title: NF kappa b signaling in posthypoxic endothelial cells: relevance to E-selectin expression and neutrophil adhesion.
abstract: Our previous studies have implicated the nuclear transcription factor kappa B (NF kappa B) in the regulation of adhesion molecule expression in endothelial cells exposed to anoxia-reoxygenation (A/R) or a redox imbalance. The objectives of this study were (1) to define the kinetics of NF kappa B activation by examining I kappa B alpha degradation and the nuclear translocation of p65 in response to A/R or redox imbalance (induced by treatment of cells with diamide and buthionine sulfoximine) and (2) to determine whether the signal for I kappa B alpha degradation, nuclear translocation of p65, and E-selectin-mediated neutrophil adhesion is related to the activity of protein tyrosine kinase (PTK), protein tyrosine phosphatase (PTPase) and/or protein kinase C (PKC). The results demonstrate that both A/R and redox imbalance led to I kappa B alpha degradation within 30 min and the concomitant appearance of p65 in the nucleus, consistent with rapid cytosolic activation of NF kappa B and subsequent nuclear translocation of the activated p65 subunit. Inhibition of PKC blocked I kappa B alpha degradation and p65 translocation in A/R-challenged, but not redox-altered, endothelial cells. However, both A/R- and redox-induced NF kappa B activation was blocked by inhibition of PTK. Similarly, A/R-induced E-selectin expression and neutrophil-endothelial cell adhesion were blocked by inhibition of PKC or PTK, while only PTK inhibited the redox-induced adhesion response. Pretreatment of cells with N-acetyl cysteine effectively blocked A/R- or redox-induced I kappa B degradation and significantly attenuated the respective neutrophil adhesion responses. Collectively, these findings indicate that A/R-induced E-selectin expression and neutrophil-endothelial cell adhesion are mediated by both PKC and PTK, which signal rapid activation of NF kappa B. This A/R-induced NF kappa B signaling response appears to be mediated, at least in part, by intracellular redox imbalance.
authors.affiliation: Department of Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA.
authors.forename: S
authors.initials: S
authors.lastname: Kokura
authors.forename: C A
authors.initials: CA
authors.lastname: Rhoads
authors.forename: R E
authors.initials: RE
authors.lastname: Wolf
authors.forename: T
authors.initials: T
authors.lastname: Yoshikawa
authors.forename: D N
authors.initials: DN
authors.lastname: Granger
authors.forename: T Y
authors.initials: TY
authors.lastname: Aw
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002448:Cell Adhesion; D015687:Cell Hypoxia; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte; D004268:DNA-Binding Proteins / Q000378:metabolism; D003958:Diamide / Q000494:pharmacology; D019040:E-Selectin / Q000096:biosynthesis* / Q000235:genetics; D004730:Endothelium, Vascular / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D020672:I-kappa B Proteins; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism* / Q000502:physiology*; D009504:Neutrophils / Q000166:cytology*; D010084:Oxidation-Reduction; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology; D051996:Transcription Factor RelA; D014471:Umbilical Veins
chemical_list: D004268:DNA-Binding Proteins; D019040:E-Selectin; D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D051996:Transcription Factor RelA; D003958:Diamide; D000072000:NF-KappaB Inhibitor alpha; D019328:Buthionine Sulfoximine; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C; D017027:Protein Tyrosine Phosphatases; D000111:Acetylcysteine