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Geranylgeranylacetone ameliorates inflammatory response to lipopolysaccharide (LPS) in murine macrophages: inhibition of LPS binding to the cell surface.

Mochida S, Matsura T, Yamashita A, Horie S, Ohata S, Kusumoto C, Nishida T, Minami Y, Inagaki Y, Ishibe Y, Nakada J, Ohta Y, Yamada K - J Clin Biochem Nutr (2007)

Bottom Line: GGA (80 microM) treatment 2 h before LPS addition significantly suppressed TNF-alpha and NO productions at 12 h and 24 h after LPS, respectively, indicating that GGA inhibits activation of macrophages.When it was examined whether GGA interacts with LPS and/or affects expression of Toll-like receptor 4 (TLR4) and CD14 on the cell surface, GGA inhibited the binding of LPS to the cell surface, while GGA did not affect TLR4 and CD14 expressions.These results indicate that GGA suppresses the binding of LPS to the cell surface of macrophages, resulting in inhibiting signal transduction downstream of TLR4.

View Article: PubMed Central - PubMed

Affiliation: Division of Medical Biochemistry, Department of Pathophysiological and Therapeutic Science, Tottori University Faculty of Medicine, 86 Nishi-cho, Yonago 683-8503, Japan.

ABSTRACT
We investigated whether pretreatment with geranylgeranylacetone (GGA), a potent heat shock protein (HSP) inducer, could inhibit proinflammatory cytokine liberation and nitric oxide (NO) production in lipopolysaccharide (LPS)-treated murine macrophages. The levels of NO and tumor necrosis factor-alpha (TNF-alpha) released from murine macrophage RAW 264 cells were increased dose- and time-dependently following treatment with LPS (1 microg/ml). GGA (80 microM) treatment 2 h before LPS addition significantly suppressed TNF-alpha and NO productions at 12 h and 24 h after LPS, respectively, indicating that GGA inhibits activation of macrophages. However, replacement by fresh culture medium before LPS treatment abolished the inhibitory effect of GGA on NO production in LPS-treated cells. Furthermore, GGA inhibited both HSP70 and inducible NO synthase expressions induced by LPS treatment despite an HSP inducer. When it was examined whether GGA interacts with LPS and/or affects expression of Toll-like receptor 4 (TLR4) and CD14 on the cell surface, GGA inhibited the binding of LPS to the cell surface, while GGA did not affect TLR4 and CD14 expressions. These results indicate that GGA suppresses the binding of LPS to the cell surface of macrophages, resulting in inhibiting signal transduction downstream of TLR4.

No MeSH data available.


Related in: MedlinePlus

Effects of GGA on binding of LPS to the cell surface in RAW 264 cells. Cells were incubated with 80 µM GGA for 2 h at 37°C and for further 30 min after addition of LPS (1 µg/ml). Cells were washed, and stained with anti-LPS antibody, followed by goat anti-mouse IgG-FITC. The LPS fluorescence resulting from LPS binding to cells was measured by using Epics cytofluorometer. Representative data from three separate experiments are shown.
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Figure 7: Effects of GGA on binding of LPS to the cell surface in RAW 264 cells. Cells were incubated with 80 µM GGA for 2 h at 37°C and for further 30 min after addition of LPS (1 µg/ml). Cells were washed, and stained with anti-LPS antibody, followed by goat anti-mouse IgG-FITC. The LPS fluorescence resulting from LPS binding to cells was measured by using Epics cytofluorometer. Representative data from three separate experiments are shown.

Mentions: To investigate the effect of GGA on LPS binding to the cell surface, we used flow cytometry with an anti-LPS monoclonal antibody (Fig. 7). Cell surface LPS increased after LPS stimulation. However, the LPS binding was inhibited when LPS was added to the medium in the presence of GGA.


Geranylgeranylacetone ameliorates inflammatory response to lipopolysaccharide (LPS) in murine macrophages: inhibition of LPS binding to the cell surface.

Mochida S, Matsura T, Yamashita A, Horie S, Ohata S, Kusumoto C, Nishida T, Minami Y, Inagaki Y, Ishibe Y, Nakada J, Ohta Y, Yamada K - J Clin Biochem Nutr (2007)

Effects of GGA on binding of LPS to the cell surface in RAW 264 cells. Cells were incubated with 80 µM GGA for 2 h at 37°C and for further 30 min after addition of LPS (1 µg/ml). Cells were washed, and stained with anti-LPS antibody, followed by goat anti-mouse IgG-FITC. The LPS fluorescence resulting from LPS binding to cells was measured by using Epics cytofluorometer. Representative data from three separate experiments are shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2170953&req=5

Figure 7: Effects of GGA on binding of LPS to the cell surface in RAW 264 cells. Cells were incubated with 80 µM GGA for 2 h at 37°C and for further 30 min after addition of LPS (1 µg/ml). Cells were washed, and stained with anti-LPS antibody, followed by goat anti-mouse IgG-FITC. The LPS fluorescence resulting from LPS binding to cells was measured by using Epics cytofluorometer. Representative data from three separate experiments are shown.
Mentions: To investigate the effect of GGA on LPS binding to the cell surface, we used flow cytometry with an anti-LPS monoclonal antibody (Fig. 7). Cell surface LPS increased after LPS stimulation. However, the LPS binding was inhibited when LPS was added to the medium in the presence of GGA.

Bottom Line: GGA (80 microM) treatment 2 h before LPS addition significantly suppressed TNF-alpha and NO productions at 12 h and 24 h after LPS, respectively, indicating that GGA inhibits activation of macrophages.When it was examined whether GGA interacts with LPS and/or affects expression of Toll-like receptor 4 (TLR4) and CD14 on the cell surface, GGA inhibited the binding of LPS to the cell surface, while GGA did not affect TLR4 and CD14 expressions.These results indicate that GGA suppresses the binding of LPS to the cell surface of macrophages, resulting in inhibiting signal transduction downstream of TLR4.

View Article: PubMed Central - PubMed

Affiliation: Division of Medical Biochemistry, Department of Pathophysiological and Therapeutic Science, Tottori University Faculty of Medicine, 86 Nishi-cho, Yonago 683-8503, Japan.

ABSTRACT
We investigated whether pretreatment with geranylgeranylacetone (GGA), a potent heat shock protein (HSP) inducer, could inhibit proinflammatory cytokine liberation and nitric oxide (NO) production in lipopolysaccharide (LPS)-treated murine macrophages. The levels of NO and tumor necrosis factor-alpha (TNF-alpha) released from murine macrophage RAW 264 cells were increased dose- and time-dependently following treatment with LPS (1 microg/ml). GGA (80 microM) treatment 2 h before LPS addition significantly suppressed TNF-alpha and NO productions at 12 h and 24 h after LPS, respectively, indicating that GGA inhibits activation of macrophages. However, replacement by fresh culture medium before LPS treatment abolished the inhibitory effect of GGA on NO production in LPS-treated cells. Furthermore, GGA inhibited both HSP70 and inducible NO synthase expressions induced by LPS treatment despite an HSP inducer. When it was examined whether GGA interacts with LPS and/or affects expression of Toll-like receptor 4 (TLR4) and CD14 on the cell surface, GGA inhibited the binding of LPS to the cell surface, while GGA did not affect TLR4 and CD14 expressions. These results indicate that GGA suppresses the binding of LPS to the cell surface of macrophages, resulting in inhibiting signal transduction downstream of TLR4.

No MeSH data available.


Related in: MedlinePlus