title: N-acetylcysteine protects pancreatic islet against glucocorticoid toxicity.
abstract: OBJECTIVE
  Reactive oxygen species (ROS) are involved in many physiological and pathological processes. In the present study, we analysed whether the synthetic glucocorticoid dexamethasone induces oxidative stress in cultured pancreatic islets and whether the effects of dexamethasone on insulin secretion, gene expression, and viability can be counteracted by concomitant incubation with N-acetylcysteine (NAC).
  
  
  METHODS
  ROS production was measured by dichlorofluorescein (DCFH-DA) assay, insulin secretion by radioimmunoassay, intracellular calcium dynamics by fura-2-based fluorescence, gene expression by real-time polymerase chain reaction analyses and cell viability by the MTS assay.
  
  
  RESULTS
  Dexamethasone (Dexa) increased ROS production and decreased glucose-stimulated insulin secretion after 72 hours incubation. Intracellular ROS levels were decreased and the insulin secretion capacity was recovered by concomitant treatment with Dexa+NAC. The total insulin content and intracellular Ca2+ levels were not modulated in either Dexa or Dexa+NAC groups. There was a decrease in the NAD(P)H production, used as an indicator of viability, after dexamethasone treatment. Concomitant incubation with NAC returned viability to control levels. Dexa also decreased synaptotagmin VII (SYT VII) gene expression. In contrast, the Dexa+NAC group demonstrated an increased expression of SYT VII compared to controls. Surprisingly, treatment with NAC decreased the gene expression of the antioxidant enzyme copper zinc superoxide dismutase soluble.
  
  
  CONCLUSIONS
  Our results indicate that dexamethasone increases ROS production, decreases viability, and impairs insulin secretion in pancreatic rat islets. These effects can be counteracted by NAC, which not only decreases ROS levels but also modulates the expression of genes involved in the secretory pathway and those coding for antioxidant enzymes.
authors: Department of Anatomy, Cellular Biology, Physiology and Biophysics, Institute of Biology, State University of Campinas, UNICAMP, Campinas, Brazil.
authors: Letícia P
authors: LP
authors: Roma
authors: Camila A M
authors: CA
authors: Oliveira
authors: Everardo M
authors: EM
authors: Carneiro
authors: Gabriela G
authors: GG
authors: Albuquerque
authors: Antonio C
authors: AC
authors: Boschero
authors: Kleber L A
authors: KL
authors: Souza
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003907:Dexamethasone / Q000037:antagonists & inhibitors* / Q000633:toxicity; D016166:Free Radical Scavengers / Q000494:pharmacology; D005609:Free Radicals / Q000378:metabolism; D005938:Glucocorticoids / Q000037:antagonists & inhibitors* / Q000633:toxicity; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects; D020011:Protective Agents / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism*; D050857:Synaptotagmins / Q000187:drug effects / Q000378:metabolism
chemical_list: D016166:Free Radical Scavengers; D005609:Free Radicals; D005938:Glucocorticoids; D007328:Insulin; D020011:Protective Agents; D017382:Reactive Oxygen Species; D050857:Synaptotagmins; D003907:Dexamethasone; D000111:Acetylcysteine