Modulation of hydrogen peroxide-induced DNA damage, MAPKs activation and cell death in PC12 by ergothioneine.
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.
Colognato
Renato
R
Dip. Scienze dell'Uomo e dell'Ambiente, Università di Pisa, Sezione di Genetica Via S. Giuseppe 22, 56126 Pisa, Italy.
Laurenza
Incoronata
I
Fontana
Ilaria
I
Coppedé
Fabio
F
Siciliano
Gabriele
G
Coecke
Sandra
S
Aruoma
Okezie I
OI
Benzi
Luca
L
Migliore
Lucia
L
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*
D000975:Antioxidants; D006861:Hydrogen Peroxide; D004880:Ergothioneine; D048051:p38 Mitogen-Activated Protein Kinases