TY - JOUR
T1 - Effect of sulforaphane on glutathione-adduct formation and on glutathione_S_transferase-dependent detoxification of acrylamide in Caco-2 cells
AU - Pernice, Rita
AU - Hauder, Johanna
AU - Koehler, Peter
AU - Vitaglione, Paola
AU - Fogliano, Vincenzo
AU - Somoza, Veronika
PY - 2009/12
Y1 - 2009/12
N2 - The toxicity of dietary acrylamide (AA) depends on its biotransformation pathways, in which phase I cytochrome P-450 enzymes transform AA into glycidamide. The phase II enzyme glutathione_S_transferase (GST) catalyses the conjugation of AA with glutathione (GSH). GST induction by phytochemicals like sulforaphane (SFN) plays a role in chemoprevention. Here, the effect of SFN on the detoxification of AA through GSH conjugation was studied in Caco-2 cells. GSH adducts with AA and SFN were synthesized, identified by NMR and quantified by LC-MS/MS. Caco-2 cells were treated with either 2.5mM AA, 10 mM SFN or the combination of both for 24h. Concentrations of GSH conjugates (GSH-AA, GSH-SFN, SFN-GSH-AA), AA and SFN were analysed by LC-MS/MS. GSH contents and GST activity were determined photometrically. GST activity was increased after treatment of the cells with SFN (3876%, pr0.05) or AA (2574%, pr0.05). GSH concentrations decreased after all treatments. Quantitative data of GSH adduct formation showed that the reaction between GSH and SFN is favoured over that between GSH and AA. The data suggest that SFN might impair the GSH-dependent detoxification of AA by SFN-GSH adduct formation and, thus, lower the GSH concentrations available for its reaction with AA.
AB - The toxicity of dietary acrylamide (AA) depends on its biotransformation pathways, in which phase I cytochrome P-450 enzymes transform AA into glycidamide. The phase II enzyme glutathione_S_transferase (GST) catalyses the conjugation of AA with glutathione (GSH). GST induction by phytochemicals like sulforaphane (SFN) plays a role in chemoprevention. Here, the effect of SFN on the detoxification of AA through GSH conjugation was studied in Caco-2 cells. GSH adducts with AA and SFN were synthesized, identified by NMR and quantified by LC-MS/MS. Caco-2 cells were treated with either 2.5mM AA, 10 mM SFN or the combination of both for 24h. Concentrations of GSH conjugates (GSH-AA, GSH-SFN, SFN-GSH-AA), AA and SFN were analysed by LC-MS/MS. GSH contents and GST activity were determined photometrically. GST activity was increased after treatment of the cells with SFN (3876%, pr0.05) or AA (2574%, pr0.05). GSH concentrations decreased after all treatments. Quantitative data of GSH adduct formation showed that the reaction between GSH and SFN is favoured over that between GSH and AA. The data suggest that SFN might impair the GSH-dependent detoxification of AA by SFN-GSH adduct formation and, thus, lower the GSH concentrations available for its reaction with AA.
KW - Acrylamide
KW - Caco-2 cells
KW - GSH adducts
KW - Glutathione-S-transferase
KW - Sulforaphane
UR - https://www.scopus.com/pages/publications/71149113401
U2 - 10.1002/mnfr.200900447
DO - 10.1002/mnfr.200900447
M3 - Article
C2 - 19904761
AN - SCOPUS:71149113401
SN - 1613-4125
VL - 53
SP - 1540
EP - 1550
JO - Molecular Nutrition and Food Research
JF - Molecular Nutrition and Food Research
IS - 12
ER -