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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

Upregulation of TSP-1 in THP-1 cells by P. gingivalis LPS stimulation.(A) THP-1 cells were stimulated by P. gingivalis LPS at concentrations of 0, 0.0001, 0.001, 0.01, 0.1, or 1.0 µg/ml for 4 h. P. gingivalis LPS increased TSP-1 mRNA expression in a dose-dependent manner in THP-1 cells. (B) THP-1 cells were stimulated by 1.0 µg/ml of P. gingivalis LPS for 0, 1, 2, 4, 12, 24, 48, or 72 h. P. gingivalis LPS increased TSP-1 mRNA expression in a time-dependent manner in THP-1 cells. (C) THP-1 cells were stimulated with P. gingivalis LPS at concentrations of 0, 0.01, 0.1, or 1.0 µg/ml for 72 h. P. gingivalis LPS increased TSP-1 protein production in a dose-dependent manner in THP-1 cells. (D) THP-1 cells were stimulated with 1.0 µg/ml of P. gingivalis LPS for 0, 1, 4, 6, 12, 24, 48, or 72 h. P. gingivalis LPS increased TSP-1 protein production in a time-dependent manner in THP-1 cells. (E) PMA-treated THP-1 cells were stimulated with P. gingivalis LPS at concentrations of 0, 0.01, 0.1, or 1.0 µg/ml for 72 h. P. gingivalis LPS increased TSP-1 protein production in a dose-dependent manner in PMA-treated THP-1 cells.
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pone-0115107-g002: Upregulation of TSP-1 in THP-1 cells by P. gingivalis LPS stimulation.(A) THP-1 cells were stimulated by P. gingivalis LPS at concentrations of 0, 0.0001, 0.001, 0.01, 0.1, or 1.0 µg/ml for 4 h. P. gingivalis LPS increased TSP-1 mRNA expression in a dose-dependent manner in THP-1 cells. (B) THP-1 cells were stimulated by 1.0 µg/ml of P. gingivalis LPS for 0, 1, 2, 4, 12, 24, 48, or 72 h. P. gingivalis LPS increased TSP-1 mRNA expression in a time-dependent manner in THP-1 cells. (C) THP-1 cells were stimulated with P. gingivalis LPS at concentrations of 0, 0.01, 0.1, or 1.0 µg/ml for 72 h. P. gingivalis LPS increased TSP-1 protein production in a dose-dependent manner in THP-1 cells. (D) THP-1 cells were stimulated with 1.0 µg/ml of P. gingivalis LPS for 0, 1, 4, 6, 12, 24, 48, or 72 h. P. gingivalis LPS increased TSP-1 protein production in a time-dependent manner in THP-1 cells. (E) PMA-treated THP-1 cells were stimulated with P. gingivalis LPS at concentrations of 0, 0.01, 0.1, or 1.0 µg/ml for 72 h. P. gingivalis LPS increased TSP-1 protein production in a dose-dependent manner in PMA-treated THP-1 cells.

Mentions: Fig. 2 demonstrates that TSP-1 expression was enhanced by P. gingivalis LPS in THP-1 cells. The highest expression was observed at a concentration of 1.0 µg/ml (Fig. 2A). TSP-1 mRNA expression at concentrations of 1.0 µg/ml and 0.1 µg/ml P. gingivalis LPS was significantly different from that observed without stimulation (P<0.05 and P<0.001, respectively). The highest expression was observed at 4 h of stimulation with the P. gingivalis LPS preparation (Fig. 2B). THP-1 cells with P. gingivalis LPS preparations reached a peak after 4 h; this corresponds to the fact that TSP-1 is transiently released early during the acute inflammatory phase. The secretion of TSP-1 protein was increased in a time and dose-dependent manner (Fig. 2C, 2D). TSP-1 mRNA expression was also upregulated in PMA-treated THP-1 cells stimulated with P. gingivalis LPS (Fig. 2E).


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)

Upregulation of TSP-1 in THP-1 cells by P. gingivalis LPS stimulation.(A) THP-1 cells were stimulated by P. gingivalis LPS at concentrations of 0, 0.0001, 0.001, 0.01, 0.1, or 1.0 µg/ml for 4 h. P. gingivalis LPS increased TSP-1 mRNA expression in a dose-dependent manner in THP-1 cells. (B) THP-1 cells were stimulated by 1.0 µg/ml of P. gingivalis LPS for 0, 1, 2, 4, 12, 24, 48, or 72 h. P. gingivalis LPS increased TSP-1 mRNA expression in a time-dependent manner in THP-1 cells. (C) THP-1 cells were stimulated with P. gingivalis LPS at concentrations of 0, 0.01, 0.1, or 1.0 µg/ml for 72 h. P. gingivalis LPS increased TSP-1 protein production in a dose-dependent manner in THP-1 cells. (D) THP-1 cells were stimulated with 1.0 µg/ml of P. gingivalis LPS for 0, 1, 4, 6, 12, 24, 48, or 72 h. P. gingivalis LPS increased TSP-1 protein production in a time-dependent manner in THP-1 cells. (E) PMA-treated THP-1 cells were stimulated with P. gingivalis LPS at concentrations of 0, 0.01, 0.1, or 1.0 µg/ml for 72 h. P. gingivalis LPS increased TSP-1 protein production in a dose-dependent manner in PMA-treated THP-1 cells.
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Related In: Results  -  Collection

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

pone-0115107-g002: Upregulation of TSP-1 in THP-1 cells by P. gingivalis LPS stimulation.(A) THP-1 cells were stimulated by P. gingivalis LPS at concentrations of 0, 0.0001, 0.001, 0.01, 0.1, or 1.0 µg/ml for 4 h. P. gingivalis LPS increased TSP-1 mRNA expression in a dose-dependent manner in THP-1 cells. (B) THP-1 cells were stimulated by 1.0 µg/ml of P. gingivalis LPS for 0, 1, 2, 4, 12, 24, 48, or 72 h. P. gingivalis LPS increased TSP-1 mRNA expression in a time-dependent manner in THP-1 cells. (C) THP-1 cells were stimulated with P. gingivalis LPS at concentrations of 0, 0.01, 0.1, or 1.0 µg/ml for 72 h. P. gingivalis LPS increased TSP-1 protein production in a dose-dependent manner in THP-1 cells. (D) THP-1 cells were stimulated with 1.0 µg/ml of P. gingivalis LPS for 0, 1, 4, 6, 12, 24, 48, or 72 h. P. gingivalis LPS increased TSP-1 protein production in a time-dependent manner in THP-1 cells. (E) PMA-treated THP-1 cells were stimulated with P. gingivalis LPS at concentrations of 0, 0.01, 0.1, or 1.0 µg/ml for 72 h. P. gingivalis LPS increased TSP-1 protein production in a dose-dependent manner in PMA-treated THP-1 cells.
Mentions: Fig. 2 demonstrates that TSP-1 expression was enhanced by P. gingivalis LPS in THP-1 cells. The highest expression was observed at a concentration of 1.0 µg/ml (Fig. 2A). TSP-1 mRNA expression at concentrations of 1.0 µg/ml and 0.1 µg/ml P. gingivalis LPS was significantly different from that observed without stimulation (P<0.05 and P<0.001, respectively). The highest expression was observed at 4 h of stimulation with the P. gingivalis LPS preparation (Fig. 2B). THP-1 cells with P. gingivalis LPS preparations reached a peak after 4 h; this corresponds to the fact that TSP-1 is transiently released early during the acute inflammatory phase. The secretion of TSP-1 protein was increased in a time and dose-dependent manner (Fig. 2C, 2D). TSP-1 mRNA expression was also upregulated in PMA-treated THP-1 cells stimulated with P. gingivalis LPS (Fig. 2E).

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