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Accelerative effect of a selective cyclooxygenase-2 inhibitor on Fas-mediated apoptosis in human neutrophils
Masayasu Iwase1, Sayaka Takaoka, Makiko Uchida
1Department of Oral and Maxillofacial Surgery, Showa University School of Dentistry, 2-1-1, Kitasenzoku, Ota-ku, Tokyo, 145-8515, Japan. iwase@senzoku.showa-u.ac.jp
Abstract:
Inflammatory stimuli, such as cytokines, can induce cyclooxygenase-2 (COX-2) expression in neutrophils. Selective, anti-inflammatory COX-2 inhibitors have been developed for patients with acute inflammatory diseases. Recent work has shown that selective COX-2 inhibitors interfere with tumor cell growth. The purpose of this study was to examine the capability of selective COX-2 inhibitors on Fas-mediated apoptosis in cytokine-stimulated neutrophils. Tumor necrosis factor-alpha (TNF-alpha) and granulocyte-macrophage colony-stimulating factor (GM-CSF) enhanced prostaglandin E2 (PGE2) release through the induction of COX-2 in neutrophils. This effect was not seen with either interleukin (IL)-1beta or IL-8. TNF-alpha-and GM-CSF-induced PGE2 release was blocked by the addition of the selective COX-2 inhibitor, N-(2-cyclohexyloxy-4-nitrophenyl)-methanesulfonamide (NS-398; 1 microM). GM-CSF, IL-1beta and IL-8 suppressed Fas-mediated apoptosis in neutrophils; however, this effect was not seen with TNF-alpha. The anti-apoptotic effect of cytokines on Fas-mediated neutrophil apoptosis was attenuated by the addition of NS-398 (100 microM). These results suggest that NS-398 operates via two distinct mechanisms for regulating apoptosis and COX-2 activation in neutrophils. This distinction is indicated by the difference in concentration of NS-398 required for acceleration of Fas-mediated neutrophil apoptosis, and the inhibition of PGE2 synthesis. Moreover, NS-398 suppressed the anti-apoptotic activity of IL-8 and IL-1beta, but did not induce COX-2; therefore, the pro-apoptotic mechanism of the selective COX-2 inhibitor may be unrelated to COX-2 activity. Thus, a selective COX-2 inhibitor may contribute to the reduction of acute inflammation through the enhancement of neutrophil apoptosis.
Insights
Selective COX-2 inhibitors enhance Fas-mediated apoptosis in neutrophils, potentially reducing acute inflammation. This effect may be independent of cyclooxygenase-2 (COX-2) activity, suggesting dual mechanisms of action for these anti-inflammatory drugs.
Area of Science:
- Immunology
- Pharmacology
- Cell Biology
Background:
- Cytokines like TNF-alpha and GM-CSF induce cyclooxygenase-2 (COX-2) in neutrophils.
- Selective COX-2 inhibitors are used for acute inflammatory diseases and show anti-tumor effects.
- Neutrophil apoptosis is a key process in resolving inflammation.
Purpose of the Study:
- To investigate the effect of selective COX-2 inhibitors on Fas-mediated apoptosis in cytokine-stimulated neutrophils.
- To determine if COX-2 inhibition influences cytokine-mediated suppression of neutrophil apoptosis.
- To elucidate the mechanisms by which selective COX-2 inhibitors regulate neutrophil apoptosis.
Main Methods:
- Neutrophils were stimulated with cytokines (TNF-alpha, GM-CSF, IL-1beta, IL-8).
- Prostaglandin E2 (PGE2) release was measured following cytokine stimulation.
- The selective COX-2 inhibitor NS-398 was used at different concentrations (1 microM and 100 microM).
- Fas-mediated apoptosis in neutrophils was assessed in the presence and absence of cytokines and NS-398.
Main Results:
- TNF-alpha and GM-CSF induced COX-2 and PGE2 release, which was blocked by 1 microM NS-398.
- GM-CSF, IL-1beta, and IL-8 suppressed Fas-mediated apoptosis, an effect attenuated by 100 microM NS-398.
- NS-398 at 100 microM suppressed the anti-apoptotic effects of IL-8 and IL-1beta without inducing COX-2.
Conclusions:
- Selective COX-2 inhibitors exhibit dual mechanisms in neutrophils: COX-2 inhibition at low concentrations and apoptosis modulation at higher concentrations.
- The pro-apoptotic effect of NS-398 may be independent of COX-2 inhibition, suggesting alternative pathways.
- Selective COX-2 inhibitors could reduce acute inflammation by promoting neutrophil apoptosis, offering a novel therapeutic strategy.
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