Enhancement of phosphorylation and transcriptional activity of the glucocorticoid receptor in human synovial fibroblasts by nimesulide, a preferential cyclooxygenase 2 inhibitor
Ja. Di Battista et al., Enhancement of phosphorylation and transcriptional activity of the glucocorticoid receptor in human synovial fibroblasts by nimesulide, a preferential cyclooxygenase 2 inhibitor, ARTH RHEUM, 42(1), 1999, pp. 157-166
Objective. To examine the effect of 2 nonsteroidal antiinflammatory drugs (
NSAIDs), nimesulide (NIM), a preferential cyclooxygenase 2 (COX-2) inhibito
r, and naproxen (NAP), on the functional parameters and transcriptional act
ivity of the glucocorticoid receptor (GR) system in cultured human synovial
fibroblasts (HSF).
Methods. HSF were incubated with NIR;I (0.3, 3, and 30 mu g/ml), NAP (15, 3
0, and 90 mu g/ml), and dexamethasone (DEX; 0.01, 0.1, and 1 mu M) on a tim
e- and dose-dependent basis. The numbers of GR binding sites per cell were
determined by radioligand receptor assay. Total cellular, cytoplasmic, or n
uclear GR protein was measured by Western analysis using a specific anti-hu
man GR antibody. Phosphorylation of GR was determined by specific immunopre
cipitation of protein extracts from P-32-orthophosphate-labeled HSF. Mitoge
n-activated protein kinase p44/42 (MAPK) phosphorylation was followed by We
stern analysis using a specific anti-phosphoMAPK antibody. Levels of activa
ted nuclear GR capable of binding specifically to a P-32-labeled oligonucle
otide harboring the glucocorticoid/hormone response element (GRE) were eval
uated by gel electrophoretic mobility shift analysis. The effects of NIM an
d DEX on transcriptional activation of the mouse mammary tumor virus (MMTV)
promoter was determined by transfecting HSF with MMTV-luciferase (reporter
gene) constructs.
Results. NIM had no effect on the number of GR binding sites, in contrast t
o NAP and DEX. NIM and NAP did not influence cellular GR protein levels or
nucleocytoplasmic shuttling, although DEX lowered GR messenger RNA and prot
ein levels after 48 hours. NIR;I, but not NAP, markedly increased MAPK phos
phorylation (suggesting an increase in MAPK cascade activity), GR phosphory
lation, GR binding to GRE, and transcriptional activation of MMTV promoter
through the GRE site in the promoter,
Conclusion. This study is the first to report that the antiinflammatory eff
ects of NLM, an NSAID, may be partly related to its activation of the GR sy
stem.