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Assay for high glucose-mediated islet cell sensitization to apoptosis induced by streptozotocin and cytokines

Abstract

Pancreatic β-cell apoptosis is known to participate in the β-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)-1β, tumor necrosis factor (TNF)-α and interferon (IFN)-γ]-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).

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Correspondence to Manuel Aguilar-Diosdado.

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Mellado-Gil, J.M., Aguilar-Diosdado, M. Assay for high glucose-mediated islet cell sensitization to apoptosis induced by streptozotocin and cytokines. Biol. Proced. Online 7, 162–171 (2005). https://doi.org/10.1251/bpo113

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  • DOI: https://doi.org/10.1251/bpo113

Indexing terms

  • Apoptosis
  • Diabetes Mellitus
  • Streptozotocin
  • Cytokines