Figure 5
Short-term prometryn treatment induced oxidative-stress in mice liver. (a) Prometryn treatment significantly increased the protein expression of glutathione S-transferase alpha 1/2 (GSTA1/2) in mouse liver. Band signal intensities were analyzed by Image Lab® Software Version 12 and normalized with intensities obtained from loading control after staining with commercial Ponceau S for total protein. (b) Relative hydrogen peroxide (H2O2) levels of control and prometryn treated liver. (c) ROS formation was detected in HepG2 liver cells stained with a cell permeable fluorescent probe H2DCFDA and treated with 10–30 μM prometryn. H2O2 (200 μM) was used as a positive oxidative stress control. ROS levels were detected by fluorescence spectroscopy at an Ex-502 nm and Em-523 nm. (d) Elevated oxidative stress in HepG2 liver cells treated with prometryn was confirmed by staining with CellROX Deep Red. Meanwhile, 1 h pretreatment with mitochondrial-targeted antioxidant MitoTEMPO significantly decreased the oxidative stress. Shown are spectrofluorometric data expressing the fluorescence intensity of MitoSOX after treatment with prometryn and/or MitoTEMPO. PRO Prometryn, MT MitoTEMPO, MT + P MitoTEMPO + Prometryn. Bars represent the mean ± SEM; n = 4–8 mice per group. *p < 0.05, **p < 0.001, # = p**** < 0.0001.
