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Related Experiment Videos

L-ascorbic acid modulates 5-lipoxygenase activity in human polymorphonuclear leukocytes.

K H Schmidt1, D Steinhilber, U Moser

  • 1Department of Surgery, University of Tübingen, FRG.

International Archives of Allergy and Applied Immunology
|January 1, 1988
PubMed
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Vitamin C (L-ascorbate) inhibits inflammatory mediators like Leukotriene B4 (LTB4) by affecting 5-lipoxygenase activity in neutrophils. It also influences LTB4 production during phagocytosis.

Area of Science:

  • Biochemistry
  • Immunology
  • Cell Biology

Background:

  • Leukotriene B4 (LTB4) is a key inflammatory mediator derived from arachidonic acid via 5-lipoxygenase.
  • Polymorphonuclear neutrophils (PMNL) release LTB4 upon stimulation.
  • LTB4 plays a significant role in mediating inflammatory responses.

Purpose of the Study:

  • To investigate the effect of extracellular L-ascorbate (Vitamin C) on LTB4 and 5-HETE biosynthesis in PMNL.
  • To explore how L-ascorbate influences LTB4 production under different stimulation conditions (ionophore and phagocytosis).
  • To characterize the transport mechanism of L-ascorbate into PMNL.

Main Methods:

  • Measurement of LTB4 and 5-HETE production in PMNL stimulated with calcium ionophore A 23187 and opsonized zymosan.

Related Experiment Videos

  • Assessment of 5-lipoxygenase activity and LTB4 catabolism.
  • Kinetic analysis of L-ascorbate uptake by PMNL, including stereospecificity.

    • Main Results:

    • Extracellular L-ascorbate (0.5-5.0 mM) inhibits ionophore-stimulated LTB4 and 5-HETE production.
    • L-ascorbate reverses the deactivation of 5-lipoxygenase caused by zymosan, increasing LTB4 and 5-HETE yields.
    • L-ascorbate transport into PMNL is stereospecific, saturable (Km 39 microM, Vmax 0.28 nmol/10(8) cells.min), and includes passive diffusion (P = 1.75 microliter/10(8) cells.min).

    Conclusions:

    • Extracellular L-ascorbate modulates inflammatory mediator production by PMNL, impacting 5-lipoxygenase activity.
    • The concentration-dependent effects of L-ascorbate suggest a dual role in regulating LTB4 biosynthesis.
    • Understanding L-ascorbate uptake kinetics is crucial for its intracellular functions and potential therapeutic applications in inflammation.