Impaired glycolytic metabolism causes chondrocyte hypertrophy-like changes via promotion of phospho-Smad1/5/8 translocation into nucleus

Osteoarthritis Cartilage. 2013 May;21(5):700-9. doi: 10.1016/j.joca.2013.01.013. Epub 2013 Feb 4.

Abstract

Objective: Hypertrophy-like changes are often observed in chondrocytes during the development of osteoarthritis (OA). These changes play a crucial part in the OA-associated cartilage degradation and osteophyte formation. However, the pathogenesis leading to such changes is still unknown. In this study, we investigated the mechanism by which these hypertrophy-like changes are induced from the viewpoint of impaired glycolytic metabolism.

Methods: The effect of sodium fluoride (NaF) on glycolytic metabolism of cultured chondrocytes was confirmed by measurement of intracellular adenosine triphosphate (ATP) production. Translocation of phosphorylated Smad1/5/8 to the nucleus was evaluated by subcellular fractionation and Western blotting. Chondrocyte hypertrophy-like changes were investigated by real-time RT-PCR and Western blot analysis of differentiation markers.

Results: ATP production was dose-dependently decreased by NaF in the human chondrocytic cell line HCS-2/8. In addition, both chondrocyte proliferation and differentiation were inhibited, whereas cell death was promoted by treatment with NaF. Interestingly, combinational treatment with NaF and lactate enhanced translocation of phospho-Smad1/5/8 to the nucleus, as well as gene expression of ALP, VEGF, COL10a1, and matrix metalloproteinase13 (MMP13), which were the markers of late mature and hypertrophic chondrocytes. Furthermore, the production of type X collagen and activation of MMP9 were also promoted under the same conditions.

Conclusions: These findings suggest that decreased ATP production by NaF promotes hypertrophy-like changes via activation of phospho-Smad1/5/8 in the presence of lactate. Novel metabolic aspects of OA pathogenesis are indicated herein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / biosynthesis
  • Bone Morphogenetic Proteins / metabolism
  • Cell Death / drug effects
  • Cell Differentiation / drug effects
  • Cell Nucleus / metabolism
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Chondrocytes / drug effects
  • Chondrocytes / metabolism*
  • Chondrocytes / pathology
  • Dose-Response Relationship, Drug
  • Drug Combinations
  • Glycolysis / drug effects
  • Glycolysis / physiology
  • Humans
  • Hypertrophy / chemically induced
  • Hypertrophy / metabolism
  • NF-kappa B / metabolism
  • Signal Transduction / drug effects
  • Smad Proteins, Receptor-Regulated / metabolism*
  • Smad1 Protein / metabolism
  • Smad5 Protein / metabolism
  • Smad8 Protein / metabolism
  • Sodium Fluoride / administration & dosage
  • Sodium Fluoride / pharmacology
  • Sodium Lactate / pharmacology

Substances

  • Bone Morphogenetic Proteins
  • Drug Combinations
  • NF-kappa B
  • SMAD1 protein, human
  • SMAD5 protein, human
  • SMAD9 protein, human
  • Smad Proteins, Receptor-Regulated
  • Smad1 Protein
  • Smad5 Protein
  • Smad8 Protein
  • Adenosine Triphosphate
  • Sodium Fluoride
  • Sodium Lactate