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  • Title: [Oxidative damage to the endoplasmic reticulum stress pathway of apoptosis-related molecules expression in MIN6 cell].
    Author: Chen WJ, Liu XY, Wang LX, Wang YP, Liu XH, Liu LB.
    Journal: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi; 2011 Mar; 27(3):249-52, 256. PubMed ID: 21419040.
    Abstract:
    AIM: 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. CONCLUSION: 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.
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