High-Dialysate-Glucose-Induced Oxidative Stress and Mitochondrial-Mediated Apoptosis in Human Peritoneal Mesothelial Cells
Figure 3
High-dialysate-glucose-induced mitochondrial anomalies. (a,b) A fluoroprobe, MitoSOX Red (MitoSOX), was introduced for the selective detection of superoxides in the mitochondria of HPMCs and was measured using confocal microscopy and flow cytometry. The presence of fragmented mitochondria was prominent in the high-glucose-treated HPMCs. (b) A dose-dependent increase in mitochondrial superoxide production was observed in the high-glucose-treated HPMCs. (c) The shift of mitochondrial morphology from tubular to fragmented forms was observed in response to high-dialysate glucose stimulation. (d) Large-scale deletions of mtDNA were observed using long-range PCR in HPMCs stimulated by 5, 84, 138, and 236 mM glucose. Using the primer sets L8150/H16450, we detected large-scale deletions of mtDNA in HPMCs. The 8301 bp band was amplified from the wild-type mtDNA. Multiple mtDNA deletions (◄) were amplified from the HPMCs treated with 138 and 236 mM glucose. Lane M, 1 Kb DNA ladder. (e) The ultrastructure of mitochondria in HPMCs was observed using a transmission electron microscope. These mitochondria exhibited arch-shaped cristae and vacuolization in high-dialysate-glucose-treated HPMCs. Original magnification ×4000 and ×8000. The higher magnification field (×8000) is indicated by the square in the ×4000 image. Plots represent the mean ± SD from 4 independent experiments. Statistical significance: and compared with the control.
