title: Extracellular β-NAD(+) inhibits interleukin-1-induced matrix metalloproteinase-1 and -3 expression on human gingival fibroblasts.
abstract: There is increasing evidence to show that extracellular β-nicotinamide adenine dinucleotide (β-NAD(+)) modulates various biological functions in inflammatory/immune regions. The aim of this study was to determine the effect of β-NAD(+) on matrix metalloproteinase (MMP) expression on human gingival fibroblasts (hGF), the excess production of which leads to the matrix degradation associated with the pathological processes of periodontitis. The expression of MMP-1 and MMP-3 on hGF was determined by real-time polymerase chain reaction (PCR) and an enzyme-linked immunosorbent assay. The phosphorylated status of mitogen-activated protein (MAP) kinases, extracellular signal-regulated kinase 1/2 (ERK), c-Jun N-terminal kinase (JNK), and p38 and the expression of inhibitor κB (IκB)α were determined by Western blotting. β-NAD(+) inhibited the expression of MMP-1 and MMP-3 triggered by IL-1α at gene and protein levels. β-NAD(+) had no significant effect on the IL-1α-induced phosphorylation of ERK1/2, JNK, and p38 and also had no effect on the IL-1α-induced degradation of IκBα relative to the control, suggesting that inhibition by β-NAD(+) was independent of the MAP kinase and the nuclear factor-κB signaling pathways. Transcripts of NAD(+)-metabolizing enzymes, such as NAD(+)-glycohydrolase, adenosine diphosphate (ADP)-ribosylcyclase, and ADP-ribosyltransferase, were expressed by hGF as assessed by RT-PCR. Experiments using α-NAD(+), which is not a substrate for ADP-ribosylcyclase or ADP-ribosyltransferase, revealed the possible contribution of NAD(+)-glycohydrolase to the inhibition of MMP. This is consistent with the finding that ADP-ribose, an NAD(+)-metabolite by NAD(+)-glycohydrolase, exhibited MMP inhibition similar to β-NAD(+). The present findings may provide an additional viewpoint to clarify a natural feedback mechanism during the inflammatory process in periodontal tissue.
authors.affiliation: Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Gotoh
authors.forename: Eiji
authors.initials: E
authors.lastname: Nemoto
authors.forename: Sousuke
authors.initials: S
authors.lastname: Kanaya
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Shimauchi
mesh_terms: D000293:Adolescent; D000328:Adult; D005110:Extracellular Space / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005881:Gingiva / Q000166:cytology*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism*; D019278:Matrix Metalloproteinase 3 / Q000235:genetics / Q000378:metabolism*; D009243:NAD / Q000494:pharmacology*; D016328:NF-kappa B / Q000378:metabolism; D055815:Young Adult
chemical_list: D007375:Interleukin-1; D016328:NF-kappa B; D009243:NAD; C511768:MMP3 protein, human; D019278:Matrix Metalloproteinase 3; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1