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Thrombospondin-1 production is enhanced by Porphyromonas gingivalis lipopolysaccharide in THP-1 cells.

Gokyu M, Kobayashi H, Nanbara H, Sudo T, Ikeda Y, Suda T, Izumi Y - PLoS ONE (2014)

Bottom Line: Recent studies have suggested that TSP-1 is associated with rheumatoid arthritis, diabetes mellitus, and osteoclastogenesis.Furthermore, IL-17F synergistically enhanced P. gingivalis LPS-induced TSP-1 production.It may also contribute a new target molecule for periodontal therapy.

View Article: PubMed Central - PubMed

Affiliation: Periodontology, Bio-Matrix Department, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.

ABSTRACT
Periodontitis is a chronic inflammatory disease caused by gram-negative anaerobic bacteria. Monocytes and macrophages stimulated by periodontopathic bacteria induce inflammatory mediators that cause tooth-supporting structure destruction and alveolar bone resorption. In this study, using a DNA microarray, we identified the enhanced gene expression of thrombospondin-1 (TSP-1) in human monocytic cells stimulated by Porphyromonas gingivalis lipopolysaccharide (LPS). TSP-1 is a multifunctional extracellular matrix protein that is upregulated during the inflammatory process. Recent studies have suggested that TSP-1 is associated with rheumatoid arthritis, diabetes mellitus, and osteoclastogenesis. TSP-1 is secreted from neutrophils, monocytes, and macrophages, which mediate immune responses at inflammatory regions. However, TSP-1 expression in periodontitis and the mechanisms underlying TSP-1 expression in human monocytic cells remain unknown. Here using real-time RT-PCR, we demonstrated that TSP-1 mRNA expression level was significantly upregulated in inflamed periodontitis gingival tissues and in P. gingivalis LPS-stimulated human monocytic cell line THP-1 cells. TSP-1 was expressed via Toll-like receptor (TLR) 2 and TLR4 pathways. In P. gingivalis LPS stimulation, TSP-1 expression was dependent upon TLR2 through the activation of NF-κB signaling. Furthermore, IL-17F synergistically enhanced P. gingivalis LPS-induced TSP-1 production. These results suggest that modulation of TSP-1 expression by P. gingivalis plays an important role in the progression and chronicity of periodontitis. It may also contribute a new target molecule for periodontal therapy.

No MeSH data available.


Related in: MedlinePlus

P. gingivalis LPS-induced TSP-1 mRNA expression was inhibited by TLR2-neutralizing antibody in THP-1 cells.THP-1 cells were pretreated with 10 ng/ml of IgG2a isotype control, TLR2-neutralizing antibody, and TLR4-neutralizing antibody for 30 min, followed by the addition of 1.0 µg/ml of P. gingivalis LPS, Pam2CSK4, and E. coli LPS for 4 h. (A) TLR2-neutralizing antibody and TLR4-neutralizing antibody neutralized the activity of Pam2CSK4 and E. coli LPS, respectively. (B) P. gingivalis LPS with TLR2-neutralizing antibody significantly reduced TSP-1 mRNA expression compared with P. gingivalis LPS alone in THP-1 cells (P<0.001).
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pone-0115107-g004: P. gingivalis LPS-induced TSP-1 mRNA expression was inhibited by TLR2-neutralizing antibody in THP-1 cells.THP-1 cells were pretreated with 10 ng/ml of IgG2a isotype control, TLR2-neutralizing antibody, and TLR4-neutralizing antibody for 30 min, followed by the addition of 1.0 µg/ml of P. gingivalis LPS, Pam2CSK4, and E. coli LPS for 4 h. (A) TLR2-neutralizing antibody and TLR4-neutralizing antibody neutralized the activity of Pam2CSK4 and E. coli LPS, respectively. (B) P. gingivalis LPS with TLR2-neutralizing antibody significantly reduced TSP-1 mRNA expression compared with P. gingivalis LPS alone in THP-1 cells (P<0.001).

Mentions: To examine whether P. gingivalis LPS required TLR2 or TLR4, THP-1 cells were stimulated with 1.0 µg/ml of P. gingivalis LPS, Pam2CSK4 (TLR2 ligand), or E. coli LPS (TLR4 ligand) for 4 h. All 3 stimulatory factors significantly increased TSP-1 mRNA expression (P<0.001; Fig. 3). This result suggested that P. gingivalis LPS utilized both TLR2 and TLR4 pathways in this study. Furthermore, THP-1 cells were pretreated with 10 ng/ml of isotype control, anti-TLR2, and anti-TLR4 for 30 min, followed by treatment with 1.0 µg/ml of P. gingivalis LPS, Pam2CSK4, and E. coli LPS for 4 h. Anti-TLR2 and anti-TLR4 significantly inhibited TSP-1 mRNA expression by Pam2CSK4 and E. coli LPS, respectively (P<0.001; Fig. 4A). Anti-TLR2 significantly reduced P. gingivalis LPS-induced TSP-1 mRNA expression (P<0.001; Fig. 4B). Anti-TLR4 had little influence on expression (Fig. 4B). These data indicated that P. gingivalis LPS-induced TSP-1 expression was upregulated in THP-1 cells via the TLR2 pathway.


Thrombospondin-1 production is enhanced by Porphyromonas gingivalis lipopolysaccharide in THP-1 cells.

Gokyu M, Kobayashi H, Nanbara H, Sudo T, Ikeda Y, Suda T, Izumi Y - PLoS ONE (2014)

P. gingivalis LPS-induced TSP-1 mRNA expression was inhibited by TLR2-neutralizing antibody in THP-1 cells.THP-1 cells were pretreated with 10 ng/ml of IgG2a isotype control, TLR2-neutralizing antibody, and TLR4-neutralizing antibody for 30 min, followed by the addition of 1.0 µg/ml of P. gingivalis LPS, Pam2CSK4, and E. coli LPS for 4 h. (A) TLR2-neutralizing antibody and TLR4-neutralizing antibody neutralized the activity of Pam2CSK4 and E. coli LPS, respectively. (B) P. gingivalis LPS with TLR2-neutralizing antibody significantly reduced TSP-1 mRNA expression compared with P. gingivalis LPS alone in THP-1 cells (P<0.001).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4264871&req=5

pone-0115107-g004: P. gingivalis LPS-induced TSP-1 mRNA expression was inhibited by TLR2-neutralizing antibody in THP-1 cells.THP-1 cells were pretreated with 10 ng/ml of IgG2a isotype control, TLR2-neutralizing antibody, and TLR4-neutralizing antibody for 30 min, followed by the addition of 1.0 µg/ml of P. gingivalis LPS, Pam2CSK4, and E. coli LPS for 4 h. (A) TLR2-neutralizing antibody and TLR4-neutralizing antibody neutralized the activity of Pam2CSK4 and E. coli LPS, respectively. (B) P. gingivalis LPS with TLR2-neutralizing antibody significantly reduced TSP-1 mRNA expression compared with P. gingivalis LPS alone in THP-1 cells (P<0.001).
Mentions: To examine whether P. gingivalis LPS required TLR2 or TLR4, THP-1 cells were stimulated with 1.0 µg/ml of P. gingivalis LPS, Pam2CSK4 (TLR2 ligand), or E. coli LPS (TLR4 ligand) for 4 h. All 3 stimulatory factors significantly increased TSP-1 mRNA expression (P<0.001; Fig. 3). This result suggested that P. gingivalis LPS utilized both TLR2 and TLR4 pathways in this study. Furthermore, THP-1 cells were pretreated with 10 ng/ml of isotype control, anti-TLR2, and anti-TLR4 for 30 min, followed by treatment with 1.0 µg/ml of P. gingivalis LPS, Pam2CSK4, and E. coli LPS for 4 h. Anti-TLR2 and anti-TLR4 significantly inhibited TSP-1 mRNA expression by Pam2CSK4 and E. coli LPS, respectively (P<0.001; Fig. 4A). Anti-TLR2 significantly reduced P. gingivalis LPS-induced TSP-1 mRNA expression (P<0.001; Fig. 4B). Anti-TLR4 had little influence on expression (Fig. 4B). These data indicated that P. gingivalis LPS-induced TSP-1 expression was upregulated in THP-1 cells via the TLR2 pathway.

Bottom Line: Recent studies have suggested that TSP-1 is associated with rheumatoid arthritis, diabetes mellitus, and osteoclastogenesis.Furthermore, IL-17F synergistically enhanced P. gingivalis LPS-induced TSP-1 production.It may also contribute a new target molecule for periodontal therapy.

View Article: PubMed Central - PubMed

Affiliation: Periodontology, Bio-Matrix Department, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.

ABSTRACT
Periodontitis is a chronic inflammatory disease caused by gram-negative anaerobic bacteria. Monocytes and macrophages stimulated by periodontopathic bacteria induce inflammatory mediators that cause tooth-supporting structure destruction and alveolar bone resorption. In this study, using a DNA microarray, we identified the enhanced gene expression of thrombospondin-1 (TSP-1) in human monocytic cells stimulated by Porphyromonas gingivalis lipopolysaccharide (LPS). TSP-1 is a multifunctional extracellular matrix protein that is upregulated during the inflammatory process. Recent studies have suggested that TSP-1 is associated with rheumatoid arthritis, diabetes mellitus, and osteoclastogenesis. TSP-1 is secreted from neutrophils, monocytes, and macrophages, which mediate immune responses at inflammatory regions. However, TSP-1 expression in periodontitis and the mechanisms underlying TSP-1 expression in human monocytic cells remain unknown. Here using real-time RT-PCR, we demonstrated that TSP-1 mRNA expression level was significantly upregulated in inflamed periodontitis gingival tissues and in P. gingivalis LPS-stimulated human monocytic cell line THP-1 cells. TSP-1 was expressed via Toll-like receptor (TLR) 2 and TLR4 pathways. In P. gingivalis LPS stimulation, TSP-1 expression was dependent upon TLR2 through the activation of NF-κB signaling. Furthermore, IL-17F synergistically enhanced P. gingivalis LPS-induced TSP-1 production. These results suggest that modulation of TSP-1 expression by P. gingivalis plays an important role in the progression and chronicity of periodontitis. It may also contribute a new target molecule for periodontal therapy.

No MeSH data available.


Related in: MedlinePlus