Hydrogen peroxide inhibits formation of cartilage in chicken micromass cultures and decreases the activity of calcineurin: implication of ERK1/2 and Sox9 pathways

Róza Zákány; Zsolt Szíjgyártó; Csaba Matta; Tamás Juhász; Csilla Csortos; Kornélia Szucs; Gabriella Czifra; Tamás Bíró; László Módis; Pál Gergely

doi:10.1016/j.yexcr.2004.12.016

Hydrogen peroxide inhibits formation of cartilage in chicken micromass cultures and decreases the activity of calcineurin: implication of ERK1/2 and Sox9 pathways

Exp Cell Res. 2005 Apr 15;305(1):190-9. doi: 10.1016/j.yexcr.2004.12.016.

Authors

Róza Zákány¹, Zsolt Szíjgyártó, Csaba Matta, Tamás Juhász, Csilla Csortos, Kornélia Szucs, Gabriella Czifra, Tamás Bíró, László Módis, Pál Gergely

Affiliation

¹ Department of Anatomy, Histology and Embryology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary.

PMID: 15777799
DOI: 10.1016/j.yexcr.2004.12.016

Abstract

Calcineurin was found as a positive regulator of chondrogenesis in chondrifying chicken micromass cultures (HDCs), as cyclosporine A (CsA) reduced both the amount of cartilage and the expression of mRNAs of aggrecan and the chondrogenic transcription factor Sox9. Cartilage formation was inhibited by H(2)O(2) in a concentration-dependent manner without loss of cellular viability or severe decrease of cell number. Expression of both the mRNA and the unphosphorylated protein Sox9 was decreased, while its phosphorylation was stimulated by either H(2)O(2) or CsA. Oxidative stress decreased the activity of calcineurin but the phosphorylation of the member of MAPK family ERK1/2 was extremely elevated either by 1 mM H(2)O(2) or 2 muM CSA. The ERK inhibitor PD098059 attenuated the depletion of cartilage matrix as well as decreased the expression and phosphorylation of Sox9 in cultures treated with H(2)O(2) or CsA. Our results suggest that the chondrogenesis-inhibiting effect of H(2)O(2) is mediated, at least partly, by inhibition of calcineurin and by activation of ERK1/2. We also propose a regulatory role of calcineurin in the phosphorylation level of either ERK1/2 or Sox9 and a positive role of ERK1/2 in regulating both the expression level and the phosphorylation state of Sox9 in chicken HDCs.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Calcineurin / genetics
Calcineurin / physiology*
Calcineurin Inhibitors
Cartilage / embryology*
Chick Embryo
Chondrocytes / drug effects
Chondrocytes / physiology
Cyclosporine / pharmacology
Enzyme Inhibitors / pharmacology
Flavonoids / pharmacology
Gene Expression Regulation, Developmental / drug effects
High Mobility Group Proteins / genetics
High Mobility Group Proteins / metabolism*
Hydrogen Peroxide / pharmacology*
Limb Buds / physiology
Mitogen-Activated Protein Kinase 1 / antagonists & inhibitors
Mitogen-Activated Protein Kinase 1 / genetics
Mitogen-Activated Protein Kinase 1 / metabolism*
Mitogen-Activated Protein Kinase 3 / antagonists & inhibitors
Mitogen-Activated Protein Kinase 3 / genetics
Mitogen-Activated Protein Kinase 3 / metabolism*
Organ Culture Techniques
Reverse Transcriptase Polymerase Chain Reaction
SOX9 Transcription Factor
Transcription Factors / genetics
Transcription Factors / metabolism*

Substances

Calcineurin Inhibitors
Enzyme Inhibitors
Flavonoids
High Mobility Group Proteins
SOX9 Transcription Factor
Transcription Factors
Cyclosporine
Hydrogen Peroxide
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
Calcineurin
2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one