title: Metformin prevents an adaptive increase in GSH and induces apoptosis under the conditions of GSH deficiency in H4IIE cells.
abstract: The antidiabetic effect of metformin is mediated by activation of AMP-activated kinase (AMPK). This study investigated whether metformin at a high pharmacologic concentration alters the levels of cellular GSH in H4IIE hepatocytes and if so, whether the agent affects cell viability under GSH-deficient conditions. Treatment of cells with either metformin or 5-aminoimidazole-4-carboxamide riboside (AICAR) increased dichlorofluorescein oxidation, as did tert-butylhydroxyquinone (t-BHQ). Metformin or AICAR treatment blocked a rebound increase in GSH produced by t-BHQ and decreased GSH content below that of control. Exposure of cells to metformin or metformin + t-BHQ for 24 hr did not produce cell death. However, metformin treatment in combination with t-BHQ for a prolonged period of time (48 hr) at the concentrations, at which each agent was non-toxic, produced apoptosis. Treatment of AICAR with t-BHQ resulted in similar effects. Induction of apoptosis by the combination treatment was evidenced by changes in mitochondrial cytochrome c content, BCl(xl) expression, poly(ADP-ribose)polymerase (PARP) cleavage and caspase-3 activation. Compound C, an AMPK inhibitor, reversed apoptosis and changes in the apoptotic markers, suggesting a role of AMPK activation by metformin in the apoptotic process. Similarly, metformin treatment, when combined with buthionine sulfoximine or doxorubicin, induced apoptosis. Our data indicated that metformin prevents an adaptive increase in cellular GSH in H4IIE cells, and therefore induces apoptosis under the conditions of GSH deficiency.
authors.affiliation: College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea.
authors.forename: Eun Ju
authors.initials: EJ
authors.lastname: Bae
authors.forename: Min Joo
authors.initials: MJ
authors.lastname: Cho
authors.forename: Sang Geon
authors.initials: SG
authors.lastname: Kim
mesh_terms: D055372:AMP-Activated Protein Kinases; D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line; D005978:Glutathione / Q000187:drug effects* / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects*; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D008687:Metformin / Q000494:pharmacology*; D009097:Multienzyme Complexes / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D017346:Protein Serine-Threonine Kinases / Q000187:drug effects / Q000378:metabolism
chemical_list: D007004:Hypoglycemic Agents; D009097:Multienzyme Complexes; D008687:Metformin; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases; D005978:Glutathione