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B and AP-1 Activation That Promotes Cyclooxygenase-2 Expression1
Department of Medicine IV, Experimental Division, Faculty of Medicine, University of Erlangen-Nürnberg, Erlangen, Germany
Macrophages are a major source of cytokines and proinflammatory
radicals such as superoxide. These mediators can be both produced and
utilized by macrophages in autocrine-regulatory pathways. Therefore, we
studied the potential role of oxygen radical-regulatory mechanisms in
reprogramming macrophage apoptosis. Preactivation of RAW 264.7 cells
with a nontoxic dose of the redox cycler
2,3-dimethoxy-1,4-naphtoquinone (5 µM) for 15 h attenuated
S-nitrosoglutathione (1 mM)-initiated apoptotic cell
death and averted accumulation of the tumor suppressor p53, which is
indicative for macrophage apoptosis. Preactivation with superoxide
promoted cyclooxygenase-2 induction that was NF-
B and AP-1 mediated.
NF-B activation was confirmed by p50/p65-heterodimer formation,
IB-
degradation, and stimulation of a NF-B luciferase reporter
construct. Furthermore, a NF-B decoy approach abrogated
cyclooxygenase-2 (Cox-2) expression as well as inducible protection.
The importance of AP-1 for superoxide-mediated Cox-2 expression and
cell protection was substantiated by using the extracellular
signal-regulated kinase-inhibitor PD98059 and the p38-inhibitor
SB203580, which blocked Cox-2 expression. In corroboration, Cox-2
expression was hindered by a dominant-negative c-jun
mutant (TAM67). Protection from apoptosis was verified in human
macrophages with the notion that superoxide promoted Cox-2 expression,
which in turn attenuated nitric oxide-evoked caspase activation. We
conclude that the sublethal generation of oxygen radicals reprograms
macrophages by NF-B and AP-1 activation. The resulting
hyporesponsiveness reveals an attenuated apoptotic program in
association with Cox-2 expression.
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