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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

TNF-α release from RAW 264 cells after LPS stimulation in the presence or absence of serum and its inhibition by GGA. Cells were incubated with LPS (1 µg/ml) in the presence (closed column) or absence (open column) of serum for 4 h at 37°C under a 5% CO2 and 95% air atmosphere. GGA (80 µM) was added 2 h before the addition of LPS. TNF-α level was measured as described in Materials and Methods. Data points represent the means ± SE (n = 3). ap<0.01 vs LPS alone-treated cells in the presence of serum. bp<0.05 vs LPS alone-treated cells in the absence of serum.
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Figure 9: TNF-α release from RAW 264 cells after LPS stimulation in the presence or absence of serum and its inhibition by GGA. Cells were incubated with LPS (1 µg/ml) in the presence (closed column) or absence (open column) of serum for 4 h at 37°C under a 5% CO2 and 95% air atmosphere. GGA (80 µM) was added 2 h before the addition of LPS. TNF-α level was measured as described in Materials and Methods. Data points represent the means ± SE (n = 3). ap<0.01 vs LPS alone-treated cells in the presence of serum. bp<0.05 vs LPS alone-treated cells in the absence of serum.

Mentions: Serum proteins such as soluble LBP and soluble CD14 play important roles in cell activation induced by LPS. To investigate the involvement of serum proteins in the inhibitory action of GGA, TNF-α production by RAW 264 cells was determined in the presence or absence of serum (Fig. 9). TNF-α release at 4 h after LPS (1 µg/ml) challenge under serum-free condition reduced to one third of that in the presence of serum. However, GGA significantly inhibited TNF-α production regardless of the presence of serum. Furthermore, GGA failed to suppress the binding of LPS to LBP on in vitro study although CL as a positive control strongly inhibited LPS-LBP binding (data not shown).


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)

TNF-α release from RAW 264 cells after LPS stimulation in the presence or absence of serum and its inhibition by GGA. Cells were incubated with LPS (1 µg/ml) in the presence (closed column) or absence (open column) of serum for 4 h at 37°C under a 5% CO2 and 95% air atmosphere. GGA (80 µM) was added 2 h before the addition of LPS. TNF-α level was measured as described in Materials and Methods. Data points represent the means ± SE (n = 3). ap<0.01 vs LPS alone-treated cells in the presence of serum. bp<0.05 vs LPS alone-treated cells in the absence of serum.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 9: TNF-α release from RAW 264 cells after LPS stimulation in the presence or absence of serum and its inhibition by GGA. Cells were incubated with LPS (1 µg/ml) in the presence (closed column) or absence (open column) of serum for 4 h at 37°C under a 5% CO2 and 95% air atmosphere. GGA (80 µM) was added 2 h before the addition of LPS. TNF-α level was measured as described in Materials and Methods. Data points represent the means ± SE (n = 3). ap<0.01 vs LPS alone-treated cells in the presence of serum. bp<0.05 vs LPS alone-treated cells in the absence of serum.
Mentions: Serum proteins such as soluble LBP and soluble CD14 play important roles in cell activation induced by LPS. To investigate the involvement of serum proteins in the inhibitory action of GGA, TNF-α production by RAW 264 cells was determined in the presence or absence of serum (Fig. 9). TNF-α release at 4 h after LPS (1 µg/ml) challenge under serum-free condition reduced to one third of that in the presence of serum. However, GGA significantly inhibited TNF-α production regardless of the presence of serum. Furthermore, GGA failed to suppress the binding of LPS to LBP on in vitro study although CL as a positive control strongly inhibited LPS-LBP binding (data not shown).

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