High molecular weight hyaluronic acid regulates MMP13 expression in chondrocytes via DUSP10/MKP5.
To determine the effect of high molecular weight hyaluronic acid (HA) on matrix metalloproteinase 13 (MMP13) expression induced by tumor necrosis factor α (TNF-α) in chondrocytes. Human chondrocytic C28/I2 cells were incubated with TNF-α and HA. In some experiments, the cells were pre-incubated with a CD44 function-blocking monoclonal antibody (CD44 mAb) prior to addition of TNF-α and HA. The expression of MMP13 was determined by real-time reverse-transcription polymerase chain reaction (RT-PCR) and an enzyme linked immunosorbent assay, while the phosphorylation of signaling molecules was measured by western blot analysis. The transcriptional activity of activator protein 1 (AP-1) was analyzed by a reporter assay. To further clarify the molecular mechanisms of HA in MMP13 regulation, the expression level of dual-specificity protein phosphatase 10 (DUSP10)/mitogen-activated protein kinases phosphatase 5 (MKP5) in HA-treated chondrocytes was assessed by real-time RT-PCR, western blotting, and immunofluorescence microscopy. HA decreased MMP13 mRNA and protein expression induced by TNF-α. Blockage of HA-CD44 binding by CD44 mAb suppressed HA-mediated inhibition of MMP13. HA inhibited transient phosphorylation of p38 mitogen-activated protein kinase (MAPK) and c-jun NH2-terminal kinase (JNK) induced by TNF-α. Reporter assay findings also revealed that pre-treatment with HA inhibited the transcriptional activity of AP-1 mediated by TNF-α. Moreover, HA induced the expression of DUSP10/MKP5, a negative regulator of p38 MAPK and JNK pathways. These results indicate that HA-CD44 interactions downregulate TNF-α-induced MMP13 expression via regulation of DUSP10/MKP5, suggesting that HA plays an important role as a regulatory factor in cartilage degradation. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:331-339, 2017.
Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University, Kitakyushu, Fukuoka, Japan.|Division of Oral and Maxillofacial Surgery, Department of Science of Physical Function, Kyushu Dental University, Kitakyushu, Fukuoka, Japan.
Junya
J
Furuta
Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University, Kitakyushu, Fukuoka, Japan.
Wataru
W
Ariyoshi
Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University, Kitakyushu, Fukuoka, Japan.
Toshinori
T
Okinaga
Pharmaceuticals Information Group, Seikagaku Corporation, Tokyo, Japan.
Jun
J
Takeuchi
Division of Oral and Maxillofacial Surgery, Department of Science of Physical Function, Kyushu Dental University, Kitakyushu, Fukuoka, Japan.
Sho
S
Mitsugi
Division of Oral and Maxillofacial Surgery, Department of Science of Physical Function, Kyushu Dental University, Kitakyushu, Fukuoka, Japan.
Kazuhiro
K
Tominaga
Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University, Kitakyushu, Fukuoka, Japan.
Tatsuji
T
Nishihara
D002460:Cell Line; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism; D054637:Dual-Specificity Phosphatases / Q000378:metabolism*; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D006820:Hyaluronic Acid / Q000494:pharmacology* / Q000627:therapeutic use; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D053509:Matrix Metalloproteinase 13 / Q000378:metabolism*; D054639:Mitogen-Activated Protein Kinase Phosphatases / Q000378:metabolism*; D010003:Osteoarthritis / Q000188:drug therapy; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha; D055675:Viscosupplements / Q000494:pharmacology* / Q000627:therapeutic use; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D018808:Transcription Factor AP-1; D014409:Tumor Necrosis Factor-alpha; D055675:Viscosupplements; D006820:Hyaluronic Acid; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C120407:DUSP10 protein, human; D054639:Mitogen-Activated Protein Kinase Phosphatases; D054637:Dual-Specificity Phosphatases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13
DUSP10/MKP5; MAPKs; MMP13; chondrocyte; hyaluronic acid