title: [Oxidative damage to the endoplasmic reticulum stress pathway of apoptosis-related molecules expression in MIN6 cell].
abstract: OBJECTIVE
  Through a third-butyl hydrogen peroxide (t-BHP) induced apoptosis in pancreatic islet β-cells to study the oxidative damage induced endoplasmic reticulum stress-JNK pathway of apoptosis related molecules in vitro.
  
  
  METHODS
  Mouse insulinoma(MIN6) cells was administered with t-BHP which were cultured in vitro. Choosing medicine with different concentrations(0-400 μmol/L)and time periods(0-8 h)to establish the cells apoptosis model. The percentage of cell viability was determined through CCK-8 assay. The percentage of apoptosis was determined through flow cytometric assay after Annexin-V-FITC-PI staining. The activity of caspase-3 was measured by the caspase-3 activity assay kit. The expression of Endoplasmic reticulum stress-related molecules and the apoptosis signal pathway IRE1, JNK, P-JNK, Caspase-3 were detected by Western blot.
  
  
  RESULTS
  The percentage of MIN6 cell viability was reducing with the concentration of t-BHP increasing. The Caspase-3 significantly change the activity after exposured of t-BHP in a concentration ≥ 25 μmol/L when the role of ≥1 h, With t-BHP concentration was increased, the role of prolonged, endoplasmic reticulum stress transmembrane protein IRE1α, P-JNK, active caspase-3 expression was significantly increased.
  
  
  CONCLUSIONS
  The study demonstrates that the percentage of MIN6 cell viability was reduced in a dose-dependent manner. Continuous exposuring of t-BHP induced oxidative damage in MIN6 cells to endoplasmic reticulum stress and apoptosis. The expression of Endoplasmic reticulum stress and apoptosis pathway-related molecules in cell apoptosis in a dose and time-dependent.
authors.affiliation: Fujian Institute of Endocrinology, the Affiliated Union Hospital of Fujian Medical University, Fuzhou 350001, China. wjjia81@hotmail.com
authors.forename: Wen-jia
authors.initials: WJ
authors.lastname: Chen
authors.forename: Xiao-ying
authors.initials: XY
authors.lastname: Liu
authors.forename: Lin-Xi
authors.initials: LX
authors.lastname: Wang
authors.forename: Yan-ping
authors.initials: YP
authors.lastname: Wang
authors.forename: Xiao-hong
authors.initials: XH
authors.lastname: Liu
authors.forename: Li-bin
authors.initials: LB
authors.lastname: Liu
mesh_terms: D000818:Animals; D051379:Mice; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D004722:Endoribonucleases / Q000187:drug effects / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism; D007340:Insulinoma / Q000378:metabolism; D048670:MAP Kinase Kinase 4 / Q000187:drug effects / Q000378:metabolism; D008565:Membrane Proteins / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D010749:Phosphoprotein Phosphatases / Q000187:drug effects / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction; D020122:tert-Butylhydroperoxide / Q000494:pharmacology; D054637:Dual-Specificity Phosphatases
chemical_list: D053148:Caspase 3; D004722:Endoribonucleases; C403982:ERN2 protein, human; D048670:MAP Kinase Kinase 4; D008565:Membrane Proteins; D010749:Phosphoprotein Phosphatases; D017346:Protein Serine-Threonine Kinases; D020122:tert-Butylhydroperoxide; C522152:Dusp22 protein, mouse; D054637:Dual-Specificity Phosphatases