title: Synergistic depletion of astrocytic glutathione by glucose deprivation and peroxynitrite: correlation with mitochondrial dysfunction and subsequent cell death.
abstract: Previously we reported that immunostimulated astrocytes were highly vulnerable to glucose deprivation. The augmented death was mimicked by the peroxynitrite (ONOO )-producing reagent 3-morpholinosydnonimine (SIN-1). Here we show that glucose deprivation and ONOO- synergistically deplete intracellular reduced glutathione (GSH) and augment the death of astrocytes via formation of cyclosporin A-sensitive mitochondrial permeability transition (MPT) pore. Astrocytic GSH levels were only slightly decreased by glucose deprivation or SIN-1 (200 microM) alone. In contrast, a rapid and large depletion of GSH was observed in glucose-deprived/ SIN-1-treated astrocytes. The depletion of GSH occurred before a significant release of lactate dehydrogenase (a marker of cell death). Superoxide dismutase and ONOO-scavengers completely blocked the augmented death, indicating that the reaction of nitric oxide with superoxide to form ONOO was implicated. Furthermore, nitrotyrosine immunoreactivity (a marker of ONOO-) was markedly enhanced in glucose-deprived/SIN-1 -treated astrocytes. Mitochondrial transmembrane potential (MTP) was synergistically decreased in glucose-deprived/SIN-1-treated astrocytes. The glutathione synthase inhibitor L-buthionine-(S,R)-sulfoximine markedly decreased the MTP and increased lactate dehydrogenase (LDH) releases in SIN-1-treated astrocytes. Cyclosporin A, an MPT pore blocker, completely prevented the MTP depolarization as well as the enhanced LDH releases in glucose-deprived/SIN-1-treated astrocytes.
authors.affiliation: Department of Pharmacology, College of Medicine, Ewha Women's University, Seoul, Republic of Korea.
authors.forename: C
authors.initials: C
authors.lastname: Ju
authors.forename: K N
authors.initials: KN
authors.lastname: Yoon
authors.forename: Y K
authors.initials: YK
authors.lastname: Oh
authors.forename: H C
authors.initials: HC
authors.lastname: Kim
authors.forename: C Y
authors.initials: CY
authors.lastname: Shin
authors.forename: J R
authors.initials: JR
authors.lastname: Ryu
authors.forename: K H
authors.initials: KH
authors.lastname: Ko
authors.forename: W K
authors.initials: WK
authors.lastname: Kim
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000378:metabolism* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D004357:Drug Synergism; D005947:Glucose / Q000172:deficiency*; D005978:Glutathione / Q000031:analogs & derivatives / Q000172:deficiency* / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000502:physiology; D008981:Molsidomine / Q000031:analogs & derivatives / Q000494:pharmacology; D009566:Nitrates / Q000494:pharmacology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009603:Nitroso Compounds / Q000494:pharmacology; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D026423:S-Nitroso-N-Acetylpenicillamine; D026422:S-Nitrosoglutathione
chemical_list: D009566:Nitrates; D020030:Nitric Oxide Donors; D009603:Nitroso Compounds; C039642:peroxynitric acid; D026422:S-Nitrosoglutathione; C002385:linsidomine; D026423:S-Nitroso-N-Acetylpenicillamine; D008981:Molsidomine; D005978:Glutathione; D010396:Penicillamine; D005947:Glucose