We have examined the effects of lipopolysaccharide (LPS) from Prevotella nigrescens, one of the causative agents of inflammatory periodontal disease and endodontic infections, on the production of nitric oxide (NO) and expression of inducible nitric oxide synthase (iNOS) in the murine macrophage cell line RAW264.7. We also attempted to throw light on the signaling mechanisms involved in P. nigrescens LPS-induced NO production. We found that P. nigrescens LPS can induce iNOS expression and stimulate the release of NO without additional stimuli and demonstrated an important role of the transcription factor NF-kappaB and microtubule polymerization in NO production. The production of NO required l-arginine and protein tyrosine kinase but not activation of protein kinase C. The ability of P. nigrescens LPS to promote the production of NO may be important in the pathogenesis of inflammatory periodontal disease and endodontic infections.