Inhibition of HDAC6 attenuates LPS-induced inflammation in macrophages by regulating oxidative stress and suppressing the TLR4-MAPK/NF-κB pathways

Biomed Pharmacother. 2019 Sep:117:109166. doi: 10.1016/j.biopha.2019.109166. Epub 2019 Jun 27.

Abstract

Background: Histone deacetylase 6 (HDAC6) has been considered as an important regulator in the development of inflammatory diseases. However, the mechanism of HDAC6 in regulating inflammatory responses has not been fully determined. In the present study, we aim to investigate the role and mechanisms of HDAC6 in regulating inflammation in lipopolysaccharide (LPS)-activated macrophages.

Methods: Flow cytometry was used to determine a suitable treatment dosage of ACY-1215 on lipopolysaccharide (LPS)-activated macrophages for the present study. The RAW264.7 macrophages were divided into normal, LPS-treated, and ACY-1215 treated groups, respectively. For the ACY-1215 group, ACY-1215 (10 μM) was added to the medium 2 h prior to treatment with LPS (1 μg/ml) for 24 h. In this study, ROS, inflammatory cytokines, the ultrastructure of mitochondria, mitochondrial membrane potential, RNA and protein expression assay were detected respectively. Subsequently, the effect of HDAC6 knockdown on inflammatory response in LPS-activated RAW264.7 macrophages was also detected.

Results: Inhibition of HDAC6 inhibited the overproduction of ROS and suppressed the expression of pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 in LPS-activated RAW264.7 cells. Pretreatment with ACY-1215 could normalize the ultrastructure of mitochondria and mitochondrial membrane potential in LPS-activated macrophages. Moreover, the protein expression of TLR4, Nrf2, HO-1 and the activation of MAPK and NF-κB signaling pathways were normalized by the inhibition of HDAC6.

Conclusions: Inhibition of HDAC6 exhibited protective role against LPS-induced inflammation in RAW264.7 cells by regulating oxidative stress and suppressing the activation of TLR4- MAPK/NF-κB signaling pathway.

Keywords: ACY-1215; Histone deacetylase 6; Inflammation; Oxidative stress.

MeSH terms

  • Animals
  • Cytokines / metabolism
  • Heme Oxygenase-1 / metabolism
  • Histone Deacetylase 6 / metabolism*
  • Histone Deacetylase Inhibitors / pharmacology*
  • Hydroxamic Acids / pharmacology
  • Inflammation / pathology*
  • Inflammation Mediators / metabolism
  • Lipopolysaccharides
  • Macrophages / drug effects
  • Macrophages / metabolism
  • Macrophages / pathology*
  • Macrophages / ultrastructure
  • Membrane Potential, Mitochondrial / drug effects
  • Mice
  • Mitogen-Activated Protein Kinases / metabolism*
  • NF-E2-Related Factor 2 / metabolism
  • NF-kappa B / metabolism*
  • Oxidative Stress* / drug effects
  • Pyrimidines / pharmacology
  • RAW 264.7 Cells
  • Reactive Oxygen Species / metabolism
  • Signal Transduction / drug effects
  • Toll-Like Receptor 4 / metabolism*

Substances

  • Cytokines
  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • Inflammation Mediators
  • Lipopolysaccharides
  • NF-E2-Related Factor 2
  • NF-kappa B
  • Pyrimidines
  • Reactive Oxygen Species
  • Toll-Like Receptor 4
  • Heme Oxygenase-1
  • Mitogen-Activated Protein Kinases
  • Histone Deacetylase 6
  • ricolinostat