High-Dialysate-Glucose-Induced Oxidative Stress and Mitochondrial-Mediated Apoptosis in Human Peritoneal Mesothelial Cells
Figure 1
High-dialysate-glucose-induced mitochondrial dependent apoptosis in HPMCs. The HPMCs were isolated from human omentums and cultured at 4 concentrations of glucose, 5 mM (as a control), 84 mM, 138 mM, and 236 mM glucose for 24 h. (a) Exposure to high glucose resulted in a dose-dependent decrease in cellular viability by MTT assay. (b) Apoptotic cells were stained with annexin V-FITC and PI and detected using flow cytometry. (b) High-dialysate-glucose-induced cell apoptosis in both the early apoptotic cell (□, annexin V+/PI-) and the late apoptotic cells (■, annexin V+/PI+). (c) HPMCs were stained with annexin V-FITC (green signal) and DAPI (blue signal). Enhanced fluorescent green signals of the annexin V-FITC were observed in the HPMCs treated with high-dialysate glucose. (d-e) Analysis of cytochrome c release, caspase activation, and PARP cleavage of the high-glucose-treated HPMCs. Dose-dependent stimulation of cytochrome c released from the mitochondria into the cytosol was observed in the glucose-treated HPMCs (d). An increase in the cleavage of caspase-3 and caspase-9, and PARP was observed in the high-glucose-treated HPMCs (e). Plots represent the mean ± standard deviation (SD) of 4 independent experiments. Statistical significance: compared with the control; COXIV, cytochrome c oxidase subunit IV; PARP, poly(ADP-ribose) polymerase.
