TitleAscorbic acid promotes detoxification and elimination of 4-hydroxy-2(E)-nonenal in human monocytic THP-1 cells.
Publication TypeJournal Article
Year of Publication2009
AuthorsMiranda CL, Reed RL, Kuiper HC, Alber S, Stevens JF
JournalChem Res Toxicol
Date Published2009 May
KeywordsAldehydes, Apoptosis, Ascorbic Acid, Biological Transport, Caspase 3, Fluoresceins, Glutathione, Humans, Inactivation, Metabolic, Leukemia, Protein Carbonylation, Tumor Cells, Cultured

4-Hydroxy-2(E)-nonenal (HNE), a reactive aldehyde derived from oxidized lipids, has been implicated in the pathogenesis of cardiovascular and neurological diseases, in part by its ability to induce oxidative stress and by protein carbonylation in target cells. The effects of intracellular ascorbic acid (vitamin C) on HNE-induced cytotoxicity and protein carbonylation were investigated in human THP-1 monocytic leukemia cells. HNE treatment of these cells resulted in apoptosis, necrosis, and protein carbonylation. Ascorbic acid accumulated in the cells at concentrations of 6.4 or 8.9 mM after treatment with 0.1 or 1 mM ascorbate in the medium for 18 h. Pretreatment of cells with 1.0 mM ascorbate decreased HNE-induced formation of reactive oxygen species and formation of protein carbonyls. The protective effects of ascorbate were associated with an increase in the formation of GSH-HNE conjugate and its phase 1 metabolites, measured by LC-MS/MS, and with increased transport of GSH conjugates from the cells into the medium. Ascorbate pretreatment enhanced the efflux of the multidrug resistant protein (MRP) substrate, carboxy-2',7'-dichlorofluorescein (CDF), and it prevented the HNE-induced inhibition of CDF export from THP-1 cells, suggesting that the protective effect of ascorbate against HNE cytotoxicity is through modulation of MRP-mediated transport of GSH-HNE conjugate metabolites. The formation of ascorbate adducts of HNE was observed in the cell exposure experiments, but it represented a minor pathway contributing to the elimination of HNE and to the protective effects of ascorbate.

Alternate JournalChem. Res. Toxicol.
PubMed ID19326901
PubMed Central IDPMC2730585
Grant ListS10 RR022589 / RR / NCRR NIH HHS / United States
P30ES000210 / ES / NIEHS NIH HHS / United States
S10RR022589 / RR / NCRR NIH HHS / United States
R01 HL081721 / HL / NHLBI NIH HHS / United States
S10 RR022589-01 / RR / NCRR NIH HHS / United States
P30 ES000210-40 / ES / NIEHS NIH HHS / United States
R01 HL081721-03 / HL / NHLBI NIH HHS / United States
R01HL081721 / HL / NHLBI NIH HHS / United States
P30 ES000210 / ES / NIEHS NIH HHS / United States