Glutathione maintenance mitigates age-related susceptibility to redox cycling agents.
Isolated hepatocytes from young (4-6mo) and old (24-26mo) F344 rats were exposed to increasing concentrations of menadione, a vitamin K derivative and redox cycling agent, to determine whether the age-related decline in Nrf2-mediated detoxification defenses resulted in heightened susceptibility to xenobiotic insult. An LC50for each age group was established, which showed that aging resulted in a nearly 2-fold increase in susceptibility to menadione (LC50for young: 405μM; LC50for old: 275μM). Examination of the known Nrf2-regulated pathways associated with menadione detoxification revealed, surprisingly, that NAD(P)H: quinone oxido-reductase 1 (NQO1) protein levels and activity were induced 9-fold and 4-fold with age, respectively (p=0.0019 and p=0.018; N=3), but glutathione peroxidase 4 (GPX4) declined by 70% (p=0.0043; N=3). These results indicate toxicity may stem from vulnerability to lipid peroxidation instead of inadequate reduction of menadione semi-quinone. Lipid peroxidation was 2-fold higher, and GSH declined by a 3-fold greater margin in old versus young rat cells given 300µM menadione (p<0.05 and p≤0.01 respectively; N=3). We therefore provided 400µMN-acetyl-cysteine (NAC) to hepatocytes from old rats before menadione exposure to alleviate limits in cysteine substrate availability for GSH synthesis during challenge. NAC pretreatment resulted in a >2-fold reduction in cell death, suggesting that the age-related increase in menadione susceptibility likely stems from attenuated GSH-dependent defenses. This data identifies cellular targets for intervention in order to limit age-related toxicological insults to menadione and potentially other redox cycling compounds.
Linus Pauling Institute, Oregon State University, Corvallis, OR 97331-6512, USA; Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331-7305, USA.
Nicholas O
NO
Thomas
Linus Pauling Institute, Oregon State University, Corvallis, OR 97331-6512, USA.
Kate P
KP
Shay
Linus Pauling Institute, Oregon State University, Corvallis, OR 97331-6512, USA; Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331-7305, USA.
Amanda R
AR
Kelley
Linus Pauling Institute, Oregon State University, Corvallis, OR 97331-6512, USA.
Judy A
JA
Butler
Linus Pauling Institute, Oregon State University, Corvallis, OR 97331-6512, USA; Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331-7305, USA. Electronic address: tory.hagen@oregonstate.edu.
Tory M
TM
Hagen
D000111:Acetylcysteine / Q000494:pharmacology; D000375:Aging / Q000378:metabolism*; D000818:Animals; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism*; D005979:Glutathione Peroxidase / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects*; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D051381:Rats; D011916:Rats, Inbred F344; D024483:Vitamin K 3 / Q000633:toxicity*
D024483:Vitamin K 3; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D005979:Glutathione Peroxidase; C523860:glutathione peroxidase 4, rat; D016660:NAD(P)H Dehydrogenase (Quinone); C507703:NQO1 protein, rat; D005978:Glutathione; D000111:Acetylcysteine
Aging; BSO (PubChem CID: 119565); DMF (PubChem CID: 6228); Detoxification Capacity; Glutathione; Menadione; Menadione (PubChem CID: 4055); NAC (PubChem CID: 12035); Redox-cycling
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