LHRH拮抗类似物过敏样反应的体外评价方法一组胺释放法

Evaluation Method of Anaphylactoid Activity of LHRH Antagonists—Histamine Release Test

本文主要报道了自行合成的九种LHRH拮抗类似物的组胺释放作用的评价方法。我们应用荧光分光光度法测定不同剂量LHRH拮抗类似物引起大鼠腹腔肥大细胞体外组胺释放量,以半数有效量(ED_(50))表示组胺释放作用的强弱。结果表明,九种LHRH拮抗类似物的ED_(50)在0.65±0.08μg/ml～36.80±5.68μg/ml范围内。本文还将此法检测得到的结果与使用皮肤过敏样反应几何法和分光光度法测得的结果进行分析,并比较了各种方法的优缺点。

The test of histamine release (HRT) from rat mast cell is one of the methods used to evaluate the anaphylactoid activity of LHRH antagonists. Using fluorometric technique, we determined ED_(50) of the histamine releasing activity of LHRH Antagonists. There were 9 LHRH antagonists synthesized by us and tested by this method. The range of ED_(50) was from 0.65 + 0.08 μg/ml to 36.80 + 5.68 Mg/ml. We selected a LHRH antagonist (N-AcNal (2)~1, D-pCl-Phe^2. D-3 Pal^(3,6), Arg^5, D-Ala) LHRH as a standard, which had been assayed and published in USA. Its ED_(50) was reported as 2.90 + 0.33 μg/ml in the literature. Our result was 3.50±0.38 μg/ml. We also compard the results of this method with those of cutaneous anaphylactoid test (CAT) by measuring either the diameters of lesion induced by the peptides or the quantities of Evan's blue locally by spectrophotometric method. It is concluded that the test of histamine release in vitro is sensitive, simple and reliable, and may be widely applied in the evaluation and screening of new LHRH antagonists.