Experimental Design

The hepatocytes were grown in 6-well cell culture plates. After the hepatocytes density reached 80%, the cells were harvested and treated by adding FE with or without ucOCN (Mybiosource, San Diego, CA) into the DMEM for studying the effect of ucOCN on chicken embryonic hepatocytes. There were 5 groups: control group (CONT), FE group (FE, 10% FE, v/v), FE with ucOCN 1 ng/mL (FE-LOCN), 3 ng/mL (FE-MOCN), and 9 ng/mL (FE-HOCN). SP600125 (a JNK inhibitor, Beyotime, Shanghai, China) and NAC (a ROS scavenger, Beyotime, Shanghai, China) were used for investigating the effects of FE on the function of ROS-JNK signal pathway of the hepatocytes. Based on our pilot study, in which 0.5, 1, and 2 mM NAC and 2.5, 5, and 10 μM SP600125 were used, in this study, 2 mM NAC and 5 μM SP600125 were added with 10% FE at the same time in the cell culture fluid for testing the effects on FE on the function of ROS-JNK signal pathway. The number of hepatocytes, cell ultra-microstructure, viability, and apoptosis were detected after 48 h treatment, and protein expressions and enzyme concentrations were detected after 72 h treatment.