Fig. 3.

BBD-serum reduces ethanol-mediated AML-12 cell damage. Cell death measurement was performed with Annexin V staining. a AML-12 cells were treated with different concentrations (100 nM–1000 mM) of ethanol for 24 h. b AML-12 cells were pretreated with different concentrations of normal serum/BBD-serum for 1 h followed by administering with ethanol (400 mM) for 24 h. c DCFH-DA staining was utilized to detect intracellular ROS generation. The cells pretreated with 10% BBD-serum for 1 h was subsequently added 400 mM ethanol and then were incubated with 10 μM DCFH-DA for an additional 45 min at 37 °C. Fluorescence microscope analysis were performed to identify cellular ROS activity. d, e Effects of 10% BBD-serum on CYP2E1 expression in transcriptional and protein level when cells suffered 400 mM ethanol. f Influence of BBD-serum on MDA, GSH and SOD changes were presented. Cells were incubated with 400 mM ethanol in presence of 10% BBD-serum. Data present are from three replicates as mean ± SD. ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 compared to control; *P < 0.05, **P < 0.01 compared to 10%N + EtOH group. N normal-serum, B BBD-serum, EtOH ethanol