Pyrimidinoceptor potentiation of macrophage PGE 2 release involved in the induction of nitric oxide synthase

Bing C. Chen, Wan W. Lin

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

1. We have previously demonstrated that Ca 2+/calmodulin-dependent protein kinase (CaMK) mediates pyrimidinoceptor potentiation of LPS-elicited inducible nitric oxide synthase (iNOS) induction in murine J774 macrophages. In the present paper, we have explored the role of cyclooxygenase (COX)-dependent prostaglandin E 2 (PGE 2) formation in this event. 2. In J774 macrophages predominantly expressing P2Y 6 receptors, the simultaneous addition of UTP and lipopolysaccharide (LPS) resulted in potentiated increase in PGE 2 release. 3. UTP-induced increased PGE 2 release was demonstrated by a concomitant increase in COX-2 protein expression, and was decreased by inhibitors specific for phosphatidylinositide-phospholipase C (PI-PLC), CaMK, protein kinase C (PKC), nuclear factor-kappa B (NF-κB) or COX-2. 4. NS-398 (a selective COX-2 inhibitor) reduced LPS plus UTP-elicited iNOS induction and nitrite accumulation, supporting for the positive regulation of iNOS gene expression by endogenous PGE 2. 5. Moreover, the cyclic AMP/PKA-dependent up-regulation of iNOS expression mediated by PGE 2 was drawn from the inhibitory effects of 2',5'-dideoxyadenosine, KT5720 and H-89. Exogenous PGE 2 induced NF-κB activation and potentiated nitrite accumulation in response to LPS. 6. In addition to COX-2 induction, arachidonic acid (AA) release and steady-state mRNA levels of type V secretory phospholipase A 2 (sPLA 2) and Ca 2+-independent PLA 2 (iPLA 2) were also increased in the presence of LPS and UTP; the LPS-induced increase in iPLA 2 activity was also potentiated by UTP. 7. Taken together, we conclude that UTP-mediated COX-2 and iPLA 2 potentiation and PGE 2 formation contribute to the iNOS induction, and that CaMK activation is the primary step in the UTP enhancement of COX-2 induction.

Original languageEnglish
Pages (from-to)777-786
Number of pages10
JournalBritish Journal of Pharmacology
Volume130
Issue number4
Publication statusPublished - 2000
Externally publishedYes

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Uridine Triphosphate
Prostaglandins E
Nitric Oxide Synthase
Nitric Oxide Synthase Type II
Lipopolysaccharides
Cyclooxygenase 2
Macrophages
Calcium-Calmodulin-Dependent Protein Kinases
NF-kappa B
Nitrites
Phospholipases A
Cyclooxygenase 2 Inhibitors
Type C Phospholipases
Prostaglandin-Endoperoxide Synthases
Arachidonic Acid
Cyclic AMP
Protein Kinase C
Up-Regulation
Gene Expression
Messenger RNA

Keywords

  • CaMK
  • COX-2
  • iNOS
  • Macrophages
  • NF-κB
  • PGE
  • PLA
  • Pyrimidinoceptor

ASJC Scopus subject areas

  • Pharmacology

Cite this

Pyrimidinoceptor potentiation of macrophage PGE 2 release involved in the induction of nitric oxide synthase. / Chen, Bing C.; Lin, Wan W.

In: British Journal of Pharmacology, Vol. 130, No. 4, 2000, p. 777-786.

Research output: Contribution to journalArticle

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abstract = "1. We have previously demonstrated that Ca 2+/calmodulin-dependent protein kinase (CaMK) mediates pyrimidinoceptor potentiation of LPS-elicited inducible nitric oxide synthase (iNOS) induction in murine J774 macrophages. In the present paper, we have explored the role of cyclooxygenase (COX)-dependent prostaglandin E 2 (PGE 2) formation in this event. 2. In J774 macrophages predominantly expressing P2Y 6 receptors, the simultaneous addition of UTP and lipopolysaccharide (LPS) resulted in potentiated increase in PGE 2 release. 3. UTP-induced increased PGE 2 release was demonstrated by a concomitant increase in COX-2 protein expression, and was decreased by inhibitors specific for phosphatidylinositide-phospholipase C (PI-PLC), CaMK, protein kinase C (PKC), nuclear factor-kappa B (NF-κB) or COX-2. 4. NS-398 (a selective COX-2 inhibitor) reduced LPS plus UTP-elicited iNOS induction and nitrite accumulation, supporting for the positive regulation of iNOS gene expression by endogenous PGE 2. 5. Moreover, the cyclic AMP/PKA-dependent up-regulation of iNOS expression mediated by PGE 2 was drawn from the inhibitory effects of 2',5'-dideoxyadenosine, KT5720 and H-89. Exogenous PGE 2 induced NF-κB activation and potentiated nitrite accumulation in response to LPS. 6. In addition to COX-2 induction, arachidonic acid (AA) release and steady-state mRNA levels of type V secretory phospholipase A 2 (sPLA 2) and Ca 2+-independent PLA 2 (iPLA 2) were also increased in the presence of LPS and UTP; the LPS-induced increase in iPLA 2 activity was also potentiated by UTP. 7. Taken together, we conclude that UTP-mediated COX-2 and iPLA 2 potentiation and PGE 2 formation contribute to the iNOS induction, and that CaMK activation is the primary step in the UTP enhancement of COX-2 induction.",
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