of nutritional biochemistry.
影响因子:
--
通讯作者:
Graves,StevenW
Graves,StevenW
中科院分区:
--
文献类型:
--
作者:
Zhen,Yueying;Franz,KayB;Graves,StevenW

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[Na,K] ATP酶或钠泵(SP)是普遍存在的膜阳离子转运系统。由于其在原发性高血压和白内障形成的病理生理学中的潜在参与,SP正在积极研究以详细说明其功能和控制。在本文中,我们描述了一种新的,非放射性的方法测量SP离子转运活性在完整的红细胞(RBC)使用石墨炉原子吸收测量铷离子(Rb)的摄取。该方法提供了与放射性技术相当的灵敏度,如通过用人红细胞(RBC)和哇巴因(一种已知的SP抑制剂)进行的实验所评估的,但是这种分析方法消除了使用其他Rb摄取测定方法所共有的放射性同位素。作为其更广泛实用性的证明,该测定用于评估膳食镁摄入对饮食控制大鼠RBC中SP介导的离子转运的影响。7周的镁缺乏(MgD)饮食的大鼠显示血清镁浓度显着降低,虽然水平保持在健康,镁充足的动物的参考区间的较低区域。红细胞Rb摄取显着减少镁限制动物的细胞,表明Rb摄取减少镁摄入量的敏感性,尽管血清水平下降在报告的正常范围内,和实用程序的Rb摄取测定在测量SP功能的生理变化。
The [Na,K]ATPase or sodium pump (SP) is a ubiquitous membrane cation transport system. Because of its potential participation in the pathophysiology of essential hypertension and cataract formation, the SP is under active investigation to detail its function and control. In this paper, we describe a novel, nonradioactive method of measuring SP ion transport activity in intact red blood cells (RBCs) using graphite furnace atomic absorption measurement of rubidium ion (Rb) uptake. This method provided sensitivity comparable to radioactive techniques, as assessed by experiments with human red blood cells (RBC) and ouabain, a known SP inhibitor, but this analytical approach eliminates the use of radioisotopes common to other Rb uptake assay methods. As a demonstration of its broader utility, the assay was used to assess the effects of dietary magnesium intake on SP-mediated ion transport in the RBCs of diet-controlled rats. Rats on 7 weeks of a magnesium-deficient (MgD) diet showed significant reductions in serum magnesium concentration, although levels remained in the lower region of the reference interval for healthy, magnesium replete animals. Red cell Rb uptake was significantly reduced in cells from the magnesium-restricted animals, demonstrating the sensitivity of Rb uptake to reduced magnesium intake, despite serum levels that fell within the reported normal range, and the utility of this Rb uptake assay in measuring physiological changes in SP function.
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