Dependence and Reversal of Nitric Oxide Production on NFKB in Silica and Lipopolysaccharide-Induced Macrophages

"Summary: In this report the differential regulation of NF-KB and nitric oxide (NO) was investigated in the mouse macrophage cell line RAW 264.7 following exposure to a mineral dust (silica) and /or an endotoxin (Lipopolysaccharide, LPS). The results indicated that silica and LPS can significantly induce the activation of NF-KB as well as elicit enhanced production of NO in RAW 264.7 cells as part of an early inflammatory response mechanism. A 24-hour time-course study showed that NO release from these cells continued to increase following the initial stimulus by LPS or silica. In contrast, activation of NF-KB was maximal at 6 hours and then showed a steady decline to 24 hours. The production of NO was suppressed by protease inhibitor and antioxidant, both of which block the activation of NF-KB. Surprisingly, the use of an NO synthase inhibitor resulted in an enhancement of NF-JCB activation. These findings suggest that NO produced in macrophage cells in response to an inflammatory stimulus like silica or LPS my be linked to a negative feedback role on the activation of NF-KB. It is now accepted that nitric oxide (NO) plays a pivotal role in the inflammation response process when macrophage cells are activated by a pro-inflammatory stimulus (l). Production of NO is dependent on the activity of the enzyme nitric oxide synthase (NOS). Up until now, at least three distinct genes have been described which encode different isoforms of NOS. NOS has been cloned from a variety of different mammalian cells, including constitutive neuron NOS, constitutive endothelium NOS and inducible NOS (2). Macrophages appear to be the principal cellular source for the inducible form of NOS (iNOS) activated in response to an inflammatory producing event. The mechanisms subserving the regulation of the iNOS gene and NO production have been under intense investigation in recent years (3-5). Several binding sites for transcriptional factors have been identified in the promoter region of iNOS gene, including NF-lCB, AP-1, NF-IL6, Oct-I to cite a few (5). NF-KB appears to play a"