The reduction of Fusobacterium nucleatum in mice is irrelevant to the nitric oxide induced by iNOS.
Previously we reported that mice infected recurrently with live Fusobacterim nucleatum (Fn) synthesize a significant amount of NO between 12 hr and 24 hr after the Fn injection. We now investigated whether the NO has the capability of killing Fn, a gram-negative rod periodontal pathogen. The mice were divided into three groups: treated with live bacteria (LB), treated with heat-killed bacteria (HKB) and untreated: normal (N). The Fn reduction, NO production and cell number after Fn injection were then compared in these mice. In the LB group, no Fn was detected at 6 hr, whereas it was still detected in the HKB and N groups at 24 hr as assessed by both colony counts and PCR assays. A significant amount of NO was synthesized in the LB group at 24 hr after the Fn injection. Fn is not killed by SNAP-generated NO in vitro. An increase in the total cell number was accompanied by an increase of the neutrophil numbers in the LB group. Intracellular O2(-) generation (including ONOO(-)) was visualized using dihydrorhodamine (DHR)-123. The peak of O2(-) generation by PEC was shown to be at 3 hr in all 3 groups. The number of O2(-) positive cells in the LB group at 3 hr was remarkably high, and most of them were likely to be neutrophils. The Fn reduction would be performed cooperatively via oxygen dependent and oxygen independent mechanisms. Thus reactive oxygen species (ROS) included in the oxygen dependent mechanism appear to be important for Fn reduction. However the significant amounts of NO derived from the iNOS synthesized in the LB group between 12 hr and 24 hr after injection of LFn were not involved in the Fn reduction.
Kato
Chihomi
C
Department of Microbiology, School of Dentistry, Nippon Dental University at Niigata, Niigata, Niigata 951-8580, Japan. ckato@ngt.ndu.ac.jp
Mikami
Masato
M
Suzuki
Anri
A
Saito
Kazuko
K
D000818:Animals; D002452:Cell Count; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005674:Fusobacterium Infections / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism; D016967:Fusobacterium nucleatum / Q000254:growth & development*; D016131:Lymphocyte Subsets / Q000276:immunology / Q000382:microbiology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009569:Nitric Oxide / Q000096:biosynthesis / Q000276:immunology* / Q000494:pharmacology; D020030:Nitric Oxide Donors / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D010529:Peritoneal Cavity / Q000382:microbiology; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012235:Rhodamines / Q000378:metabolism; D026423:S-Nitroso-N-Acetylpenicillamine / Q000494:pharmacology; D013047:Specific Pathogen-Free Organisms; D013481:Superoxides / Q000378:metabolism
D004269:DNA, Bacterial; D020030:Nitric Oxide Donors; D012336:RNA, Ribosomal, 16S; D012235:Rhodamines; C058319:dihydrorhodamine 123; D013481:Superoxides; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse