Mellado-Gil, Jose MAguilar-Diosdado, Manuel2012-10-222012-10-222005Mellado-Gil JM, Aguilar-Diosdado M. Assay for high glucose-mediated islet cell sensitization to apoptosis induced by streptozotocin and cytokines. Biol Proced Online; 2005, 7:162-711480-9222http://hdl.handle.net/10668/581Journal Article;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).enApoptosisDiabetes MellitusEstreptozotocinaCitocinasMedical Subject Headings::Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Death::ApoptosisMedical Subject Headings::Diseases::Nutritional and Metabolic Diseases::Metabolic Diseases::Glucose Metabolism Disorders::Diabetes MellitusMedical Subject Headings::Chemicals and Drugs::Organic Chemicals::Nitroso Compounds::Nitrosourea Compounds::StreptozocinMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Intercellular Signaling Peptides and Proteins::Cytokinesresearch article16281079open access10.1251/bpo113PMC1280327