Fig. 4

The protective effect of Zn against BHP-induced oxidative stress in hepatocytes. A The cell viability assessed by CCK8 in response to the BHP incubation at various concentrations. B–C The mitochondrial ROS (B) and MMP (C) in response to the incubation BHP at various concentrations. D Effect of Zn pretreatment on the level of intracellular ROS generation by 100 μmol/L BHP incubation. E Intracellular ROS generation was measured by DCFH-DA staining. F Effect of Zn pretreatment on intracellular MMP in hepatocytes subjected to 100 μmol/L BHP incubation. G Intracellular ROS was measured by JC-1 staining. In the presence of a high MMP, JC-1 forms J-aggregates that emit red fluorescence, while JC-1 monomeric form emits green fluorescence at low MMP. H–I Effect of Zn pretreatment on intracellular ATP content and mtDNA copy number of ATP6 in hepatocytes subjected to 100 μmol/L BHP incubation. K–L Effect of Zn pretreatment on MT4, Nrf-2, PGC-1α, PPAR-α protein expression in hepatocytes subjected to 100 μmol/L BHP incubation. Graph bars in A and B were analyzed using unpaired two-tailed Student's t-test (^∗P < 0.05), while graph bars in C–L marked with different letters on top represent statistically significant results (P < 0.05) based on Tukey's post hoc one-way ANOVA analysis, whereas bars labelled with the same letter correspond to results that show no statistically significant differences. Data are mean ± SEM, n = 4–6