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血浆血管加压素细胞化学测定法的开发:应用于水负荷正常男性的研究。

Development of a cytochemical assay for plasma vasopressin: application to studies on water loading normal man.

作者信息

Baylis P H, Pippard C, Gill G V, Burd J

出版信息

Clin Endocrinol (Oxf). 1986 Apr;24(4):383-93. doi: 10.1111/j.1365-2265.1986.tb01642.x.

Abstract

A cytochemical assay has been developed to measure human plasma arginine vasopressin. It is based on the stimulation of Na+-K+, ATPase activity located in the outer medulla of the rat kidney, and is capable of detecting very low plasma arginine vasopressin concentrations, limit of detection 0.01 pmol/l. Specificity for vasopressin stimulation of the enzyme is conferred on the assay by the use of specific vasopressin antiserum. Index of precision of the assay is 0.21. Degradation of arginine vasopressin in plasma in inhibited by phenanthroline. Samples may be stored up to 8 weeks at -70 degrees C. Intra- and inter-assay coefficients of variation were 22% (n = 8) and 104% (n = 12), respectively. A sustained water load in eight healthy male adults caused a fall in plasma osmolality from a basal of 286.5 +/- 2.0 (mean +/- SEM) to 279.2 +/- 2.4 mmol/kg after the load (P less than 0.001), which was associated with a reduction in urine osmolality from 867 +/- 54 to 69 +/- 3 mmol/kg. Plasma immunoreactive arginine vasopressin fell from 1.3 +/- 0.3 pmol/l to become undetectable (less than 0.3 pmol/l), but plasma cytochemical arginine vasopressin decreased from 0.96 +/- 0.14 to 0.07 +/- 0.02 pmol/l. There was a curvilinear relationship between plasma osmolality and plasma cytochemical arginine vasopressin, which militated against the concept of an osmotic threshold for vasopressin release.

摘要

已开发出一种细胞化学测定法来测量人血浆中的精氨酸加压素。该方法基于对大鼠肾外髓质中Na+-K+、ATP酶活性的刺激,能够检测到极低的血浆精氨酸加压素浓度,检测限为0.01 pmol/l。通过使用特异性精氨酸加压素抗血清,该测定法赋予了对酶的加压素刺激的特异性。该测定法的精密度指数为0.21。邻菲罗啉可抑制血浆中精氨酸加压素的降解。样本可在-70℃下储存长达8周。批内和批间变异系数分别为22%(n = 8)和104%(n = 12)。八名健康男性成年人持续饮水负荷后,血浆渗透压从基础值286.5±2.0(平均值±标准误)降至负荷后279.2±2.4 mmol/kg(P<0.001),同时尿渗透压从867±54降至69±3 mmol/kg。血浆免疫反应性精氨酸加压素从1.3±0.3 pmol/l降至不可检测水平(<0.3 pmol/l),但血浆细胞化学精氨酸加压素从0.96±0.14降至0.07±0.02 pmol/l。血浆渗透压与血浆细胞化学精氨酸加压素之间存在曲线关系,这对加压素释放的渗透阈值概念提出了质疑。

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