Requirement of free arachidonic acid for leukotriene B4 biosynthesis by 12-hydroperoxyeicosatetraenoic acid-stimulated neutrophils |
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| Authors: | K Kanaji M Okuma T Sugiyama S Sensaki F Ushikubi H Uchino |
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| Affiliation: | 1. Reprogenetics UK, Institute of Reproductive Sciences, Oxford Business Park North, Oxford OX4 2HW, UK;2. CEINGE-Biotecnologie Avanzate s.c.a r.l., Via Gaetano Salvatore, 486, 80145 Naples, Italy;3. Department of Neuroscience, Reproductive Medicine, Odontostomatology, University of Naples “Federico II”, Via S Pansini 5, 80131 Naples, Italy;4. Department of Medicine and Surgery, Section of Endocrinology, University of Naples Federico II, Via S Pansini 5, 80131 Naples, Italy;5. Nuffield Department of Obstetrics and Gynaecology, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DU, UK;1. Laboratory of Developmental Biology and Genetics (LDB & G), Department of Molecular Biology, Faculty of Science, University of South Bohemia, Branišovská 31, České Budějovice 37005, Czech Republic;2. Institute of Entomology, Biology Centre of the Czech Academy of Sciences, Branišovská 31, České Budějovice 37005, Czech Republic;1. GENERA centers for reproductive medicine, via G de Notaris 2/b, Rome, Italy;2. University of Rome, Sapienza, Dipartimento di Scienze Anatomiche, Istologiche, Medico-Legali e dell''Apparato Locomotore, Sezione Istologia ed Embriologia Medica, via A. Scarpa 16, Rome, Italy;3. GENETYX Molecular Genetics Laboratory, via Fermi 1, Marostica, Italy |
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| Abstract: | Stimulation of human neutrophils with 12-hydroperoxyeicosatetraenoic acid (12-HPETE) led to formation of 5S, 12S-dihydroxyeicosatetraenoic acid (DiHETE), but leukotriene B4 (LTB4) or 5-hydroxyeicosatetraenoic acid (5-HETE) was not detectable by reversed-phase high-performance liquid chromatography analysis. N-formylmethionylleucylphenylalanine (FMLP) induced the additional synthesis of small amounts of LTB4 in 12-HPETE-stimulated neutrophils. The addition of arachidonic acid greatly increased the synthesis of LTB4 and 5-HETE by neutrophils incubated with 12-HPETE. In experiments using [1-14C]arachidonate-labeled neutrophils, little radioactivity was released by 12-HPETE alone or by 12-HPETE plus FMLP, while several radiolabeled compounds, including LTB4 and 5-HETE, were released by A23187. These findings demonstrate that LTB4 biosynthesis by 12-HPETE-stimulated neutrophils requires free arachidonic acid which may be endogenous or exogenous. |
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