Cookson E J, Glover J
Department of Biochemistry, University of Liverpool.
J Endocrinol. 1987 Aug;114(2):191-8. doi: 10.1677/joe.0.1140191.
The disappearance of the thyroid hormone thyroxine (T4) from plasma in fully grown male Japanese quail can be described as a first order process with a rate constant of 0.178 +/- 0.013/h (mean +/- S.E.M., n = 8), which represents a half-life of 3.90 h. A small amount of [125I]T4 in relation to total circulating T4 was injected i.v. into Japanese quail and plasma samples were taken at appropriate time-intervals for the determination of residual plasma radioactivity. The rate of disappearance of [125I]T4 was subsequently equated to the turnover rate of the endogenous hormone. Previous methods were modified to overcome problems arising from possible disturbance of plasma T4 metabolism, recirculation of radiolabelled iodide, and to purify the [125I]T4 from the plasma samples. By using labelled T4 of very high specific activity, the amount of [125I]T4 administered was kept much smaller than has been used in previous studies on Japanese quail, thus limiting any interference with plasma T4 dynamics. To minimize any disturbance of plasma T4 metabolism, only four blood samples were taken, at three-hourly intervals after the injection of [125I]T4. The rapid turnover of T4 produced a large amount of labelled inorganic iodide, the re-entry of which into the plasma T4 pool was inhibited by s.c. administration of sodium thiocyanate 1 h before injection of [125I]T4. Assay of the true [125I]T4 turnover was significantly improved over that used in previous studies by purifying the [125I]T4 from the plasma samples chromatographically. The samples were applied to small Sephadex G-25 columns with sodium hydroxide (0.1 mol/l) as the eluant.(ABSTRACT TRUNCATED AT 250 WORDS)
在成年雄性日本鹌鹑中,甲状腺激素甲状腺素(T4)从血浆中的消失可描述为一级过程,速率常数为0.178±0.013/h(平均值±标准误,n = 8),半衰期为3.90小时。将少量相对于总循环T4的[125I]T4静脉注射到日本鹌鹑体内,并在适当的时间间隔采集血浆样本,以测定血浆中残留的放射性。随后将[125I]T4的消失速率等同于内源性激素的周转率。对先前的方法进行了改进,以克服血浆T4代谢可能受到干扰、放射性标记碘化物再循环以及从血浆样本中纯化[125I]T4等问题。通过使用比活性非常高的标记T4,所给予的[125I]T4量比先前对日本鹌鹑的研究中使用的量小得多,从而限制了对血浆T4动态的任何干扰。为了尽量减少对血浆T4代谢的干扰,在注射[125I]T4后,仅每隔三小时采集四个血样。T4的快速周转产生了大量标记的无机碘化物,在注射[125I]T4前1小时皮下注射硫氰酸钠可抑制其重新进入血浆T4池。通过色谱法从血浆样本中纯化[125I]T4,与先前研究中使用的方法相比,真正的[125I]T4周转率测定有了显著改进。将样本应用于以氢氧化钠(0.1 mol/l)为洗脱剂的小型Sephadex G - 25柱上。(摘要截断于250字)