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Research ArticleINFLAMMATION AND IMMUNOPHARMACOLOGY

Priming of Alveolar Macrophage Respiratory Burst by H2O2 Is Prevented by Phosphatidylcholine-Specific Phospholipase C Inhibitor Tricyclodecan-9-yl-xanthate (D609)

Julio Girón-Calle, Kousthub Srivatsa and Henry Jay Forman
Journal of Pharmacology and Experimental Therapeutics April 2002, 301 (1) 87-94; DOI: https://doi.org/10.1124/jpet.301.1.87
Julio Girón-Calle
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Kousthub Srivatsa
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Henry Jay Forman
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Abstract

The respiratory burst in alveolar macrophages is enhanced in vitro by pre-exposure to nontoxic concentrations of hydroperoxides before stimulation by an agonist, which may represent a feed-forward regulatory mechanism. Tricyclodecan-9-yl-xanthate (D609), an inhibitor of phosphatidylcholine-specific phospholipase C (PC-PLC), suppresses this priming of the respiratory burst by pre-exposure to H2O2 in NR8383 alveolar macrophages (up to 100 μM D609, 400 nmol of H2O2 added to 5 × 106 cells 15 min before stimulation with ADP). D609 has potential as an antioxidant due to its dithiocarbonate functional group that allows it to slowly react with H2O2 and rapidly reduce cytochrome c, which interferes with a common assay for the respiratory burst. Nonetheless, the antioxidant properties of D609 do not account for its inhibition of priming of the respiratory burst by H2O2. Reduction of nitro blue tetrazolium is the basis for an assay for superoxide production with which D609 does not interfere. With this assay, it was found that D609 does not inhibit the respiratory burst per se, but prevents its enhancement by pre-exposure to H2O2. Consistent with a role of diacylglycerol generation by phospholipase C, this enhancement was mimicked by pre-exposure to phorbol ester. In contrast with priming, receptor-mediated stimulation of the respiratory burst depends on the better characterized phosphatidylinositol-specific phospholipase C. Priming of the respiratory burst by H2O2 joins the list of inflammatory responses that are inhibited by D609. Nevertheless, the results herein indicate that caution should be exercised in the interpretation of the effects of D609 to consider both antioxidant effects and inhibition of PC-PLC.

Footnotes

  • This work was supported by Grant HL37556 from the National Institutes of Health.

  • Abbreviations:
    D609
    tricyclodecan-9-yl-xanthate
    PC-PLC
    phosphatidylcholine-specific phospholipase C
    [Ca2+]c
    cytosolic concentration of Ca2+
    U73122
    1-(6-((17β-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione
    PI-PLC
    phosphatidylinositol-specific phospholipase
    PBS
    phosphate-buffered saline
    BSA
    bovine serum albumin
    KRP
    Krebs-Ringer phosphate buffer
    NBT
    nitro blue tetrazolium
    PMA
    phorbol-12-myristate-13-acetate
    • Received September 7, 2001.
    • Accepted December 5, 2001.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 301 (1)
Journal of Pharmacology and Experimental Therapeutics
Vol. 301, Issue 1
1 Apr 2002
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Research ArticleINFLAMMATION AND IMMUNOPHARMACOLOGY

Priming of Alveolar Macrophage Respiratory Burst by H2O2 Is Prevented by Phosphatidylcholine-Specific Phospholipase C Inhibitor Tricyclodecan-9-yl-xanthate (D609)

Julio Girón-Calle, Kousthub Srivatsa and Henry Jay Forman
Journal of Pharmacology and Experimental Therapeutics April 1, 2002, 301 (1) 87-94; DOI: https://doi.org/10.1124/jpet.301.1.87

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Research ArticleINFLAMMATION AND IMMUNOPHARMACOLOGY

Priming of Alveolar Macrophage Respiratory Burst by H2O2 Is Prevented by Phosphatidylcholine-Specific Phospholipase C Inhibitor Tricyclodecan-9-yl-xanthate (D609)

Julio Girón-Calle, Kousthub Srivatsa and Henry Jay Forman
Journal of Pharmacology and Experimental Therapeutics April 1, 2002, 301 (1) 87-94; DOI: https://doi.org/10.1124/jpet.301.1.87
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