Objective: To investigate the effects of 10% Liuyangmycin emulsifiable (LY-EC) on mitochondrial function in L-02 hepatocytes.
Methods: To detect the effect of different concentration of LY-EC (0 -200 mg/L) on L-02 hepatocytes survival rate with MTT method, and choose proper LY-EC treated concentrations (survival rate > 70%) for the following experiments. After hepatocytes were treated with LY-EC at 0.39, 1.56, 6.25 and 25.00 mg/L respectively for 12 h, MDA and GSH contents, SOD, SDH, ATPase activities were respectively analyzed by their assay kits using colorimetry. Then mitochondrial PTP open percentage and the contents of ATP, ADP and AMP were measured with HPLC assays, and ATP/ADP ratio and energy charge (EC) values were calculated. The effects of LY-EC on mitochondrial ultra-structures were observed by transmission electron microscopy.
Results: LY-EC induced the decrease of cell survival rates in a concentrate- dependent manner (r = 0.939, P < 0.05). Compared with control group, GSH contents and SOD, SDH and ATPase activities in groups treated with LY-EC decreased significantly, MDA contents and mitochondrial PTP open percentages increased (P < 0.05), the levels of cellular ATP, TAN and ATP/ADP ratio in L-02 hepatocytes decreased, while energy charge (EC) increased significantly (P < 0.05). Hepatocytes treated with LY-EC were observed cytoplasmic edema, mitochondria swelling and the deformation of mitochondrial cristae with transmission electron microscope.
Conclusion: LY-EC induced oxidative stress, mitochondrial function damage and energy metabolism disorder in L-02 hepatocytes.