Abstract
As lipid peroxides are produced from arachidonic acid(AA) by platelet lipoxygenase(PLO) and cyclo-oxygenase(PCO), a simple method was developed for estimation of these enzymic pathways by quantitating the thiobarbituric acid-reacting substance(TBARS) produced by the incubation of AA with human platelets. Thin-layer radiochromatographic analysis (J. Biol.Chem. 252:5871, 1977) of lipid products obtained by incubating [1-14C] AA with washed platelets under various conditions revealed that maximal amounts of PCO products were obtained by the incubation at 37°C within 1 min around pH 7.4, while those of PLO products within 10 min around pH 6.5. The incubation at 37°C of AA with aspirin-treated platelets at pH 7.4 for 1 min produced no TBARS, while that at pH 6.5 did TBARS as a function of the incubation time (up to 10 min). Based on these findings, PLO and PCO pathways were estimated by the determination of TBARS (J.Lab.Clin.Med. 75:283, 1970) obtained by incubating AA (0.2 mM, f.c.) either with aspirin-treated or with intact washed platelets (108) at 37°C as shown in the table. Normal values (M ± SD) expresse in terms of nmols of malonaldehyde were: PLO = 1. 76 ± 0 .36 (n=l7), PCO = 1.12 ± 0.42 (n=15). Alterations in these pathways were detecetd by this method in some patients with myeloproliferative disorders.
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