Fig. 1: Decreased peroxidase activity of global hepatic PRDX in NASH.
a Peroxidase activity of global hepatic PRDX after normal chow (NC) or high-fat diet (HFD) feeding. 8-week-old male C57BL/6 mice were fed a NC or HFD for 18 weeks and liver samples were collected for biochemical analyses. Global hepatic PRDX peroxidase activity was measured using a classic Trx-TrxR-NADPH coupled assay. NADPH consumption was monitored via absorbance at 340 nm (A340) in 15 min assay duration. Meanwhile, the background activity was assessed without Trx and TrxR, but only with H2O2 and NADPH. To calculate the initial NADPH consumption rate (initial rate) (A340/min/protein (g)) in the first 5 min, a smooth curve was drawn through A340 readings, and the initial rate was calculated by performing a simple linear regression. Global PRDX peroxidase activity was calculated by subtracting the background activity (initial rate) from total activity (initial rate). n = 6 mice per group. b Protein levels of hepatic PRDX family enzymes after HFD (as in a) and quantitation. n = 6 mice per group; ns, no significance. c Peroxidase activity of global hepatic PRDX after NC or western diet (WD). 8-week-old male C57BL/6 mice were fed a NC or WD for 20 weeks and global hepatic PRDX peroxidase activity was measured using a classic Trx-TrxR-NADPH coupled assay. n = 6 mice per group. d Protein levels of hepatic PRDX family enzymes after NC or WD feeding (as in c) and quantitation. n = 6 mice per group; ns, no significance. e Peroxidase activity of global hepatic PRDX after NC or methionine and choline deficient diet (MCD). 8-week-old male C57BL/6 mice were fed a NC or MCD for 5 weeks and global hepatic PRDX peroxidase activity was measured using a classic Trx-TrxR-NADPH coupled assay. n = 6 mice per group. f Protein levels of hepatic PRDX family enzymes after NC or MCD feeding (as in e) and quantitation. n = 6 mice per group; ns, no significance. All data are presented as means ± SEM. Unpaired and two-tailed Student’s t test was performed for a–f.
