Hyaluronan suppresses mechanical stress-induced expression of catabolic enzymes by human chondrocytes via inhibition of IL-1β production and subsequent NF-κB activation

Masatsugu Ozawa, Keiichiro Nishida, Aki Yoshida, Taichi Saitou, Ryozo Harada, Takahiro Machida, Toshihumi Ozaki

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Objective: To investigate the inhibitory effect of hyaluronan (HA) on mechanical stress- induced expression of a disintegrin and metalloproteinase with thrombospondin type1 motifs (ADAMTS)-4, -5 and matrix metalloproteinase (MMP)-13 by human chondrocytes.Materials and methods: Normal human articular chondrocytes were pre-incubated with or without 1.0 mg/mL HA (2700 kDa) for 12 h at 37 °C in stretch chambers, then they were exposed to uni-axial cyclic tensile strain (CTS, 0.5 Hz, 10 % elongation). The expression of ADAMTS-4, -5, and MMP-13 were analyzed by real-time polymerase chain reaction and Immunocytochemistry. The concentration of IL-1β in the supernatant was measured using enzyme-linked immunosorbent assay (ELISA). The nuclear translocation of runt-related transcription factor 2 (RUNX-2) and nuclear factor-κB (NF-κB) was examined by ELISA and immunocytochemistry, and phosphorylation of NF-κB was examined by western blotting.Results: HA inhibited mRNA expression of ADAMTS-4, -5, and MMP13 after 24 h CTS via inhibition of IL-1β secretion and NF-κB activation. However, HA failed to inhibit CTS-induced RUNX-2 expression and subsequent expression of ADAMTS-5 and MMP-13 1 h after CTS.Conclusions: Our results demonstrated that HA significantly suppressed mechanical stress-induced expression of catabolic proteases by inhibition of the NF-κB–IL-1β pathway, but did not suppress mechanical stress-induced RUNX-2 signaling.

Original languageEnglish
Pages (from-to)243-252
Number of pages10
JournalInflammation Research
Volume64
Issue number3-4
DOIs
Publication statusPublished - 2015

Fingerprint

Mechanical Stress
Hyaluronic Acid
Chondrocytes
Interleukin-1
Matrix Metalloproteinase 13
Enzymes
Enzyme-Linked Immunosorbent Assay
Immunohistochemistry
Thrombospondins
Disintegrins
Metalloproteases
Real-Time Polymerase Chain Reaction
Peptide Hydrolases
Transcription Factors
Joints
Western Blotting
Phosphorylation
Messenger RNA

Keywords

  • Aggrecanase
  • Chondrocyte
  • Hyaluronan
  • Mechanical stress
  • NF-κB
  • RUNX-2

ASJC Scopus subject areas

  • Pharmacology
  • Immunology

Cite this

@article{a32eb8c7ae6b4191a8bd0a527cd5e51e,
title = "Hyaluronan suppresses mechanical stress-induced expression of catabolic enzymes by human chondrocytes via inhibition of IL-1β production and subsequent NF-κB activation",
abstract = "Objective: To investigate the inhibitory effect of hyaluronan (HA) on mechanical stress- induced expression of a disintegrin and metalloproteinase with thrombospondin type1 motifs (ADAMTS)-4, -5 and matrix metalloproteinase (MMP)-13 by human chondrocytes.Materials and methods: Normal human articular chondrocytes were pre-incubated with or without 1.0 mg/mL HA (2700 kDa) for 12 h at 37 °C in stretch chambers, then they were exposed to uni-axial cyclic tensile strain (CTS, 0.5 Hz, 10 {\%} elongation). The expression of ADAMTS-4, -5, and MMP-13 were analyzed by real-time polymerase chain reaction and Immunocytochemistry. The concentration of IL-1β in the supernatant was measured using enzyme-linked immunosorbent assay (ELISA). The nuclear translocation of runt-related transcription factor 2 (RUNX-2) and nuclear factor-κB (NF-κB) was examined by ELISA and immunocytochemistry, and phosphorylation of NF-κB was examined by western blotting.Results: HA inhibited mRNA expression of ADAMTS-4, -5, and MMP13 after 24 h CTS via inhibition of IL-1β secretion and NF-κB activation. However, HA failed to inhibit CTS-induced RUNX-2 expression and subsequent expression of ADAMTS-5 and MMP-13 1 h after CTS.Conclusions: Our results demonstrated that HA significantly suppressed mechanical stress-induced expression of catabolic proteases by inhibition of the NF-κB–IL-1β pathway, but did not suppress mechanical stress-induced RUNX-2 signaling.",
keywords = "Aggrecanase, Chondrocyte, Hyaluronan, Mechanical stress, NF-κB, RUNX-2",
author = "Masatsugu Ozawa and Keiichiro Nishida and Aki Yoshida and Taichi Saitou and Ryozo Harada and Takahiro Machida and Toshihumi Ozaki",
year = "2015",
doi = "10.1007/s00011-015-0804-2",
language = "English",
volume = "64",
pages = "243--252",
journal = "Inflammation Research",
issn = "1023-3830",
publisher = "Birkhauser Verlag Basel",
number = "3-4",

}

TY - JOUR

T1 - Hyaluronan suppresses mechanical stress-induced expression of catabolic enzymes by human chondrocytes via inhibition of IL-1β production and subsequent NF-κB activation

AU - Ozawa, Masatsugu

AU - Nishida, Keiichiro

AU - Yoshida, Aki

AU - Saitou, Taichi

AU - Harada, Ryozo

AU - Machida, Takahiro

AU - Ozaki, Toshihumi

PY - 2015

Y1 - 2015

N2 - Objective: To investigate the inhibitory effect of hyaluronan (HA) on mechanical stress- induced expression of a disintegrin and metalloproteinase with thrombospondin type1 motifs (ADAMTS)-4, -5 and matrix metalloproteinase (MMP)-13 by human chondrocytes.Materials and methods: Normal human articular chondrocytes were pre-incubated with or without 1.0 mg/mL HA (2700 kDa) for 12 h at 37 °C in stretch chambers, then they were exposed to uni-axial cyclic tensile strain (CTS, 0.5 Hz, 10 % elongation). The expression of ADAMTS-4, -5, and MMP-13 were analyzed by real-time polymerase chain reaction and Immunocytochemistry. The concentration of IL-1β in the supernatant was measured using enzyme-linked immunosorbent assay (ELISA). The nuclear translocation of runt-related transcription factor 2 (RUNX-2) and nuclear factor-κB (NF-κB) was examined by ELISA and immunocytochemistry, and phosphorylation of NF-κB was examined by western blotting.Results: HA inhibited mRNA expression of ADAMTS-4, -5, and MMP13 after 24 h CTS via inhibition of IL-1β secretion and NF-κB activation. However, HA failed to inhibit CTS-induced RUNX-2 expression and subsequent expression of ADAMTS-5 and MMP-13 1 h after CTS.Conclusions: Our results demonstrated that HA significantly suppressed mechanical stress-induced expression of catabolic proteases by inhibition of the NF-κB–IL-1β pathway, but did not suppress mechanical stress-induced RUNX-2 signaling.

AB - Objective: To investigate the inhibitory effect of hyaluronan (HA) on mechanical stress- induced expression of a disintegrin and metalloproteinase with thrombospondin type1 motifs (ADAMTS)-4, -5 and matrix metalloproteinase (MMP)-13 by human chondrocytes.Materials and methods: Normal human articular chondrocytes were pre-incubated with or without 1.0 mg/mL HA (2700 kDa) for 12 h at 37 °C in stretch chambers, then they were exposed to uni-axial cyclic tensile strain (CTS, 0.5 Hz, 10 % elongation). The expression of ADAMTS-4, -5, and MMP-13 were analyzed by real-time polymerase chain reaction and Immunocytochemistry. The concentration of IL-1β in the supernatant was measured using enzyme-linked immunosorbent assay (ELISA). The nuclear translocation of runt-related transcription factor 2 (RUNX-2) and nuclear factor-κB (NF-κB) was examined by ELISA and immunocytochemistry, and phosphorylation of NF-κB was examined by western blotting.Results: HA inhibited mRNA expression of ADAMTS-4, -5, and MMP13 after 24 h CTS via inhibition of IL-1β secretion and NF-κB activation. However, HA failed to inhibit CTS-induced RUNX-2 expression and subsequent expression of ADAMTS-5 and MMP-13 1 h after CTS.Conclusions: Our results demonstrated that HA significantly suppressed mechanical stress-induced expression of catabolic proteases by inhibition of the NF-κB–IL-1β pathway, but did not suppress mechanical stress-induced RUNX-2 signaling.

KW - Aggrecanase

KW - Chondrocyte

KW - Hyaluronan

KW - Mechanical stress

KW - NF-κB

KW - RUNX-2

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U2 - 10.1007/s00011-015-0804-2

DO - 10.1007/s00011-015-0804-2

M3 - Article

C2 - 25693597

AN - SCOPUS:84925497184

VL - 64

SP - 243

EP - 252

JO - Inflammation Research

JF - Inflammation Research

SN - 1023-3830

IS - 3-4

ER -