Yandle T G, Espiner E A, Nicholls M G, Duff H
J Clin Endocrinol Metab. 1986 Jul;63(1):72-9. doi: 10.1210/jcem-63-1-72.
A RIA for alpha-human atrial natriuretic peptide (alpha hANP) in plasma was developed and used to study the immunoreactive components secreted by the heart and circulating in peripheral venous plasma. The assay used [125I]diiodotyrosyl-alpha hANP, purified by high pressure liquid chromatography (HPLC), and a C-terminal-specific antiserum purchased from Peninsula Laboratories. Serial dilution curves of coronary sinus plasma samples were parallel with the standard curve, but significant nonparallelism was found in peripheral plasma samples of low immunoreactivity. When plasma was extracted using C-18 Sep-Pak cartridges, serial dilution curves from both coronary sinus and peripheral plasma samples were parallel to the standard curve. Although values for plasma samples assayed before and after extraction agreed closely (r = 0.99; n = 76), immunoreactive ANP in unextracted plasma was consistently greater (70-79 pmol/liter) than in extracts of plasma, suggesting non-specific interference by a component in plasma when assayed without extraction. Mean plasma immunoreactive ANP in 19 normal subjects consuming a normal salt intake was 14 +/- 1 (+/- SE) pmol/liter. In 5 normal men, increasing dietary sodium intake from 10 to 200 mmol sodium/day was associated with a 2-fold increment in ANP levels, and similar changes accompanied acute sodium loading using iv saline. Elevated values were found in patients with congestive heart failure (mean, 58 pmol/liter; range, 0-200; n = 9), chronic renal failure (mean, 118 pmol/liter; range, 30-290; n = 8), and primary aldosteronism (range, 32-90 pmol/liter; n = 3). HPLC and gel chromatographic analysis of the immunoreactive material found in coronary sinus plasma extracts showed that a large amount of the material eluted in the position of alpha hANP. A smaller quantity of immunoreactive material with a mol wt of about 1600 was also identified. Peripheral venous plasma extracts also contained several immunoreactive components, the largest amount of which corresponded to alpha hANP. The pattern of immunoreactive components in peripheral venous plasma, as identified by both gel chromotography and HPLC, was similar to that in coronary sinus plasma drawn during an active phase of hormone secretion. These findings indicate that the heart secretes alpha hANP or a closely similar peptide which is also present in peripheral venous plasma. Plasma immunoreactive ANP is responsive to sodium loading in normal man and is elevated in patients with hypervolemic disorders.
建立了一种放射免疫分析法（RIA）用于检测血浆中的α-人心房利钠肽（α-hANP），并用于研究心脏分泌并在外周静脉血浆中循环的免疫反应性成分。该分析方法使用经高压液相色谱（HPLC）纯化的[125I]二碘酪氨酸-α-hANP，以及从半岛实验室购买的C末端特异性抗血清。冠状窦血浆样本的系列稀释曲线与标准曲线平行，但在免疫反应性低的外周血浆样本中发现了明显的不平行性。当使用C-18 Sep-Pak柱提取血浆时，冠状窦和外周血浆样本的系列稀释曲线均与标准曲线平行。虽然提取前后测定的血浆样本值非常接近（r = 0.99；n = 76），但未提取血浆中的免疫反应性ANP始终高于血浆提取物中的（70 - 79 pmol/升），这表明在未提取情况下测定时血浆中的一种成分存在非特异性干扰。19名正常盐摄入的正常受试者的平均血浆免疫反应性ANP为14±1（±SE）pmol/升。在5名正常男性中，将饮食中钠摄入量从每天10 mmol增加到200 mmol与ANP水平增加2倍相关，静脉注射生理盐水进行急性钠负荷时也伴随类似变化。充血性心力衰竭患者（平均58 pmol/升；范围0 - 200；n = 9）、慢性肾衰竭患者（平均118 pmol/升；范围30 - 290；n = 8）和原发性醛固酮增多症患者（范围32 - 90 pmol/升；n = 3）中发现值升高。对冠状窦血浆提取物中发现的免疫反应性物质进行HPLC和凝胶色谱分析表明，大量物质在α-hANP位置洗脱。还鉴定出少量分子量约为1600的免疫反应性物质。外周静脉血浆提取物也含有几种免疫反应性成分，其中含量最多对应于α-hANP。通过凝胶色谱和HPLC鉴定的外周静脉血浆中免疫反应性成分的模式与激素分泌活跃期抽取的冠状窦血浆中的模式相似。这些发现表明心脏分泌α-hANP或一种与之密切相似的肽，其也存在于外周静脉血浆中。血浆免疫反应性ANP在正常人中对钠负荷有反应，在血容量过多性疾病患者中升高。
