Exp Mol Med.  2008 Oct;40(5):574-581. 10.3858/emm.2008.40.5.574.

Histone deacetylase inhibitor KBH-A42 inhibits cytokine production in RAW 264.7 macrophage cells and in vivo endotoxemia model

• Choi Y
• Park SK
• Kim HM
• Kang JS
• Yoon YD
• Han SB
• Han JW
• Yang JS
• Han G

• KMID: 1104812
• DOI: http://doi.org/10.3858/emm.2008.40.5.574

Abstract

In light of the anti-inflammatory properties of histone deacetylase (HDAC) inhibitors, such as suberoylanilide hydroxamic acid (SAHA) and trichostatin A (TSA), we examined a new HDAC inhibitor KBH-A42 for its anti-inflammatory activities. KBH-A42 showed noteworthy anti-inflammatory properties in vitro via suppression of the production of TNF-alpha, a proinflammatory cytokine, and nitric oxide (NO), a proinflammatory effector molecule, in LPS-stimulated RAW264.7 cells and peritoneal macrophages. It also inhibited TNF-alpha production in vivo as demonstrated in a LPS-induced mouse endotoxemia model. The levels of TNF-alpha, IL-1beta, IL-6 and iNOS mRNAs determined by RT-PCR propose that the inhibition of these pro-inflammatory mediators by KBH-A42 resulted from inhibiting expression of these genes. However, the EMSA study to see the effect of KBH-A42 on the binding of NF-kappaB, a transcription factor, to a specific DNA sequence showed that the binding of NF-kappaB to DNA was not changed regardless of increasing the concentration of KBH-A42 in the presence and absence of LPS stimulation. Interestingly, DNA binding of another transcription factor AP-1 dose-dependently increased by KBH-A42. KBH-A42 differentially regulated the phosphorylation of MAP kinases. While the phosphprylation of ERK1/2 and SAPK/JNK was not affected by KBH-A42, the phosphorylation of p38 decreased by KBH-A42. These results showed that KBH-A42 inhibits production of proinflammatory cytokines in macrophages by decreasing their mRNA levels, and p38 kinase is involved in the KBH-A42-mediated inhibition.

Keyword

anti-inflammatory agents; histone deacetylases; NF-kappaB; nitric oxide; transcription factor AP-1; tumor necrosis factor-alpha; vorinostat

MeSH Terms

Animals
Blotting, Western
Cell Line
Cell Survival/drug effects
Cytokines/blood/genetics/*metabolism
Electrophoretic Mobility Shift Assay
Endotoxemia/blood/metabolism/pathology
Enzyme Inhibitors/chemistry/*pharmacology
Histone Deacetylases/*antagonists & inhibitors
Hydroxamic Acids/chemistry/*pharmacology
Interleukin-1beta/genetics/metabolism
Interleukin-6/genetics/metabolism
Macrophages/cytology/*drug effects/metabolism
Mice
Mitogen-Activated Protein Kinase 1/metabolism
Mitogen-Activated Protein Kinase 3/metabolism
Mitogen-Activated Protein Kinases/metabolism
Molecular Structure
NF-kappa B/metabolism
Nitric Oxide/metabolism
Nitric Oxide Synthase Type II/genetics/metabolism
Phosphorylation/drug effects
Piperidones/chemistry/*pharmacology
Protein Binding/drug effects
Reverse Transcriptase Polymerase Chain Reaction
Transcription Factor AP-1/metabolism
Tumor Necrosis Factor-alpha/blood/genetics/metabolism