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H89, an inhibitor of PKA and MSK, inhibits cyclic-AMP response element binding protein-mediated MAPK phosphatase-1 induction by lipopolysaccharide

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Abstract

Objective

Lipopolysaccharide (LPS) stimulates the production of inflammatory cytokines and the amplification of immune responses via MAPK pathways. MAPK phosphatases (MKPs) feedback-regulate the activities of MAPKs to prevent excessive immunological functions. H89 has been used as an inhibitor of the protein kinase A (PKA) and mitogen- and stress-activated protein kinase (MSK) pathways. In view of the potential roles of PKA and MSK for MKP-1 induction and the ability of H89 to inhibit these kinases, this study examined the effect of H89 on MKP-1 induction by LPS and the role of cyclic-AMP response element binding protein (CREB) in the MKP-1 induction.

Results

H89 treatment inhibited increases in MKP-1 protein and mRNA levels, and gene transcription by LPS in Raw264.7 cells. Immunoblot, gel-shift, and chromatin-immunoprecipitation assays showed the activation of CREB by LPS, and the ability of H89 to inhibit it, suggesting that H89’s inhibition of CREB may affect MKP-1 induction. In addition, H89 prevented the ability of LPS to induce other MKP genes (Dusp-2, 4, 8, and 16). Experiments using MAPK inhibitors showed that MAPKs are involved in CREB phosphorylation and MKP-1 induction, suggesting that CREB-mediated MKP-1 induction serves in part as a feedback-inhibitory loop of MAPKs.

Conclusion

Our results demonstrate that H89 inhibits the activation of CREB and the CREB-mediated MKP-1 induction by LPS, which may result from its inhibition of PKA and MSK.

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Abbreviations

CREB:

Cyclic-AMP response element binding protein

Dn:

Dominant-negative mutant

Dusp:

Dual specificity phosphatase

ERK:

Extracellular signal-regulated kinase

JNK:

c-Jun NH2-terminal kinase

LPS:

Lipopolysaccharide

MAPKs:

Mitogen-activated protein kinases

MGI:

Mouse genome informatics

MKPs:

MAPK phosphatases

MSK:

Mitogen- and stress-activated protein kinase

PKA:

Protein kinase A

TLR:

Toll-like receptor

TLRLs:

Toll-like receptor ligands

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Acknowledgements

This work was supported by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korean government (MEST) (No.R11-2007-107-01001-0), and in part by Korea Research Foundation grant KRF-2004-015-E00096, Korea.

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Authors and Affiliations

  1. College of Pharmacy, Seoul National University, Sillim-dong, Kwanak-gu, Seoul, 151-742, Korea

    Il Je Cho, Na Ri Woo & Sang Geon Kim

  2. Department of Pharmacology and Institute of Biomedical Science, College of Medicine, Hanyang University, Seoul, 133-791, Korea

    In Chul Shin

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  1. Il Je Cho
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  2. Na Ri Woo
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Correspondence to In Chul Shin or Sang Geon Kim.

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Responsible Editor: J. Di Battista.

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Cho, I.J., Woo, N.R., Shin, I.C. et al. H89, an inhibitor of PKA and MSK, inhibits cyclic-AMP response element binding protein-mediated MAPK phosphatase-1 induction by lipopolysaccharide. Inflamm. Res. 58, 863–872 (2009). https://doi.org/10.1007/s00011-009-0057-z

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  • DOI: https://doi.org/10.1007/s00011-009-0057-z

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