一种用于测定人中性粒细胞血小板活化因子合成的放射受体结合试验。

A radioreceptor binding assay for measurement of platelet-activating factor synthesis by human neutrophils.

作者信息

Paulson S K, Nicholson N S

机构信息

Department of Immunoinflammatory Diseases Research, Searle Research and Development, Skokie, IL 60077.

出版信息

J Immunol Methods. 1988 Jun 13;110(2):209-15. doi: 10.1016/0022-1759(88)90105-6.

DOI:10.1016/0022-1759(88)90105-6

PMID:2837517

Abstract

A new convenient method for the measurement of platelet-activating factor produced in vitro has been developed. This method involves incubation of neutrophils with stimuli, lipid extraction, purification of lipid extracts by normal phase high performance liquid chromatography (HPLC) and quantification of platelet-activating factor (PAF) by a competitive radioreceptor binding assay. The recovery of PAF from the extraction and purification procedures is 94.5 +/- 0.9%. The sensitivity of the assay is 10 pg/tube. Human neutrophils stimulated with 0, 0.25, 0.5, 1 and 2 microM A 23187 will produce ND, ND, 720, 840 and 900 pg of PAF, respectively (ND = not detectable). The values obtained for PAF produced by human neutrophils in the present assay are comparable to those obtained with the rabbit platelet aggregation assay.

摘要

已开发出一种用于体外测量血小板活化因子的新便捷方法。该方法包括将中性粒细胞与刺激物孵育、脂质提取、通过正相高效液相色谱（HPLC）纯化脂质提取物以及通过竞争性放射受体结合测定法定量血小板活化因子（PAF）。从提取和纯化过程中PAF的回收率为94.5±0.9%。该测定法的灵敏度为10 pg/管。用0、0.25、0.5、1和2 microM A 23187刺激的人中性粒细胞将分别产生ND、ND、720、840和900 pg的PAF（ND = 未检测到）。在本测定法中，人中性粒细胞产生的PAF值与兔血小板聚集测定法获得的值相当。