CYCLOOXYGENASE 2 (COX-2) GENE ACTIVATION IS REGULATED BY CYCLIC ADENOSINE MONOPHOSPHATE

Publisher Website
Google Scholar
Abstract
Prostaglandin E2 production by tissue-fixed macrophages (MØ) after severe injury contributes to an enhanced susceptibility to infection and sepsis. The purpose of this study was to investigate the effect of cyclic adenosine monophosphate (cAMP) on prostaglandin (PGE2) production and cyclooxygenase II (COX-2) gene activation in LPS-stimulated macrophages (MØ). RAW264.7 cells, a mouse MØ cell line, were exposed to various concentrations of dibutyryl cAMP ± lipopolysaccharide (10 μg/mL) stimulation. Total MØ ribonucleic acid (RNA) was harvested for the determination of COX-2 messenger RNA (mRNA) with mouse complementary deoxyribonucleic acid (cDNA) by Northern blot assay. MØ supernatant was collected for the measurement of tumor necrosis factor (TNF) by L929 bioassay and PGE2 by enzyme-linked immunosorbent assay (ELISA), respectively. MØ NFκB activity was determined by electrophoretic mobility shift assays (EMSA). Dibutyryl cAMP significantly inhibited TNF production by LPS-stimulated MØ. Dibutyryl cAMP (1 mM) alone induced PGE2 production. Dibutyryl cAMP (100 μM and 1 mM) also augmented PGE2 production by LPS-stimulated MØ. Dibutyryl cAMP had similar effect on MØ COX-2 mRNA expression and NFκB activity. Our data demonstrate that cAMP modulates MØ TNF production and upregulates COX-2 gene and PGE2 production.

This publication has 0 references indexed in Scilit: