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Related Experiment Videos

16(R)-hydroxy-5,8,11,14-eicosatetraenoic acid, a new arachidonate metabolite in human polymorphonuclear leukocytes.

M M Bednar1, C E Gross, M K Balazy

  • 1Division of Neurosurgery, Vermont Center for Vascular Research, The University of Vermont, Burlington, VT 05405, USA.

Biochemical Pharmacology
|June 17, 2000
PubMed
Summary

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Physiological research·2016

Human white blood cells (PMNL) metabolize arachidonic acid into novel compounds. Researchers identified 20-HETE and 16(R)-HETE as key products, suggesting new pathways in cellular metabolism.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Immunology

Background:

  • Human polymorphonuclear leukocytes (PMNL) are key immune cells.
  • Arachidonic acid metabolism is crucial for cellular signaling.
  • Previous studies focused on lipoxygenase pathways in PMNL.

Purpose of the Study:

  • To identify and characterize arachidonic acid metabolites produced by intact human PMNL.
  • To investigate the enzymatic pathways involved in PMNL arachidonic acid metabolism.
  • To discover novel metabolites of arachidonic acid in PMNL.

Main Methods:

  • Incubation of intact human PMNL with arachidonic acid.
  • Isolation of metabolites using reverse-phase HPLC.
  • Structural characterization by Gas Chromatography-Mass Spectrometry (GC/MS).

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  • Stereochemical analysis using chiral-phase chromatography.

    • Main Results:

    • Four arachidonic acid metabolites were identified: 20-HETE, 16-HETE, 15-HETE, and 5-HETE.
    • 20-HETE and 16-HETE were the major metabolites, with 16-HETE being a novel finding.
    • The formation of 20-HETE was sensitive to SKF525A, indicating cytochrome P450 involvement.
    • PMNL-derived 16-HETE was identified as the 16(R)-stereoisomer.
    • 16-HETE and 20-HETE were found esterified within PMNL lipids.

    Conclusions:

    • Intact human PMNL generate 20-HETE and 16(R)-HETE via cytochrome P450 omega- and omega-4 oxygenases.
    • This contrasts with calcium ionophore-stimulated neutrophils, which primarily produce lipoxygenase products.
    • The findings reveal a novel pathway of arachidonic acid metabolism in PMNL.