H₂O₂ alters metabolism in TM4 Sertoli cells in the mouse.
- Author:
Chao WANG
1
;
Ang ZHAO
1
;
Li FAN
1
;
Bo MA
1
;
Xue-Jun SHANG
2
;
Qi ZHANG
1
Author Information
1. School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing, Jiangsu 211816, China.
2. epartment of Andrology, Jinling Hospital, Nanjing University School of Medicine / Nanjing General Hospital of Nanjing Military Region, Nanjing, Jiangsu 210002, China.
- Publication Type:Journal Article
- Keywords:
H₂O₂;
gas chromatography-mass spectrometry;
metabolomics;
mouse;
oxidative stress;
TM4 Sertoli cell
- MeSH:
Amino Acids;
metabolism;
Animals;
Apoptosis;
Cell Survival;
drug effects;
Dose-Response Relationship, Drug;
Energy Metabolism;
Gas Chromatography-Mass Spectrometry;
Glucose;
metabolism;
Hydrogen Peroxide;
pharmacology;
Male;
Metabolomics;
Mice;
Oxidative Stress;
Reactive Oxygen Species;
metabolism;
Sertoli Cells;
drug effects;
metabolism;
Time Factors
- From:
National Journal of Andrology
2017;23(6):497-502
- CountryChina
- Language:Chinese
-
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 H₂O₂. 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 H₂O₂ 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 H₂O₂ 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).
CONCLUSIONS: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.
