Modulation of Macrophage Nitric Oxide Production by Prostaglandin D2

Charles F. Bellows, Adam Alder, Peter Wludyka, Bernard M. Jaffe

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


Background: Nitric oxide and prostaglandins readily become activated in response to inflammatory events. The overproduction of nitric oxide is detrimental to the host. The present study was conducted to examine whether prostaglandin D2 (PGD2) modulates nitric oxide production in macrophages in response to an inflammatory stimulus. Methods: Cultures of RAW 264.7 murine macrophages were exposed to Escherichia coli lipopolysaccharide (LPS, 0.01 and 1.0 μg/ml) before and after exposure to PGD2 (0.01 to 10 nmol). After 24-h incubation, supernatants were collected and nitrite was quantitated by Greiss reaction as a measure of nitric oxide synthesis. Inducible nitric oxide synthase (iNOS) protein was measured by Western blot analysis. Results: Macrophages exposed to 0.01 and 1.0 μg/ml LPS produced 8.3 ± 0.2 and 15.0 ± 1.4 nmol/1.1 × 106 cells/24 h of nitrite, respectively. The simultaneous addition of PGD2 with LPS inhibited nitrite production in a dose-dependent fashion and suppressed iNOS protein expression. A strong time effect was also exhibited when macrophages were incubated with PGD2 1 hour before as compared to 7 hours after the addition of LPS (0.01 or 1.0 μg/ml), indicating that the earlier the time PGD2 was added to the culture media, the greater the inhibition. Prostaglandin D2 had the capacity to block nitrite synthesis even when added as much as 7 hours after an LPS challenge. Blocking endogenous prostaglandins, using indomethacin (10 μΜ), suppressed nitrite production. Conclusion: Exogenous PGD2 caused dose- and time-dependent decreases in LPS-stimulated nitrite production by RAW 264.7 macrophages by hindering iNOS protein expression. Conversely, the endogenous prostaglandins released by these same cells in response to an LPS challenge stimulated nitrite production, which may consequently dampen the inhibitory actions of exogenous PGD2.

Original languageEnglish (US)
Pages (from-to)92-97
Number of pages6
JournalJournal of Surgical Research
Issue number1
StatePublished - May 2006


  • macrophages
  • nitric oxide
  • prostaglandin D
  • sepsis

ASJC Scopus subject areas

  • Surgery


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