Extracellular HSP72 induces proinflammatory cytokines in human periodontal ligament fibroblast cells through the TLR4/NFκB pathway in vitro

Arch Oral Biol. 2017 Nov:83:181-186. doi: 10.1016/j.archoralbio.2017.07.021. Epub 2017 Jul 30.

Abstract

Objective: The aim of the present study was to examine the effect of extracellular heat shock protein (HSP) 72 on human periodontal ligament fibroblast cells (hPDLFs) in vitro.

Design: hPDLFs were stimulated by recombinant human HSP72 (rhHSP72). TAK-242 was used to inhibit toll-like receptor 4 (TLR4) activity. Interleukin (IL)-6, IL-8 and tumor necrosis factor (TNF)-α mRNA levels were analyzed by real-time PCR and protein levels were analyzed by enzyme-linked immunosorbent assay. p65/RelA phosphorylation was analyzed by western blot.

Results: IL-6, IL-8 and TNF-α mRNA and protein levels were significantly increased by rhHSP72 stimulation. These effects were inhibited by TAK-242 treatment. Additionally, p65/RelA phosphorylation was increased after 5-min rhHSP72 stimulation, which was inhibited by TAK-242 treatment.

Conclusion: Extracellular HSP72 induces proinflammatory cytokines through TLR4/NF-κB in hPDLFs.

Keywords: HSP72; IL-6; IL-8; PDL; TNF-α.

MeSH terms

  • Blotting, Western
  • Enzyme-Linked Immunosorbent Assay
  • Fibroblasts / metabolism*
  • HSP72 Heat-Shock Proteins / pharmacology*
  • Humans
  • In Vitro Techniques
  • Interleukin-6 / metabolism*
  • Interleukin-8 / metabolism*
  • NF-kappa B / antagonists & inhibitors*
  • Periodontal Ligament / cytology*
  • Phosphorylation
  • Real-Time Polymerase Chain Reaction
  • Toll-Like Receptor 4 / antagonists & inhibitors*
  • Tumor Necrosis Factor-alpha / metabolism*

Substances

  • HSP72 Heat-Shock Proteins
  • Interleukin-6
  • Interleukin-8
  • NF-kappa B
  • TLR4 protein, human
  • Toll-Like Receptor 4
  • Tumor Necrosis Factor-alpha