Toxic effects of buthionine sulfoximine on human hepatocytes L-02 through ROS/p38 pathway
WANG Yuan
GUAN Su
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.
Keywords：buthionine sulfoximineliver cellscell viabilityglutathione synthetasep38 proteinreactive oxidative species
Publication Date：2018-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 25-28 )