Limits...
A novel chromone derivative with anti-inflammatory property via inhibition of ROS-dependent activation of TRAF6-ASK1-p38 pathway.

Liu H, Xu R, Feng L, Guo W, Cao N, Qian C, Teng P, Wang L, Wu X, Sun Y, Li J, Shen Y, Xu Q - PLoS ONE (2012)

Bottom Line: In the present study, we show that a novel chromone derivative, DCO-6, significantly reduced lipopolysaccharide (LPS)-induced production of nitric oxide, IL-1β and IL-6, decreased the levels of iNOS, IL-1β and IL-6 mRNA expression in both RAW264.7 cells and mouse primary peritoneal macrophages, and inhibited LPS-induced activation of p38 MAPK but not of JNK, ERK.LPS-induced production of intracellular reactive oxygen species (ROS) was remarkably impaired by DCO-6, which disrupted the formation of the TRAF6-ASK1 complex.Our results indicate that DCO-6 showed anti-inflammatory properties through inhibition of ROS-dependent activation of TRAF6-ASK1-p38 pathway.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, China.

ABSTRACT
The p38 MAPK signaling pathway plays a pivotal role in inflammation. Targeting p38 MAPK may be a potential strategy for the treatment of inflammatory diseases. In the present study, we show that a novel chromone derivative, DCO-6, significantly reduced lipopolysaccharide (LPS)-induced production of nitric oxide, IL-1β and IL-6, decreased the levels of iNOS, IL-1β and IL-6 mRNA expression in both RAW264.7 cells and mouse primary peritoneal macrophages, and inhibited LPS-induced activation of p38 MAPK but not of JNK, ERK. Moreover, DCO-6 specifically inhibited TLR4-dependent p38 activation without directly inhibiting its kinase activity. LPS-induced production of intracellular reactive oxygen species (ROS) was remarkably impaired by DCO-6, which disrupted the formation of the TRAF6-ASK1 complex. Administering DCO-6 significantly protected mice from LPS-induced septic shock in parallel with the inhibition of p38 activation and ROS production. Our results indicate that DCO-6 showed anti-inflammatory properties through inhibition of ROS-dependent activation of TRAF6-ASK1-p38 pathway. Blockade of the upstream events required for p38 MAPK action by DCO-6 may provide a new therapeutic option in the treatment of inflammatory diseases.

Show MeSH

Related in: MedlinePlus

Inhibition of LPS-induced NO, IL-1β and IL-6 production by DCO-6 in murine macrophages.RAW264.7 cells or peritoneal macrophages from BALB/c mice were treated with various concentrations of DCO-6 in the absence or presence of LPS. (A) The levels of NO, IL-1β and IL-6 in the cell culture medium were determined 24 h after LPS stimulation as described in Methods. (B) The levels of iNOS, IL-1β and IL-6 mRNA were determined by real-time quantitative PCR 8 h after LPS stimulation. β-actin was used as an invariant control. Data are shown as means ± S.D. of three independent experiments. *P<0.05 vs LPS control.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3376149&req=5

pone-0037168-g002: Inhibition of LPS-induced NO, IL-1β and IL-6 production by DCO-6 in murine macrophages.RAW264.7 cells or peritoneal macrophages from BALB/c mice were treated with various concentrations of DCO-6 in the absence or presence of LPS. (A) The levels of NO, IL-1β and IL-6 in the cell culture medium were determined 24 h after LPS stimulation as described in Methods. (B) The levels of iNOS, IL-1β and IL-6 mRNA were determined by real-time quantitative PCR 8 h after LPS stimulation. β-actin was used as an invariant control. Data are shown as means ± S.D. of three independent experiments. *P<0.05 vs LPS control.

Mentions: To assess the effects of DCO-6 on LPS-induced production of cellular mediator in RAW264.7 and peritoneal macrophages, cell culture medium was harvested. Measuring nitrite as the index of NO production by the Griess method, we found that LPS treatment for 24 h resulted in a large amount of NO release in macrophages (Fig. 2A). Co-incubation of DCO-6 with LPS inhibited the formation of NO in a concentration-dependent manner. In addition, LPS-induced production of IL-1β and IL-6 was significantly reduced in macrophages treated with DCO-6 (Fig. 2A). When RNA was isolated and quantitative real-time PCR was performed to examine the effects of DCO-6 on gene expression, Co-incubation of DCO-6 with LPS also decreased the levels of iNOS, IL-1β and IL-6 mRNA expression (Fig. 2B), suggesting that NO, IL-1β and IL-6 reduction by DCO-6 may be related to transcriptional inhibition. However, neither TNF-α release nor mRNA induction was altered by DCO-6 at concentration of up to 30 µM (Fig. S3).


A novel chromone derivative with anti-inflammatory property via inhibition of ROS-dependent activation of TRAF6-ASK1-p38 pathway.

Liu H, Xu R, Feng L, Guo W, Cao N, Qian C, Teng P, Wang L, Wu X, Sun Y, Li J, Shen Y, Xu Q - PLoS ONE (2012)

Inhibition of LPS-induced NO, IL-1β and IL-6 production by DCO-6 in murine macrophages.RAW264.7 cells or peritoneal macrophages from BALB/c mice were treated with various concentrations of DCO-6 in the absence or presence of LPS. (A) The levels of NO, IL-1β and IL-6 in the cell culture medium were determined 24 h after LPS stimulation as described in Methods. (B) The levels of iNOS, IL-1β and IL-6 mRNA were determined by real-time quantitative PCR 8 h after LPS stimulation. β-actin was used as an invariant control. Data are shown as means ± S.D. of three independent experiments. *P<0.05 vs LPS control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3376149&req=5

pone-0037168-g002: Inhibition of LPS-induced NO, IL-1β and IL-6 production by DCO-6 in murine macrophages.RAW264.7 cells or peritoneal macrophages from BALB/c mice were treated with various concentrations of DCO-6 in the absence or presence of LPS. (A) The levels of NO, IL-1β and IL-6 in the cell culture medium were determined 24 h after LPS stimulation as described in Methods. (B) The levels of iNOS, IL-1β and IL-6 mRNA were determined by real-time quantitative PCR 8 h after LPS stimulation. β-actin was used as an invariant control. Data are shown as means ± S.D. of three independent experiments. *P<0.05 vs LPS control.
Mentions: To assess the effects of DCO-6 on LPS-induced production of cellular mediator in RAW264.7 and peritoneal macrophages, cell culture medium was harvested. Measuring nitrite as the index of NO production by the Griess method, we found that LPS treatment for 24 h resulted in a large amount of NO release in macrophages (Fig. 2A). Co-incubation of DCO-6 with LPS inhibited the formation of NO in a concentration-dependent manner. In addition, LPS-induced production of IL-1β and IL-6 was significantly reduced in macrophages treated with DCO-6 (Fig. 2A). When RNA was isolated and quantitative real-time PCR was performed to examine the effects of DCO-6 on gene expression, Co-incubation of DCO-6 with LPS also decreased the levels of iNOS, IL-1β and IL-6 mRNA expression (Fig. 2B), suggesting that NO, IL-1β and IL-6 reduction by DCO-6 may be related to transcriptional inhibition. However, neither TNF-α release nor mRNA induction was altered by DCO-6 at concentration of up to 30 µM (Fig. S3).

Bottom Line: In the present study, we show that a novel chromone derivative, DCO-6, significantly reduced lipopolysaccharide (LPS)-induced production of nitric oxide, IL-1β and IL-6, decreased the levels of iNOS, IL-1β and IL-6 mRNA expression in both RAW264.7 cells and mouse primary peritoneal macrophages, and inhibited LPS-induced activation of p38 MAPK but not of JNK, ERK.LPS-induced production of intracellular reactive oxygen species (ROS) was remarkably impaired by DCO-6, which disrupted the formation of the TRAF6-ASK1 complex.Our results indicate that DCO-6 showed anti-inflammatory properties through inhibition of ROS-dependent activation of TRAF6-ASK1-p38 pathway.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, China.

ABSTRACT
The p38 MAPK signaling pathway plays a pivotal role in inflammation. Targeting p38 MAPK may be a potential strategy for the treatment of inflammatory diseases. In the present study, we show that a novel chromone derivative, DCO-6, significantly reduced lipopolysaccharide (LPS)-induced production of nitric oxide, IL-1β and IL-6, decreased the levels of iNOS, IL-1β and IL-6 mRNA expression in both RAW264.7 cells and mouse primary peritoneal macrophages, and inhibited LPS-induced activation of p38 MAPK but not of JNK, ERK. Moreover, DCO-6 specifically inhibited TLR4-dependent p38 activation without directly inhibiting its kinase activity. LPS-induced production of intracellular reactive oxygen species (ROS) was remarkably impaired by DCO-6, which disrupted the formation of the TRAF6-ASK1 complex. Administering DCO-6 significantly protected mice from LPS-induced septic shock in parallel with the inhibition of p38 activation and ROS production. Our results indicate that DCO-6 showed anti-inflammatory properties through inhibition of ROS-dependent activation of TRAF6-ASK1-p38 pathway. Blockade of the upstream events required for p38 MAPK action by DCO-6 may provide a new therapeutic option in the treatment of inflammatory diseases.

Show MeSH
Related in: MedlinePlus