Injuring Effect And Mechanism On Human Liver Cells L-02 Induced By Copper

Objective: To investigate the injury of human liver cell L-02 induced by copper, andresearch the mechanism from oxidized stress damaging to mitochondrion.Methods: L-02 cells were cultured in vitro, and exposed to various concentrations ofcopper sulfate(0,50,100,150,200μmol/L)for 18 hours;L-02 cells were treated respectively with 150μmol/L copper sulfate for 0,6,12,18,24 hours. The viability of cells and the function of mitochondria were determined by colorimetric 3-[4,5-dimethyl thiazol-2-yl]-2,5-diphenyltetrazolium bromide(MTT) assay. The activity of SOD was detected by xanthine oxidase method, and the content of GSH was measured by colorimetric and MDA was detected by Thiobituric acid reaction(TBA)method in supernatant fluid. Mitochondrion ultrastructure was observed by transmission electron microscope. Combine Rhodamine123(Rh123) and Propidium Iodide(PI) with Flow Cytometer to detect the change of mitochondrial membrane potential(MMP) and cell apoptosis. Cytochrome C(Cty C) releasing from mitochondrion to endochylema was observed by immunocytochemistry.Results:1. MTT results revealed Copper could inhibit the viability of cells and damage the function of mitochondrion in dose and time dependent (p<0. 05, p<0. 01).2. We observed the changes of the activity of SOD and the content of GSH and MDA of L-02 cell after incubated with copper . The results showed that copper could restrain the activity of SOD, and decrease the content of GSH and increase the level of MDA in supernatant fluid (p<0. 05, p<0. 01).