Co-stimulatory effect of nitric oxide on endothelial NF-kappaB implies a physiological self-amplifying mechanism.

Here we investigated the effects of the second messenger molecule NO at various concentrations on the activation of transcription factor NF-kappaB, IkappaB-alpha kinase (IKK-alpha), Jun N-terminal kinase (JNK) and apoptosis in murine endothelial cells. Low concentrations of NO alone failed to activate NF-kappaB, IKK-alpha and JNK. When NF-kappaB was prestimulated by TNF-alpha or phorbol 12-myristate 13-acetate, the addition of NO at low concentrations enhanced the activation of NF-kappaB. This provides a mechanism for a self-amplifying signal in the inflammatory response, since the inducible NO synthase in endothelial cells is regulated by NF-kappaB. The co-stimulatory effect of NO on NF-kappaB activation was also evident from IKK-alpha kinase assays and reporter gene experiments in endothelial cells. High doses of NO impaired the TNF-alpha-induced DNA-binding activity of NF-kappaB. Accordingly, these high amounts of NO also repressed the TNF-alpha-induced transactivation by NF-kappaB as efficient as dexamethasone. The doses of NO required for the inhibition of NF-kappaB are not cytotoxic for the endothelial cells, enabling the establishment of an autoregulatory loop for NF-kappaB signaling.