H2O2 alters metabolism in TM4 Sertoli cells in the mouse
WANG Chao
ZHAO Ang
FAN Li
MA Bo
SHANG Xue-jun
ZHANG Qi
Abstract：Objective:To explore the mechanisms of oxidative stress-induced damage to TM4 Sertoli cells in the mouse using metabolomics techniques based on gas chromatography-mass spectrometry (GC-MS).Methods:We established the model of oxidative stress-induced damage to mouse TM4 Sertoli cells by treatment with H2 O2.Then,we detected the survival rate and apoptosis rate of the TM4 cells by MTT and flow cytometry respectively,measured the concentration of ROS in the TM4 cells with the DCFH-DA fluorescent probe,and determined the levels of endogenous metabolites in the TM4 cells by GC-MS after H2 O2 intervention.Results:After 2 hours of treatment with H2O2 at 600 μmol/L,the survival rate of the TM4 cells was reduced to about 50％,and the total apoptosis rates in the low-(100 μmol/L),medium-(300 μmol/L),and high-dose (600 μmol/L) groups were (19.45 ± 0.53),(20.12 ± 0.58),and (37.13 ± 0.35)％,respectively,increased in a dose-dependent manner as compared with (10.28 ± 0.35)％ in the blank control (P ＜ 0.05).The ROS level was significantly higher in the medium-and high-dose groups than in the control ([1.27 ± 0.10] vs [1.00 ± 0.08]％,P ＜0.05;[2.07 ± 0.09] vs [1.00 ± 0.08]％,P ＜0.01).Compared with the blank control group,the high-dose H2 O2 group showed evident changes in the levels of amino acid and carbohydrates in the TM4 cells,more significantly in the levels of valine,norvaline,leucine,glutamic acid,arabinose,fructose,and 5-serotonin cholesterol (VIP ＞ 1,P ＜0.05).Conclusion:Oxidative stress-induced damage and apoptosis of TM4 Sertoli cells are closely associated with the metabolism of amino acid,glucose,and energy in the cells.
Keywords：TM4 Sertoli cellH2O2oxidative stressmetabolomicsgas chromatography-mass spectrometrymouse
Publication Date：2017-06-02
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 497-502 )