Figure 5
Effect of CQ and AQ on ROS production and apoptosis in hRPTCs under HG conditions. HKC-8 cells were incubated in 5 or 30 mM D-glucose for 24 hours and used for analysis. CQ (200 μM) or AQ (10 μM) were pretreated for 1 hour before exposure to 30 mM D-glucose for investigating effect of the drugs on hRPTCs under HG conditions. (A) Representative confocal images of MitoSox (red) and H2-DCFDA (green) showing that mitochondrial and intracellular ROS production was reduced in CQ- and AQ-treated hRPTCs under HG conditions. (B,C) Western blot analyses revealed that the expression of the apoptogenic proteins Cyt. C and Bax was decreased, while the expression of the anti-apoptotic protein Bcl2 was increased after CQ or AQ treatment under HG conditions. Band intensities representing Bcl-2, Bax, Cyt. C and GAPDH expression levels were converted into densitometry using ImageJ software in the ratio of Bcl2 to Bax and Cyt. C to GAPDH. Results are means ± SEM. for experiments in triplicate. (*p < 0.05 vs 5 mM, #p < 0.05, ##p < 0.01 vs 30 mM, Scale bar 10 μm).
