Elevated glutathione is not sufficient to protect against doxorubicin-induced nuclear damage in heart in multidrug resistance-associated protein 1 (Mrp1/Abcc1) null mice.
Cardiotoxicity is a major dose-limiting adverse effect of doxorubicin (DOX), mediated in part by overproduction of reactive oxygen species and oxidative stress. Abcc1 (Mrp1) mediates the efflux of reduced and oxidized glutathione (GSH, GSSG) and is also a major transporter that effluxes the GSH conjugate of 4-hydroxy-2-nonenal (HNE; GS-HNE), a toxic product of lipid peroxidation formed during oxidative stress. To assess the role of Mrp1 in protecting the heart from DOX-induced cardiac injury, wild-type (WT) and Mrp1 null (Mrp1(-/-)) C57BL/6 littermate mice were administered DOX (15 mg/kg) or saline (7.5 ml/kg) i.v., and heart ventricles were examined at 72 hours. Morphometric analysis by electron microscopy revealed extensive injuries in cytosol, mitochondria, and nuclei of DOX-treated mice in both genotypes. Significantly more severely injured nuclei were observed in Mrp1(-/-) versus WT mice (P = 0.031). GSH and the GSH/GSSG ratio were significantly increased in treatment-naïve Mrp1(-/-) versus WT mice; GSH remained significantly higher in Mrp1(-/-) versus WT mice after saline and DOX treatment, with no changes in GSSG or GSH/GSSG. GS-HNE, measured by mass spectrometry, was lower in the hearts of treatment-naïve Mrp1(-/-) versus WT mice (P < 0.05). DOX treatment decreased GS-HNE in WT but not Mrp1(-/-) mice, so that GS-HNE was modestly but significantly higher in Mrp1(-/-) versus WT hearts after DOX. Expression of enzymes mediating GSH synthesis and antioxidant proteins did not differ between genotypes. Thus, despite elevated GSH levels in Mrp1(-/-) hearts, DOX induced significantly more injury in the nuclei of Mrp1(-/-) versus WT hearts.
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.).
Jun
J
Deng
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.).
Donna
D
Coy
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.).
Wei
W
Zhang
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.).
Manjula
M
Sunkara
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.).
Andrew J
AJ
Morris
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.).
Chi
C
Wang
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.).
Luksana
L
Chaiswing
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.).
Daret
D
St Clair
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.) maryv@email.uky.edu paiboon@fdu.edu.
Mary
M
Vore
Department of Toxicology and Cancer Biology (J.D., D.C., L.C., W.Z., D.St.C., M.V.), Division of Cardiovascular Medicine (M.S., A.J.M.), Markey Cancer Center (C.W.) University of Kentucky, Lexington, Kentucky; Department of Pathology and Laboratory Medicine, William S. Middleton Memorial Veterans Administration Hospital and University of Wisconsin Medical School, Madison Wisconsin (L.C.); and School of Pharmacy, Fairleigh Dickinson University, Florham Park, New Jersey (P.J.) maryv@email.uky.edu paiboon@fdu.edu.
Paiboon
P
Jungsuwadee
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D066126:Cardiotoxicity / Q000378:metabolism; D002467:Cell Nucleus / Q000187:drug effects*; D004317:Doxorubicin / Q000633:toxicity*; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism*; D019803:Glutathione Disulfide / Q000378:metabolism; D015227:Lipid Peroxidation; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics*; D009206:Myocardium / Q000378:metabolism* / Q000648:ultrastructure*; D018384:Oxidative Stress
D000903:Antibiotics, Antineoplastic; D004317:Doxorubicin; D005978:Glutathione; D019803:Glutathione Disulfide; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C524061:glutathionyl 4-hydroxy-2-nonenal conjugate; C421109:multidrug resistance-associated protein 1