A radiochemical assay for detection of leukotriene B4 production from isolated cells.
A radiochemical procedure for quantitating the effect of inhibitors of leukotriene B4 (LTB4) biosynthesis is described. Rat peritoneal cells were labeled with 3H-arachidonic acid and stimulated with the calcium ionophore A23187. 3H-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.
D015117:5,8,11,14-Eicosatetraynoic Acid / Q000494:pharmacology; D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D001202:Ascitic Fluid / Q000378:metabolism*; D000001:Calcimycin / Q000494:pharmacology; D002396:Catechols / Q000494:pharmacology; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D007213:Indomethacin / Q000494:pharmacology; D007700:Kinetics; D007975:Leukotriene B4 / Q000096:biosynthesis*; D008297:Male; D009637:Masoprocol; D051381:Rats; D011919:Rats, Inbred Strains; D014316:Tritium
D001095:Arachidonic Acids; D002396:Catechols; D014316:Tritium; D015117:5,8,11,14-Eicosatetraynoic Acid; D007975:Leukotriene B4; D016718:Arachidonic Acid; D000001:Calcimycin; D009637:Masoprocol; D007213:Indomethacin