S-nitrosoglutathione inhibits TNF- alpha -induced NF Kappa B activation in neutrophils

• Published in: Inflammation research 2001-02, Vol.50 (2), p.89-95
• Main Authors: tenberry, J D, Owens, M L, Chen, N X, Brown, LAS
• Format: Article
• Language: English
• Subjects: NF-^KB protein; S-nitrosoglutathione
• ISSN: 1023-3830; 1420-908X
• DOI: 10.1007/s000110050729

Objective and Design: Cytokine expression is controlled by transcription factors including NF Kappa B, which has recently been found to exist in human neutrophils. We previously showed that exogenous nitric oxide (NO) induces neutrophil apoptosis and hypothesized that this NO effect could be mediated by inhibition of NF Kappa B activation.Materials and methods: Isolated human neutrophils were incubated with or without S-nitrosoglutathione (GSNO 0.1mM-5mM; Sigma) for 2 h. Neutrophils were either unstimulated or stimulated with TNF alpha or n-formyl methionyl leucine phenylalanine (fMLP). Viability was assessed by vital dye cytotoxicity assay. After nuclear extraction and measurement of protein concentration, NF Kappa B binding was determined by electrophoretic mobility shift assay. Effects of GSNO on activation of I Kappa B alpha , which inhibits intranuclear translocation of NF Kappa B, were measured by Western immunoblot technique. For comparison, experiments were also performed in the presence of the NF Kappa B inhibitor PDTC.Results: TNF alpha increased nuclear NF Kappa B activity compared to unstimulated neutrophils (p