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A radiochemical assay for detection of leukotriene B4 production from isolated cells

Journal Article · · J. Pharmacol. Methods; (United States)
; ;

A radiochemical procedure for quantitating the effect of inhibitors of leukotriene B4 (LTB4) biosynthesis is described. Rat peritoneal cells were labeled with /sup 3/H-arachidonic acid and stimulated with the calcium ionophore A23187. /sup 3/H-LTB4 was isolated by processing on C18-Sep Pak cartridges followed by reverse-phase high pressure liquid chromatography (HPLC). The identity of this product, as well as other arachidonic acid metabolites, was verified by using silicic acid column chromatography followed by straight-phase HPLC and thin layer chromatography. Using this assay, LTB4 release by ionophore A23187 has been shown to be both time- and concentration-dependent. Indomethacin enhanced, while NDGA and ETYA inhibited, the A23187-induced production of LTB4. This procedure is both simple and direct and is capable of assessing the ability of novel compounds to alter LTB4 production.

Research Organization:
Department of Pharmacology, Stuart Pharmaceuticals, Wilmington, Delaware
OSTI ID:
6747422
Journal Information:
J. Pharmacol. Methods; (United States), Journal Name: J. Pharmacol. Methods; (United States) Vol. 11:2; ISSN JPMED
Country of Publication:
United States
Language:
English

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Related Subjects

550501* -- Metabolism-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH METALS
ANIMALS
ARACHIDONIC ACID
BIOSYNTHESIS
CALCIUM
CARBOXYLIC ACIDS
CHROMATOGRAPHY
DOSE-RESPONSE RELATIONSHIPS
ELEMENTS
ENZYME INHIBITORS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
MEMBRANE TRANSPORT
METALS
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PROSTAGLANDINS
RATS
RODENTS
SEPARATION PROCESSES
SYNTHESIS
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES