Immune Netw.  2013 Dec;13(6):283-288. 10.4110/in.2013.13.6.283.

BIRB 796 has Distinctive Anti-inflammatory Effects on Different Cell Types

• Ryoo S
• Choi J
• Kim J
• Bae S
• Hong J
• Jo S
• Kim S
• Lee Y

• KMID: 2150789
• DOI: http://doi.org/10.4110/in.2013.13.6.283

Abstract

The pro-inflammatory cytokines tumor necrosis factor-alpha (TNFalpha) and interleukin (IL)-1beta are crucial mediators involved in chronic inflammatory diseases. Inflammatory signal pathways regulate inflammatory cytokine expression-mediated by p38 mitogen activated protein kinase (p38MAPK). Therefore, considerable attention has been given to p38MAPK as a target molecule for the development of a novel anti-inflammatory therapeutics. BIRB 796, one of p38MAPK inhibitor, is a candidate of therapeutic drug for chronic inflammatory diseases. In this study, we investigated the effect of BIRB 796 on inflammatory cytokine productions by lipopolysaccharide (LPS) in different immune cell types. BIRB 796 reduced LPS-mediated IL-8 production in THP-1 cells but not in Raw 264.7 cells. Further analysis of signal molecules by western blot revealed that BIRB 796 sufficiently suppressed LPS-mediated phosphorylation of p38MAPK in both cell types whereas it failed to block inhibitor of kappa B (I-kappaB) degradation in Raw 264.7 cells. Taken together, these results suggest that the anti-inflammatory function of BIRB 796 depends on cell types.

Keyword

p38MAPK; BIRB 796; Inflammatory cytokine; Inflammatory signal pathways; LPS

MeSH Terms

Blotting, Western
Cytokines
Interleukin-8
Interleukins
Phosphorylation
Protein Kinases
Signal Transduction
Tumor Necrosis Factor-alpha
Cytokines
Interleukin-8
Interleukins
Protein Kinases
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1 Inhibitory effect of BIRB 796 and AGI 1067 on cytokine production in Raw 264.7 and THP-1 cells. Raw 264.7 and THP-1 cells were pre-incubated with BIRB 796 and AGI 1067 (10µM) for 1 h prior to stimulation of LPS (10 ng/ml) for Raw 264.7 and LPS (1µg/ml) for THP-1 cells. Levels of mouse TNFα (A), mouse IL-1β (B) and human IL-8 (C) were determined by ELISA kits from R&D Systems. Mean±SEM of cytokine levels were measured (**p<0.01; ***p <0.001). Representative data from 1 of 3 independent experiments performed in duplicate are shown.

• 

  Figure 2 Inhibitory effect of BIRB 796 and AGI 1067 on cytokine transcripts in Raw 264.7 and THP-1 cells. Raw 264.7 and THP-1 cells were pre-incubated with BIRB 796 and AGI 1067 (10µM) for 1 h and then stimulated with LPS (10 ng/ml) for Raw 264.7 and LPS (1µg/ml) for THP-1 cells for 4 hrs. The transcript of cytokines in Raw 264.7 (A) and THP-1 (B) were detected by RT-PCR. Representative data from 1 of 3 independent experiments are shown.

• 

  Figure 3 Effect of BIRB 796 and AGI 1067 on the activation of p38MAPK and degradation of I-κB in Raw 264.7 and THP-1 cells. (A) Raw 264.7 and (B) THP-1 cells were pre-incubated with BIRB 796 and AGI 1067 (10µM) for 1 h and then stimulated with LPS (10 ng/ml) for Raw 264.7 and LPS (1µg/ml) for THP-1 cells for 30 min. The cells were lyzed and total cell lysates of 20µg per lane were electrophoresed in 10% SDS-PAGE. The phosphorylation of p38MAPK and degradation of I-κB were detected using western blot with specific antibodies. Representative data from 1 of 3 independent experiments are shown.

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