Figure 6. Ceramide increases reactive oxygen species accumulation and lipid peroxidation in bovine lens epithelial cells

A: BLECs were loaded with DCFH-DA at final concentration of 50 μM for 60 min. After incubation, DCFH-DA was removed, and the cells were treated with C2-ceramide (30 μM) for 30-270 min as described in Methods. The control group was treated with vehicle after loading with DCFH-DA. Data shown are mean±SEM (n=8) percentages of DCF fluorescence normalized to the control (no C2-ceramide) at each sampling time. B: C2-ceramide induces production of TBARS in BLECs. The cells were incubated in complete DMEM containing C2-ceramide for 24 h. Cells were washed with PBS, and TBARS assay was carried out as described in Methods. The TBARS concentration was expressed as malondialdehyde (MDA) equivalents and normalized to protein amount in the cells. Results are the average±SEM of three experiments. The asterisk denotes a p<0.05.