title: Modulation of hydrogen peroxide-induced DNA damage, MAPKs activation and cell death in PC12 by ergothioneine.
abstract: OBJECTIVE
  Ergothioneine (EGT) is a natural occurring compound, synthesized by soil bacteria in fungal substrates, exhibiting antioxidant functions in many cell models. The aim of this study was to assess the effect of EGT in the prevention of H2O2-dependent cell death and oxidative damage on a model of neural cell derived from rat pheocromocytoma, the PC12.
  
  
  METHODS
  The ability of EGT was tested by the 3 (4,5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide (MTT) assay and Comet assay. H2O2 insult was challenged with increasing concentration of antioxidant using two different incubation periods: 1 and 23 h of EGT pre-treatment followed by 23 and 1 h of H2O2, respectively, for both the MTT and the Comet assay data.
  
  
  CONCLUSIONS
  The pre-treatment for 23 h with EGT, 250 microM and 1mM, followed by 1h of H2O2 incubation at the concentration of 250 and 500 microM, resulted in increased cell viability (P < 0.001) compared to the H2O2 cell batch. This correlated with a decrease in DNA damage as visualized by the Comet assay. Moreover, protein analysis reveals that in the presence of 250 microM of H2O2, EGT acted as a p38-MAPK and Akt specific inhibitor. EGT may play a protective role in rescuing cells from stress-induced apoptosis, likely by activating an intracellular antioxidant pathway involving p38 MAPK genes cascade.
authors.affiliation: Dip. Scienze dell'Uomo e dell'Ambiente, Università di Pisa, Sezione di Genetica Via S. Giuseppe 22, 56126 Pisa, Italy.
authors.forename: Renato
authors.initials: R
authors.lastname: Colognato
authors.forename: Incoronata
authors.initials: I
authors.lastname: Laurenza
authors.forename: Ilaria
authors.initials: I
authors.lastname: Fontana
authors.forename: Fabio
authors.initials: F
authors.lastname: Coppedé
authors.forename: Gabriele
authors.initials: G
authors.lastname: Siciliano
authors.forename: Sandra
authors.initials: S
authors.lastname: Coecke
authors.forename: Okezie I
authors.initials: OI
authors.lastname: Aruoma
authors.forename: Luca
authors.initials: L
authors.lastname: Benzi
authors.forename: Lucia
authors.initials: L
authors.lastname: Migliore
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects*; D020552:Comet Assay; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004880:Ergothioneine / Q000494:pharmacology*; D006861:Hydrogen Peroxide / Q000633:toxicity; D015151:Immunoblotting; D010084:Oxidation-Reduction; D016716:PC12 Cells; D051381:Rats; D013997:Time Factors; D048051:p38 Mitogen-Activated Protein Kinases / Q000187:drug effects / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D006861:Hydrogen Peroxide; D004880:Ergothioneine; D048051:p38 Mitogen-Activated Protein Kinases