title: Glucose and tolbutamide induce apoptosis in pancreatic beta-cells. A process dependent on intracellular Ca2+ concentration.
abstract: High concentrations of glucose are considered to be toxic for the pancreatic beta-cell. However, the mechanisms underlying beta-cell dysfunction and resulting cell death are not fully characterized. In the present study we have demonstrated that incubation of pancreatic islets and beta-cells from ob/ob mice and Wistar rats with glucose induced a process of apoptotic beta-cell death, as shown by DNA laddering, TdT-mediated dUTP-biotin nick end-labeling (TUNEL) technique, and by using DNA-staining dye HOECHST 33342. The obtained results show that the percentage of apoptotic cells was dependent on glucose concentration, being minimal at 11 mM glucose. At a concentration of 100 microM, aurintricarboxylic acid, an inhibitor of endonuclease activity, almost completely inhibited apoptosis triggered by 17 mM glucose. We have also shown that long term incubation with 100 microM sulfonylurea, tolbutamide, triggered apoptosis in pancreatic beta-cells. The process of beta-cell death induced by high glucose concentration and tolbutamide were Ca2+-dependent, because introduction to the culture medium of 50 microM D-600 or 200 microM diazoxide, which blocked glucose- and tolbutamide-induced [Ca2+]i increase, inhibited apoptosis. Thus, this study shows for the first time that high glucose concentrations and tolbutamide induce apoptosis in pancreatic beta-cells, and that this process is Ca2+-dependent.
authors.affiliation: The Rolf Luft Center for Diabetes Research, Department of Molecular Medicine, Karolinska Institute, Karolinska Hospital, S-171 76 Stockholm, Sweden.
authors.forename: I B
authors.initials: IB
authors.lastname: Efanova
authors.forename: S V
authors.initials: SV
authors.lastname: Zaitsev
authors.forename: B
authors.initials: B
authors.lastname: Zhivotovsky
authors.forename: M
authors.initials: M
authors.lastname: Köhler
authors.forename: S
authors.initials: S
authors.lastname: Efendić
authors.forename: S
authors.initials: S
authors.lastname: Orrenius
authors.forename: P O
authors.initials: PO
authors.lastname: Berggren
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology*; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D008820:Mice, Obese; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D051381:Rats; D017208:Rats, Wistar; D014044:Tolbutamide / Q000494:pharmacology*
chemical_list: D007004:Hypoglycemic Agents; D014044:Tolbutamide; D005947:Glucose; D002118:Calcium