Inhibitory effect of JAK inhibitor on mechanical stress-induced protease expression by human articular chondrocytes

Inflamm Res. 2017 Nov;66(11):999-1009. doi: 10.1007/s00011-017-1083-x. Epub 2017 Jul 27.

Abstract

Objective: To investigate whether janus kinase (JAK) inhibitor exhibits a chondro-protective effect against mechanical stress-induced expression of a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) and matrix metalloproteinase (MMPs) in human chondrocytes.

Materials and methods: Normal human articular chondrocytes were seeded onto stretch chambers and incubated with or without tofacitinib (1000 nM) for 12 h before mechanical stimulation or cytokine stimulation. Uni-axial cyclic tensile strain (CTS) (0.5 Hz, 10% elongation, 30 min) was applied and the gene expression levels of type II collagen α1 chain (COL2A1), aggrecan (ACAN), ADAMTS4, ADAMTS5, MMP13, and runt-related transcription factor 2 (RUNX-2) were examined by real-time polymerase chain reaction. Nuclear translocation of RUNX-2 and nuclear factor-κB (NF-κB) was examined by immunocytochemistry, and phosphorylation of mitogen-activated protein kinase (MAPK) and signaling transducer and activator of transcription (STAT) 3 was examined by western blotting. The concentration of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α in the supernatant was examined by enzyme-linked immunosorbent assay.

Results: COL2A1 and ACAN gene expression levels were decreased by CTS, but these catabolic effects were canceled by tofacitinib. Tofacitinib significantly down-regulated CTS-induced expression of ADAMTS4, ADAMTS5, MMP13, and RUNX2, and the release of IL-6 in supernatant by chondrocytes. Tofacitinib also reduced CTS-induced nuclear translocation of RUNX-2 and NF-κB, and phosphorylation of MAPK and STAT3.

Conclusion: Tofacitinib suppressed mechanical stress-induced expression of ADAMTS4, ADAMTS5, and MMP13 by human chondrocytes through inhibition of the JAK/STAT and MAPK cascades.

Keywords: Chondrocyte; Janus kinase; Mechanical stress; Rheumatoid arthritis; Tofacitinib.

MeSH terms

  • ADAMTS4 Protein / genetics
  • ADAMTS5 Protein / genetics
  • Aggrecans / genetics
  • Cartilage, Articular / cytology
  • Cells, Cultured
  • Chondrocytes / drug effects*
  • Chondrocytes / metabolism
  • Collagen Type II / genetics
  • Core Binding Factor Alpha 1 Subunit / genetics
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Cytokines / metabolism
  • Humans
  • Janus Kinase Inhibitors / pharmacology*
  • Matrix Metalloproteinase 13 / genetics
  • Mitogen-Activated Protein Kinases / metabolism
  • Piperidines / pharmacology*
  • Pyrimidines / pharmacology*
  • Pyrroles / pharmacology*
  • STAT3 Transcription Factor / metabolism
  • Stress, Mechanical*
  • Transcription Factor RelA / metabolism

Substances

  • ACAN protein, human
  • Aggrecans
  • COL2A1 protein, human
  • Collagen Type II
  • Core Binding Factor Alpha 1 Subunit
  • Cytokines
  • Janus Kinase Inhibitors
  • Piperidines
  • Pyrimidines
  • Pyrroles
  • RUNX2 protein, human
  • STAT3 Transcription Factor
  • Transcription Factor RelA
  • tofacitinib
  • Mitogen-Activated Protein Kinases
  • ADAMTS5 Protein
  • ADAMTS5 protein, human
  • MMP13 protein, human
  • Matrix Metalloproteinase 13
  • ADAMTS4 Protein
  • ADAMTS4 protein, human