Triclosan inhibits tumor necrosis factor-alpha-stimulated urokinase production in human gingival fibroblasts.
OBJECTIVE
Destruction of the supporting periodontal tissues is mediated by the action of several proteolytic enzymes. Urokinase is a serine protease that plays a key role in connective tissue destruction through conversion of plasminogen into plasmin. The present study was conducted to evaluate the effect of triclosan on the production and activity of urokinase in cultured gingival fibroblasts.


METHODS
Urokinase production was studied in primary cultures of human gingival fibroblasts stimulated with tumor necrosis factor-alpha. Urokinase activity and production were evaluated using casein zymography and western blotting, respectively. Urokinase mRNA expression was evaluated using the reverse transcription-polymerase chain reaction. Triclosan was used to interfere with this stimulatory effect. The roles of different cell-signaling cascades involved in urokinase production were assessed through western blotting and immunofluorescence using several cell-signaling inhibitors.


RESULTS
Tumor necrosis factor-alpha was found to be a strong stimulus for urokinase production and triclosan was able to inhibit this response at the protein and mRNA levels. Triclosan was also able to inhibit conversion of plasminogen into plasmin. Tumor necrosis factor-alpha-stimulated urokinase production was shown to be dependent on the nuclear factor-kappaB and c-Jun N-terminal kinase signaling pathways. Triclosan inhibited c-Jun N-terminal kinase phosphorylation and c-Jun production.


CONCLUSIONS
Within the limits of this study, these results show that triclosan may inhibit urokinase production and plasminogen activation in gingival fibroblasts through modulation of the c-Jun N-terminal kinase signaling pathway.
Arancibia
R
R
Laboratory of Periodontal Physiology, Dentistry Academic Unit, School of Medicine, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
Cáceres
M
M
Martínez
J
J
Smith
P C
PC
D000873:Anthracenes / Q000494:pharmacology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D005341:Fibrinolysin / Q000037:antagonists & inhibitors; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology; D005419:Flavonoids / Q000494:pharmacology; D005881:Gingiva / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D048369:MAP Kinase Kinase 1 / Q000037:antagonists & inhibitors; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D010455:Peptides / Q000494:pharmacology; D010766:Phosphorylation; D010958:Plasminogen / Q000037:antagonists & inhibitors; D012333:RNA, Messenger / Q000032:analysis / Q000187:drug effects; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D014260:Triclosan / Q000008:administration & dosage / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors*
D000873:Anthracenes; D004791:Enzyme Inhibitors; D005419:Flavonoids; D016328:NF-kappa B; D010455:Peptides; D012333:RNA, Messenger; C118258:SN50 peptide; D015842:Serine Proteinase Inhibitors; D014409:Tumor Necrosis Factor-alpha; C432165:pyrazolanthrone; D014260:Triclosan; D010958:Plasminogen; D048031:JNK Mitogen-Activated Protein Kinases; D048369:MAP Kinase Kinase 1; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one