Porphyromonas gingivalis Induces Receptor Activator of NF-κB Ligand Expression in Osteoblasts through the Activator Protein 1 Pathway

Nobuo Okahashi, Hiroaki Inaba, Ichiro Nakagawa, Taihei Yamamura, Masae Kuboniwa, Koji Nakayama, Shigeyuki Hamada, Atsuo Amano

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

Porphyromonas gingivalis, an important periodontal pathogen, is closely associated with inflammatory alveolar bone resorption, and several components of the organism such as lipopolysaccharides have been reported to stimulate production of cytokines that promote inflammatory bone destruction. We investigated the effect of infection with viable P. gingivalis on cytokine production by osteoblasts. Reverse transcription-PCR and real-time PCR analyses revealed that infection with P. gingivalis induced receptor activator of nuclear factor κB (NF-κB) ligand (RANKL) mRNA expression in mouse primary osteoblasts. Production of interleukin-6 was also stimulated; however, osteoprotegerin was not. SB20350 (an inhibitor of p38 mitogen-activated protein kinase), PD98059 (an inhibitor of classic mitogen-activated protein kinase kinase, MEK1/2), wortmannin (an inhibitor of phosphatidylinositol 3 kinase), and carbobenzoxyl-leucinyl-leucinyl-leucinal (an inhibitor of NF-κB) did not prevent the RANKL expression induced by P. gingivalis. Degradation of inhibitor of NF-κB-alpha was not detectable; however, curcumin, an inhibitor of activator protein 1 (AP-1), prevented the RANKL production induced by P. gingivalis infection. Western blot analysis revealed that phosphorylation of c-Jun, a component of AP-1, occurred in the infected cells, and an analysis of c-Fos binding to an oligonucleotide containing an AP-1 consensus site also demonstrated AP-1 activation in infected osteoblasts. Infection with P. gingivalis KDP136, an isogenic deficient mutant of arginine- and lysine-specific cysteine proteinases, did not stimulate RANKL production. These results suggest that P. gingivalis infection induces RANKL expression in osteoblasts through AP-1 signaling pathways and cysteine proteases of the organism are involved in RANKL production.

Original languageEnglish
Pages (from-to)1706-1714
Number of pages9
JournalInfection and Immunity
Volume72
Issue number3
DOIs
Publication statusPublished - Mar 2004
Externally publishedYes

Fingerprint

Porphyromonas gingivalis
Transcription Factor AP-1
Cytoplasmic and Nuclear Receptors
Osteoblasts
Ligands
Infection
Cysteine Proteases
MAP Kinase Kinase 2
Phosphatidylinositol 3-Kinase
Cytokines
Alveolar Bone Loss
Osteoprotegerin
Curcumin
p38 Mitogen-Activated Protein Kinases
Protein Kinase Inhibitors
Bone Resorption
Oligonucleotides
Lysine
Reverse Transcription
Lipopolysaccharides

ASJC Scopus subject areas

  • Immunology

Cite this

Porphyromonas gingivalis Induces Receptor Activator of NF-κB Ligand Expression in Osteoblasts through the Activator Protein 1 Pathway. / Okahashi, Nobuo; Inaba, Hiroaki; Nakagawa, Ichiro; Yamamura, Taihei; Kuboniwa, Masae; Nakayama, Koji; Hamada, Shigeyuki; Amano, Atsuo.

In: Infection and Immunity, Vol. 72, No. 3, 03.2004, p. 1706-1714.

Research output: Contribution to journalArticle

Okahashi, Nobuo ; Inaba, Hiroaki ; Nakagawa, Ichiro ; Yamamura, Taihei ; Kuboniwa, Masae ; Nakayama, Koji ; Hamada, Shigeyuki ; Amano, Atsuo. / Porphyromonas gingivalis Induces Receptor Activator of NF-κB Ligand Expression in Osteoblasts through the Activator Protein 1 Pathway. In: Infection and Immunity. 2004 ; Vol. 72, No. 3. pp. 1706-1714.
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AU - Okahashi, Nobuo

AU - Inaba, Hiroaki

AU - Nakagawa, Ichiro

AU - Yamamura, Taihei

AU - Kuboniwa, Masae

AU - Nakayama, Koji

AU - Hamada, Shigeyuki

AU - Amano, Atsuo

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AB - Porphyromonas gingivalis, an important periodontal pathogen, is closely associated with inflammatory alveolar bone resorption, and several components of the organism such as lipopolysaccharides have been reported to stimulate production of cytokines that promote inflammatory bone destruction. We investigated the effect of infection with viable P. gingivalis on cytokine production by osteoblasts. Reverse transcription-PCR and real-time PCR analyses revealed that infection with P. gingivalis induced receptor activator of nuclear factor κB (NF-κB) ligand (RANKL) mRNA expression in mouse primary osteoblasts. Production of interleukin-6 was also stimulated; however, osteoprotegerin was not. SB20350 (an inhibitor of p38 mitogen-activated protein kinase), PD98059 (an inhibitor of classic mitogen-activated protein kinase kinase, MEK1/2), wortmannin (an inhibitor of phosphatidylinositol 3 kinase), and carbobenzoxyl-leucinyl-leucinyl-leucinal (an inhibitor of NF-κB) did not prevent the RANKL expression induced by P. gingivalis. Degradation of inhibitor of NF-κB-alpha was not detectable; however, curcumin, an inhibitor of activator protein 1 (AP-1), prevented the RANKL production induced by P. gingivalis infection. Western blot analysis revealed that phosphorylation of c-Jun, a component of AP-1, occurred in the infected cells, and an analysis of c-Fos binding to an oligonucleotide containing an AP-1 consensus site also demonstrated AP-1 activation in infected osteoblasts. Infection with P. gingivalis KDP136, an isogenic deficient mutant of arginine- and lysine-specific cysteine proteinases, did not stimulate RANKL production. These results suggest that P. gingivalis infection induces RANKL expression in osteoblasts through AP-1 signaling pathways and cysteine proteases of the organism are involved in RANKL production.

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