摘要
Abstract
Objective To study the toxic effects and mechanism of buthionine sulfoximine(BSO)on human normal hepatocytes L-02 and its mechanism.Methods ①Human normal hepatocytes L-02 were incubated with 0,10,25,50, 100,and 200 μmol/L BSO for 48 h,the cell viability was measured by MTT and trypan blue staining.②L-02 cells were incubated with 0,10,25,50,100,and 200 μmol/L BSO for 48 h, the content of glutathione(GSH)was detected by DTNB method.③L-02 cells were incubated with 200 μmol/L BSO for 0,2,12,24,36,and 48 h,Western blotting was applied to detect the phosphorylation of p 38.④L-02 cells were incubated with 200 μmol/L BSO for 0,2,12,24,36, and 48 h,fluorescence probe H2DCFDA was used to observe the production of ROS in L-02 cells.⑤L-02 cells were di-vided into the control group,BSO group,SB203580 group,and BSO+SB203580 group.No drugs were added to the con-trol group,BSO group was given 200 μmol/L BSO,while the SB203580 group was added with 20 μmol/L SB203580,and the BSO+SB203580 group was first incubated with 20 μmol/L SB203580 for 15 min, then was added with 200 μmol/L BSO.After 24 h,the survival rate of each group was measured by MTT method,and the level of ROS was detected by fluo-rescence probe H2DCFDA.Results ①BSO reduced the survival rate of L-02 cells in a dose-dependent manner,and the minimum toxic dose was 50 μmol/L(P<0.05).②BSO could reduce the GSH content of L-02 cells in a dose-dependent manner,and it reached the minimum value since the concentration of BSO was 25 μmol/L.③BSO increased the phospho-rylation level of p38 in L-02 cells in a time-dependent manner(P<0.05).④Compared with control group, the level of ROS increased in L-02 cells when treated with BSO,and reached its peak at 24 h.⑤The ROS level in the BSO group was higher and the cell survival rate was lower than those of the control group(both P<0.05).There was no statistically sig-nificant difference in the ROS level and cell survival rate between the control group and SB 203580 group(P>0.05).The ROS level in the BSO+SB203580 group was lower and the cell survival rate was higher than those of the BSO group(both P<0.05).Conclusion BSO has certain cytotoxicity to human normal liver cells by increasing the phosphorylation level of p38 and ROS production,and decreasing the content of GSH.关键词
丁胱亚磺酰亚胺/肝细胞/细胞存活率/谷胱甘肽合成酶/p38蛋白/活性氧簇Key words
buthionine sulfoximine/liver cells/cell viability/glutathione synthetase/p38 protein/reactive oxidative species分类
医药卫生
