title: Menadione-induced reactive oxygen species generation via redox cycling promotes apoptosis of murine pancreatic acinar cells.
abstract: Oxidative stress may be an important determinant of the severity of acute pancreatitis. One-electron reduction of oxidants generates reactive oxygen species (ROS) via redox cycling, whereas two-electron detoxification, e.g. by NAD(P)H:quinone oxidoreductase, does not. The actions of menadione on ROS production and cell fate were compared with those of a non-cycling analogue (2,4-dimethoxy-2-methylnaphthalene (DMN)) using real-time confocal microscopy of isolated perfused murine pancreatic acinar cells. Menadione generated ROS with a concomitant decrease of NAD(P)H, consistent with redox cycling. The elevation of ROS was prevented by the antioxidant N-acetyl-l-cysteine but not by the NADPH oxidase inhibitor diphenyliodonium. DMN produced no change in reactive oxygen species per se but significantly potentiated menadione-induced effects, probably via enhancement of one-electron reduction, since DMN was found to inhibit NAD(P)H:quinone oxidoreductase detoxification. Menadione caused apoptosis of pancreatic acinar cells that was significantly potentiated by DMN, whereas DMN alone had no effect. Furthermore, bile acid (taurolithocholic acid 3-sulfate)-induced caspase activation was also greatly increased by DMN, whereas DMN had no effect per se. These results suggest that acute generation of ROS by menadione occurs via redox cycling, the net effect of which is induction of apoptotic pancreatic acinar cell death. Two-electron detoxifying enzymes such as NAD(P)H:quinone oxidoreductase, which are elevated in pancreatitis, may provide protection against excessive ROS and exert an important role in determining acinar cell fate.
authors.affiliation: MRC Secretory Research Group, Department of Physiology and Division of Surgery and Oncology, University of Liverpool, Liverpool L69 3BX, United Kingdom. criddle@liverpool.ac.uk
authors.forename: David N
authors.initials: DN
authors.lastname: Criddle
authors.forename: Stuart
authors.initials: S
authors.lastname: Gillies
authors.forename: Heidi K
authors.initials: HK
authors.lastname: Baumgartner-Wilson
authors.forename: Mohammed
authors.initials: M
authors.lastname: Jaffar
authors.forename: Edwin C
authors.initials: EC
authors.lastname: Chinje
authors.forename: Sarah
authors.initials: S
authors.lastname: Passmore
authors.forename: Michael
authors.initials: M
authors.lastname: Chvanov
authors.forename: Stephanie
authors.initials: S
authors.lastname: Barrow
authors.forename: Oleg V
authors.initials: OV
authors.lastname: Gerasimenko
authors.forename: Alexei V
authors.initials: AV
authors.lastname: Tepikin
authors.forename: Robert
authors.initials: R
authors.lastname: Sutton
authors.forename: Ole H
authors.initials: OH
authors.lastname: Petersen
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000037:antagonists & inhibitors / Q000502:physiology; D009249:NADP / Q000378:metabolism; D009252:NADPH Dehydrogenase / Q000037:antagonists & inhibitors / Q000502:physiology; D010084:Oxidation-Reduction; D010179:Pancreas / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D024483:Vitamin K 3 / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D017382:Reactive Oxygen Species; D009249:NADP; D024483:Vitamin K 3; D016660:NAD(P)H Dehydrogenase (Quinone); C491732:Nqo1 protein, mouse; D009252:NADPH Dehydrogenase