title: [Oxidative stress induces apoptosis via NF-kappaB-iNOS-nitric oxide pathway in pancreatic beta-cells].
abstract: OBJECTIVE
  To explore the possible mechanism of tert-butyl hydroperoxide (t-BHP)-induced apoptosis in murine MIN6 pancreatic beta-cells.
  
  
  METHODS
  MIN6 cells were cultured in vitro. Cell damage was evaluated by epifluorescence microscopy after staining with AO-EB. The percentage of cell apoptosis was determined by flow cytometric assay after Annexin- V-PI staining. Nitric oxide levels were measured by Griess assay. Inducible nitric oxide synthase(iNOS) protein and NF-kappaBp65 fragment were detected by Western blot.
  
  
  RESULTS
  Exposure of 25 micromol/L t-BHP to MIN6 cells for 60 min, cell viability was reduced and the percentage of apoptosis was increased significantly. The levels of cytoplasmic iNOS protein and nitrite were elevated. Meanwhile, treatment with t-BHP resulted in nucleus NF-kappaBp65 fragment peaking at 20 min. Both L-NAME and N-Acetyl-l-cysteine (NAC) attenuated the elevated levels of nitrite and percentage of apoptosis due to t-BHP alone.
  
  
  CONCLUSIONS
  NF-kappa-iNOS-nitric oxide signalling pathway can mediated t-BHP induced apoptosis in MIN6 cells .
authors.affiliation: Fujian Institute of Endocrinology, Union Hospital, Fujian Medical University, Fuzhou 350001, China.
authors.forename: Yan-ping
authors.initials: YP
authors.lastname: Wang
authors.forename: Xiao-dong
authors.initials: XD
authors.lastname: Pan
authors.forename: Su-yuan
authors.initials: SY
authors.lastname: Jiang
authors.forename: Li-bin
authors.initials: LB
authors.lastname: Liu
authors.forename: Zhou
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D050417:Insulin-Secreting Cells / Q000166:cytology*; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D018384:Oxidative Stress / Q000502:physiology*; D015398:Signal Transduction; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
chemical_list: D016328:NF-kappa B; D009569:Nitric Oxide; D020122:tert-Butylhydroperoxide; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse