title: High glucose causes apoptosis in cultured human pancreatic islets of Langerhans: a potential role for regulation of specific Bcl family genes toward an apoptotic cell death program.
abstract: Type 2 diabetes is characterized by insulin resistance and inadequate insulin secretion. In the advanced stages of the disease, beta-cell dysfunction worsens and insulin therapy may be necessary to achieve satisfactory metabolic control. Studies in autopsies found decreased beta-cell mass in pancreas of people with type 2 diabetes. Apoptosis, a constitutive program of cell death modulated by the Bcl family genes, has been implicated in loss of beta-cells in animal models of type 2 diabetes. In this study, we compared the effect of 5 days' culture in high glucose concentration (16.7 mmol/l) versus normal glucose levels (5.5 mmol/l) or hyperosmolar control (mannitol 11 mmol/l plus glucose 5 mmol/l) on the survival of human pancreatic islets. Apoptosis, analyzed by flow cytometry and electron and immunofluorescence microscopy, was increased in islets cultured in high glucose (HG5) as compared with normal glucose (NG5) or hyperosmolar control (NG5+MAN5). We also analyzed by reverse transcriptase-polymerase chain reaction and Western blotting the expression of the Bcl family genes in human islets cultured in normal glucose or high glucose. The antiapoptotic gene Bcl-2 was unaffected by glucose change, whereas Bcl-xl was reduced upon treatment with HG5. On the other hand, proapoptotic genes Bad, Bid, and Bik were overexpressed in the islets maintained in HG5. To define the pancreatic localization of Bcl proteins, we performed confocal immunofluorescence analysis on human pancreas. Bad and Bid were specifically expressed in beta-cells, and Bid was also expressed, although at low levels, in the exocrine pancreas. Bik and Bcl-xl were expressed in other endocrine islet cells as well as in the exocrine pancreas. These data suggest that in human islets, high glucose may modulate the balance of proapoptotic and antiapoptotic Bcl proteins toward apoptosis, thus favoring beta-cell death.
authors.affiliation: Laboratory of Molecular Medicine, Department of Internal Medicine, University of Rome Tor Vergata, 00133 Rome, Italy. federicm@uniroma2.it
authors.forename: M
authors.initials: M
authors.lastname: Federici
authors.forename: M
authors.initials: M
authors.lastname: Hribal
authors.forename: L
authors.initials: L
authors.lastname: Perego
authors.forename: M
authors.initials: M
authors.lastname: Ranalli
authors.forename: Z
authors.initials: Z
authors.lastname: Caradonna
authors.forename: C
authors.initials: C
authors.lastname: Perego
authors.forename: L
authors.initials: L
authors.lastname: Usellini
authors.forename: R
authors.initials: R
authors.lastname: Nano
authors.forename: P
authors.initials: P
authors.lastname: Bonini
authors.forename: F
authors.initials: F
authors.lastname: Bertuzzi
authors.forename: L N
authors.initials: LN
authors.lastname: Marlier
authors.forename: A M
authors.initials: AM
authors.lastname: Davalli
authors.forename: O
authors.initials: O
authors.lastname: Carandente
authors.forename: A E
authors.initials: AE
authors.lastname: Pontiroli
authors.forename: G
authors.initials: G
authors.lastname: Melino
authors.forename: P
authors.initials: P
authors.lastname: Marchetti
authors.forename: R
authors.initials: R
authors.lastname: Lauro
authors.forename: G
authors.initials: G
authors.lastname: Sesti
authors.forename: F
authors.initials: F
authors.lastname: Folli
mesh_terms: D017209:Apoptosis / Q000235:genetics; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000502:physiology; D005947:Glucose / Q000008:administration & dosage* / Q000494:pharmacology; D006801:Humans; D007515:Islets of Langerhans / Q000187:drug effects* / Q000502:physiology*; D011519:Proto-Oncogenes / Q000502:physiology; D014018:Tissue Distribution; D014158:Transcription, Genetic / Q000502:physiology
chemical_list: D005947:Glucose