Apoptotic signaling in methylglyoxal-treated human osteoblasts involves oxidative stress, c-Jun N-terminal kinase, caspase-3, and p21-activated kinase 2

J Cell Biochem. 2007 Mar 1;100(4):1056-69. doi: 10.1002/jcb.21114.

Abstract

Methylglyoxal (MG) is a reactive dicarbonyl compound endogenously produced mainly from glycolytic intermediates. MG is cytotoxic through induction of cell death, and elevated MG levels in diabetes patients are believed to contribute to diabetic complications. In this report, we show for the first time that MG treatment triggers apoptosis in human osteoblasts. We further show that MG-induced apoptosis of osteoblasts involves specific apoptotic biochemical changes, including oxidative stress, c-Jun N-terminal kinase (JNK) activation, mitochondrial membrane potential changes, cytochrome C release, increased Bax/Bcl-2 protein ratios, and activation of caspases (caspase-9, caspase-3) and p21-activated protein kinase 2 (PAK2). Treatment of osteoblasts with SP600125, a JNK-specific inhibitor, led to a reduction in MG-induced apoptosis and decreased activation of caspase-3 and PAK2, indicating that JNK activity is upstream of these events. Experiments using anti-sense oligonucleotides against PAK2 further showed that PAK2 activation is required for MG-induced apoptosis in osteoblasts. Interestingly, we also found that MG treatment triggered nuclear translocation of NF-kappaB, although the precise regulatory role of NF-kappaB activation in MG-induced apoptosis remains unclear. Lastly, we examined the effect of MG on osteoblasts in vivo, and found that exposure of rats to dietary water containing 100-200 microM MG caused bone mineral density (BMD) loss. Collectively, these results reveal for the first time that MG treatment triggers apoptosis in osteoblasts via specific apoptotic signaling, and causes BMD loss in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Bone Density / drug effects
  • Caspase 3 / metabolism*
  • Caspase 9 / metabolism
  • Cells, Cultured
  • Enzyme Activation / drug effects
  • Female
  • Humans
  • Immunoblotting
  • JNK Mitogen-Activated Protein Kinases / metabolism*
  • NF-kappa B / metabolism
  • Oligonucleotides, Antisense / genetics
  • Osteoblasts / cytology
  • Osteoblasts / drug effects*
  • Osteoblasts / metabolism
  • Oxidative Stress / drug effects
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / metabolism
  • Pyruvaldehyde / pharmacology*
  • Rats
  • Rats, Wistar
  • Reactive Oxygen Species / metabolism
  • Signal Transduction / drug effects
  • Transfection
  • p21-Activated Kinases

Substances

  • NF-kappa B
  • Oligonucleotides, Antisense
  • Reactive Oxygen Species
  • Pyruvaldehyde
  • PAK2 protein, human
  • Pak2 protein, rat
  • Protein-Serine-Threonine Kinases
  • p21-Activated Kinases
  • JNK Mitogen-Activated Protein Kinases
  • Caspase 3
  • Caspase 9