title: Assay for high glucose-mediated islet cell sensitization to apoptosis induced by streptozotocin and cytokines.
abstract: Pancreatic beta-cell apoptosis is known to participate in the beta-cell destruction process that occurs in diabetes. It has been described that high glucose level induces a hyperfunctional status which could provoke apoptosis. This phenomenon is known as glucotoxicity and has been proposed that it can play a role in type 1 diabetes mellitus pathogenesis. In this study we develop an experimental design to sensitize pancreatic islet cells by high glucose to streptozotocin (STZ) and proinflammatory cytokines [interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma]-induced apoptosis. This method is appropriate for subsequent quantification of apoptotic islet cells stained with Tdt-mediated dUTP Nick-End Labeling (TUNEL) and protein expression assays by Western Blotting (WB).
authors.affiliation: Endocrinology Service and Research Unit, Puerta del Mar Hospital, Cadiz, Spain.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Mellado-Gil
authors.forename: Manuel
authors.initials: M
authors.lastname: Aguilar-Diosdado