| Objective: To explore the effects of LY-EC on oxidative damage, energy metabolism of mitochondria and activities of enzyme in L-02 hepatocytes.Methods: The effects of LY-EC on cell survival rate were assessed by the reductions of tetrazolium dye (MTT) in cultured L-02 hepatocytes. The concentrations of LY-EC at above 60% survival rate were selected for the following tests. L-02 hepatocytes in all tests were incubated with 100mg/L, 25mgμmol/, 6.25mg/L, 1.56mg/L, 0.39mg/L concentrations of LY-EC for 12h according to cell survival rate. The levels of malonic aldehyde(MDA) and the glutathione (GSH), the activities of superoxide dismutase (SOD), succinate dehydrogenase(SDH), ATPase, glutathione transferase (GST)and the mitochondrial permeability transition pore(PTP) were measured by colorimetry method. The levels of cellular ATP, ADP, AMP content as well as the changes of ATP/ADP ratio and the energy charge(Ec) were detected by high performance liquid chromatography (HPLC),while the ultrastructure of mitochondria was observed under the electron microscope.Results: LY-EC induced severe oxidative damage in cultured L-02 hepatocytes. A significant negative correlation was found between the concentrations and the survival rates of L-02 hepatocytes treated with LY-EC(r=-0.939). Compared with the normal control group, the activities of superoxide dismutase(SOD), succinate dehydrogenase(SDH), ATPase and glutathione transferase(GST) in L-02 hepatocytes treated with LY-EC were significantly decreased(p<0.05), the open of mitochondrial permeability transition pore(PTP) and the levels of malonic aldehyde(MDA) were significantly increased(p<0.05), and the relationships between the concerntrations and the effects were observed among LY-EC-treated groups. The contents of glutathione(GSH) and total adenine nucleotide (TAN) and ATP/ADP ratio were significantly decreased(p<0.05), and the energy charge (E_c) were significantly increased(p<0.05) in hepatocytes teated with LY-EC compared with control group, but the obvious relationships between the concemtrations and the effects were not observed among LY-EC-treated groups. The changes in the ultrastructure of mitochondria and cytoplasm were observed under electro-microscopic at the highest concentration of LY-EC.Conclusions: LY-EC can obviously result in the decreases ofsurvival rate, oxidative damage and the inhibition of mitochondrialenergy metabolism in L-02 hepatocytes . |
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