Direct measurement of human plasma corticotropin-releasing hormone by "two-site" immunoradiometric assay.
Direct measurement of human plasma corticotropin-releasing hormone by "two-site" immunoradiometric assay.
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通过“双位点”免疫放射测定法直接测量人血浆促肾上腺皮质激素释放激素。
DOI：
10.1210/jcem-64-5-1047
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
P. Lowry
中科院分区：
文献类型：
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作者：
E. Linton;C. McLean;A. C. Kruseman;F. Tilders;E. V. Veen;P. Lowry
描述了一种“两点”免疫放射测定法（IRMA），它允许直接估计血浆中的人类CRH （hCRH）。使用该IRMA，正常受试者的CRH基础水平为2-28 pg/mL[平均值，15 +/- 7 (+/- SD) pg/mL；N = 58]。男性和女性的价值观相似。Cushing’s syndrome、Addison’s disease和Nelson’s syndrome患者的血浆CRH值也在此范围内，这些患者的血浆CRH与ACTH水平无相关性。近期孕妇血浆CRH水平升高[1462 +/- 752 (+/- SD) pg/mL；n = 55]，该crh样免疫反应性稀释曲线与IRMA标准曲线平行。从母体血浆中免疫吸附后，该CRH样物质经反相高效液相色谱洗脱，保留时间与合成CRH相同，在灌注垂体前叶细胞生物测定中与合成肽具有相同的生物活性。在Wistar大鼠中，即使在肾上腺切除术和随后的乙醚应激后，也未检测到循环CRH。新鲜人血浆降解合成hCRH的速度相对较慢；在37℃条件下孵育1小时后，加入的CRH仍保留65%。热处理（54℃；1小时）、冷处理（4℃；4小时）或冷冻和解冻均抑制降解。使用新鲜大鼠血浆时，合成大鼠CRH丢失更快；在相同的条件下，只有20%的CRH保持不变。无法测量外周大鼠血浆中的CRH可能是由于活性CRH降解酶的存在，这些酶将CRH分子分解成CRH IRMA无法识别的形式。
A "two-site" immunoradiometric assay (IRMA) which allows the direct estimation of human CRH (hCRH) in plasma is described. Using this IRMA, basal levels of CRH in normal subjects ranged from 2-28 pg/mL [mean, 15 +/- 7 (+/- SD) pg/mL; n = 58]. Values in men and women were similar. Plasma CRH values within this range were also found in patients with Cushing's syndrome, Addison's disease, and Nelson's syndrome, with no correlation between plasma CRH and ACTH levels in these patients. Elevated plasma CRH levels were found in pregnant women near term [1462 +/- 752 (+/- SD) pg/mL; n = 55], and the dilution curve of this CRH-like immunoreactivity paralleled the IRMA standard curve. After its immunoadsorption from maternal plasma, this CRH-like material eluted on reverse phase high performance liquid chromatography with a retention time identical to that of synthetic CRH and had equipotent bioactivity with the synthetic peptide in the perfused anterior pituitary cell bioassay. Circulating CRH was not detected in Wistar rats, even after adrenalectomy and subsequent ether stress. Synthetic hCRH was degraded by fresh human plasma relatively slowly; 65% of added CRH remained after 1 h of incubation at 37 C. Degradation was inhibited by heat treatment (54 C; 1 h), cold treatment (4 C; 4 h), or freezing and thawing. Loss of synthetic rat CRH occurred more rapidly when fresh rat plasma was used; only 20% of added CRH remained under the same conditions. The inability to measure CRH in peripheral rat plasma may be due to the presence of active CRH-degrading enzymes which fragment the CRH molecule into forms not recognized by the CRH IRMA.
DOI：
10.1073/pnas.80.15.4851
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
RIVIER, J;SPIESS, J;VALE, W
通讯作者：
VALE, W