Journal Article

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

Ryuichi Nakahara, T. Machida, Keiichiro Nishida, Daisuke Kaneda, Masahiro Horita, M. Ozawa, Yoshihisa Nasu, Toshifumi Ozaki, Ayumu Takeshita, Aki Yoshida, Ryozo Harada

2017

Overview

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.

• Publisher: Springer Science and Business Media LLC,Springer International Publishing,Springer Nature B.V
• Subject: ADAM protein / ADAMTS4 Protein / ADAMTS4 Protein - genetics / ADAMTS5 Protein / ADAMTS5 Protein - genetics / Aggrecan / Aggrecans / Aggrecans - genetics / Allergology / Axial stress / Biomedical and Life Sciences / Biomedicine / Cartilage, Articular / Cartilage, Articular - cytology / Cascades / Cbfa-1 protein / Cells, Cultured / Chondrocytes / Chondrocytes - drug effects / Chondrocytes - metabolism / Collagen Type II / Collagen Type II - genetics / Collagenase 3 / Core Binding Factor Alpha 1 Subunit / Core Binding Factor Alpha 1 Subunit - genetics / Core Binding Factor Alpha 1 Subunit - metabolism / Cytokines / Cytokines - metabolism / Dermatology / Elongation / Enzyme-linked immunosorbent assay / Gene expression / Humans / Immunocytochemistry / Immunology / Inhibitors / Interleukin 6 / Janus kinase / Janus Kinase Inhibitors / Janus Kinase Inhibitors - pharmacology / Kinases / MAP kinase / Matrix metalloproteinase / Matrix Metalloproteinase 13 / Matrix Metalloproteinase 13 - genetics / Matrix metalloproteinases / Mechanical stimuli / Mitogen-Activated Protein Kinases / Mitogen-Activated Protein Kinases - metabolism / Neurology / Nuclear transport / Original Research Paper / Pharmacology/Toxicology / Phosphorylation / Piperidines / Piperidines - pharmacology / Polymerase chain reaction / Protein kinase / Pyrimidines / Pyrimidines - pharmacology / Pyrroles / Pyrroles - pharmacology / Rheumatology / Stat protein / Stat3 protein / STAT3 Transcription Factor / STAT3 Transcription Factor - metabolism / Stimulation / Strain / Stress, Mechanical / Thrombospondin / Transcription Factor RelA / Transcription Factor RelA - metabolism / Translocation / Tumor necrosis factor-α / Western blotting