Abeta[25-35] peptide and iron promote apoptosis in lymphocytes by an oxidative stress mechanism: involvement of H2O2, caspase-3, NF-kappaB, p53 and c-Jun

Neurotoxicology. 2002 Sep;23(3):351-65. doi: 10.1016/s0161-813x(02)00081-5.

Abstract

The Abeta deposition in the neuritic plaques is one of the major neuropathological hallmarks of the Alzheimer disease (AD). Studies in vitro have demonstrated that the Abeta[25-35] fragment, which contains the cytotoxic functional sequence of the amyloid peptide, induces neurotoxicity and cell death by apoptosis. Despite intense investigations, a complete picture of the precise molecular cascade leading to cell death in a single cellular model is still lacking. In this study, we provide evidence that Abeta[25-35] induce apoptosis either alone or in presence of iron in peripheral blood lymphocytes cells (PBL) in a concentration-dependent fashion by an oxidative stress mechanism involving: (1) the production of hydrogen peroxide (H2O2), reflected by rhodamine-positive fluorescent cells, (2) activation and/or translocation of NF-kappaB, p53 and c-Jun transcription factors showed by immunocytochemical diaminobenzidine positive nuclei, (3) activation of NF-kappaB complex by electrophoretic mobility shift assay/immuno-blotting/and ammonium pyrrolidinedithiocarbamate (PDTC) inhibition, (4) caspase-3 activation, reflected by caspase Ac-DEVD-cho inhibition, (5) mRNA synthesis de novo according to actinomycin D cell death inhibition. These results are consistent with the notion that the Abeta[25-35]/H2O2 generation precede the apoptotic process and that once H2O2 is generated, it is able to trigger a specific cell death signalisation. Thus, taken together these results, we present a well-ordered cascade of the major molecular events leading PBL to apoptosis. These results may contribute to explain the importance of Abeta alone or in the presence of redox-available iron in association with Abeta plaques (and neurofibrillary tangles) in AD brains and the significant role played by H2O2 as a second messenger of death signal in some degenerative diseases linked to oxidative stress stimuli.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Amyloid beta-Peptides / pharmacology*
  • Apoptosis / drug effects*
  • Blotting, Western
  • Caspase 3
  • Caspases / genetics
  • Caspases / physiology*
  • Cell Death / drug effects
  • Electrophoretic Mobility Shift Assay
  • Genes, jun / genetics
  • Genes, jun / physiology*
  • Genes, p53 / genetics
  • Genes, p53 / physiology*
  • Humans
  • Hydrogen Peroxide / metabolism*
  • Immunohistochemistry
  • In Vitro Techniques
  • Iron / pharmacology*
  • Lymphocytes / drug effects*
  • Lymphocytes / metabolism
  • Male
  • NF-kappa B / genetics
  • NF-kappa B / physiology*
  • Oxidative Stress / drug effects*
  • Peptide Fragments / pharmacology*
  • RNA, Messenger / biosynthesis
  • Reactive Oxygen Species / metabolism

Substances

  • Amyloid beta-Peptides
  • NF-kappa B
  • Peptide Fragments
  • RNA, Messenger
  • Reactive Oxygen Species
  • amyloid beta-protein (25-35)
  • Hydrogen Peroxide
  • Iron
  • CASP3 protein, human
  • Caspase 3
  • Caspases