Garthwaite T L, Hagen T C
J Clin Endocrinol Metab. 1978 Oct;47(4):885-8. doi: 10.1210/jcem-47-4-885.
An in vitro bioassay for plasma PRL-releasing factor-like activity has been developed. The method is a three-phase methanol extraction of plasma with extracts of 1.0 ml plasma adjusted to a final volume of 50 microliter. Single 50-microliter aliquots of extract were incubated in 1.0 ml Krebs-Ringer phosphate (KRP) buffer with one rat hemipituitary after a 1-h preincubation. Samples were obtained basally and 30 min after addition of the extract. During each set of incubations, a parallel series of hemipituitaries was incubated in KRP alone. The total nanograms of rat PRL released per mg pituitary tissue during the initial 30 min after preincubation was calculated for all studies. The mean quantity released in KRP alone was considered basal and was subtracted from values obtained during incubation with plasma extracts. The quantity remaining was considered PRL-releasing activity (PRA) of plasma, expressed as nanograms of rat PRL released per mg pituitary. The PRA in plasma from 13 patients with the amenorrhea-galactorrhea syndrome was 132 +/- 17 ng/mg pituitary (X +/- SE), which was significantly greater (P less than 0.001) than the PRA in plasma from eight matched controls [31 +/- 10 ng/mg pituitary (X +/- SE)]. The patients' individual PRL levels were elevated (range, 48-248 ng/ml), and when compared to the PRA in the samples, a highly significant (P less than 0.001) positive correlation evolved. These results indicate that a circulating PRL-releasing factor-like material present in normal plasma is higher in plasma from hyperprolactinemic patients in direct relationship to the PRL concentration. It is possible that this material is related to the pathogenesis of PRL-secreting pituitary disorders.
已开发出一种用于检测血浆催乳素释放因子样活性的体外生物测定法。该方法是对血浆进行三相甲醇提取,将1.0 ml血浆提取物调整至最终体积50微升。在1小时预孵育后,将50微升单份提取物等分试样与一个大鼠半垂体在1.0 ml Krebs-Ringer磷酸盐(KRP)缓冲液中孵育。在基础状态以及加入提取物后30分钟获取样本。在每组孵育过程中,将一系列平行的半垂体单独在KRP中孵育。对所有研究计算预孵育后最初30分钟内每毫克垂体组织释放的大鼠催乳素总纳克数。仅在KRP中释放的平均量被视为基础值,并从与血浆提取物孵育期间获得的值中减去。剩余的量被视为血浆的催乳素释放活性(PRA),以每毫克垂体释放的大鼠催乳素纳克数表示。13例闭经-溢乳综合征患者血浆中的PRA为132±17纳克/毫克垂体(X±SE),显著高于(P<0.001)8例匹配对照血浆中的PRA[31±10纳克/毫克垂体(X±SE)]。患者的个体催乳素水平升高(范围为48 - 248纳克/毫升),与样本中的PRA相比,呈现出高度显著的(P<0.001)正相关。这些结果表明,正常血浆中存在的一种循环催乳素释放因子样物质在高催乳素血症患者的血浆中含量更高,且与催乳素浓度呈直接关系。这种物质可能与分泌催乳素的垂体疾病的发病机制有关。