Yamaji T, Ishibashi M, Takaku F
J Clin Invest. 1985 Oct;76(4):1705-9. doi: 10.1172/JCI112159.
To determine whether atrial natriuretic factor (ANF) is a circulating hormone in men, a radioimmunoassay suitable for the estimation of ANF in human plasma was developed and the nature of plasma ANF was characterized. Plasma ANF was extracted before radioimmunoassay by affinity chromatography on a column of ANF antibody-coupled agarose. When plasma ANF extract was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with the radioimmunoassay of the eluted gel slices for ANF, almost all of the ANF activities ran in the 3,000-mol-wt area, while three peaks of ANF were observed in human atrial tissue extract, molecular weights of which corresponded to 14,000, 6,000, and 3,000, respectively. Reversed-phase high performance liquid chromatography of atrial tissue extract resolved multiple forms of ANF. In contrast, one major peak was observed in human plasma extract, and its retention time coincided with that of synthetic human alpha-atrial natriuretic polypeptide. When 500 ml of 0.9% saline was infused into six healthy subjects over 45 min, plasma levels of ANF were unequivocally elevated. The mean plasma ANF concentrations rose from the baseline (23.0 +/- 2.5 pg/ml, mean +/- SEM, n = 6) to the peak (41.8 +/- 4.9 pg/ml, mean +/- SEM) at 75 min postinfusion. No significant change in plasma ANF, on the other hand, was found in the control group. These results suggest that ANF is a circulating hormone in men and is secreted in response to isotonic volume expansion.
为了确定心房利钠因子(ANF)在男性体内是否为一种循环激素,我们开发了一种适用于评估人血浆中ANF的放射免疫测定法,并对血浆ANF的性质进行了表征。在放射免疫测定之前,通过在ANF抗体偶联琼脂糖柱上进行亲和层析来提取血浆ANF。当通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析血浆ANF提取物,并对洗脱的凝胶切片进行ANF放射免疫测定时,几乎所有的ANF活性都出现在分子量为3000的区域,而在人心房组织提取物中观察到三个ANF峰,其分子量分别对应于14000、6000和3000。心房组织提取物的反相高效液相色谱法分离出多种形式的ANF。相比之下,在人血浆提取物中观察到一个主要峰,其保留时间与合成人α-心房利钠多肽的保留时间一致。当在45分钟内将500毫升0.9%的盐水注入6名健康受试者体内时,血浆ANF水平明显升高。输注后75分钟时,血浆ANF平均浓度从基线水平(23.0±2.5皮克/毫升,平均值±标准误,n = 6)升至峰值(41.8±4.9皮克/毫升,平均值±标准误)。另一方面,对照组中血浆ANF没有显著变化。这些结果表明,ANF在男性体内是一种循环激素,并且会因等渗容量扩张而分泌。