The antioxidant system mediated by Nrf2 in C2C12 cells responding to HO stimulus under different oxygen concentration.
10.12047/j.cjap.5773.2019.067
- Author:
Wei-Xiu JI
1
;
Ying ZHANG
1
Author Information
1. Beijing Sport University, Beijing 100084.
- Publication Type:Journal Article
- MeSH:
Animals;
Antioxidants;
metabolism;
Cell Line;
Cell Survival;
Hydrogen Peroxide;
Mice;
Myoblasts;
enzymology;
NF-E2-Related Factor 2;
metabolism;
Oxidative Stress;
Oxygen;
Reactive Oxygen Species;
metabolism
- From:
Chinese Journal of Applied Physiology
2019;35(4):317-321
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To apply hypoxia of different oxygen concentration on C2C12 cells to study the changes of Nrf2 antioxidant system under HO.
METHODS:The perfect simulative effect time and concentration of HO were chosen. Cell vitality was tested after C2C12 cells cultured in 0.1 mmol/L, 0.25 mmol/L, 0.5 mmol/L, 0.75 mmol/L, 1 mmol/L and 2 mmol/L HO for 1 or 2 h respectively. The C2C12 cells were divided into different oxygen concentration group: 21%O, 12%O, 8%O, 5%O respectively. And then cells were treated with HO for 1 h, and collected for determination. Immunofluorescence of Nrf2 and the protein expression of Nrf2 were detected. The expressions of antioxidant enzymes superoxide dismutase 1 (SOD1), superoxide dismutase 2 (SOD2), catalase(CAT), NADPH quinine oxidoreductase-1 (NQO-1), glutathione peroxidase-1 (GPX-1), Heme oxygenase-1 (HO-1) mRNA and cellular ROS levels were tested by high quality fluorescence assay.
RESULTS:0.5 mmol/L HO for 1 h was selected as the conditions of HOstimulation. Compared with 21% O group, the expressions of Nrf2 mRNA and protein, antioxidant enzymes SOD1, SOD2, CAT, HO-1, NQO-1, GPX-1 mRNA were increased significantly (P＜0.05 or P＜0.01), and ROS level was lower (P＜0.01) in 12%O group cells; only the expression of GPX-1 mRNA was increased (P＜0.05) in 8%O group; the expressions of Nrf2 mRNA and protein expression, antioxidant enzymes SOD1, SOD2, NQO-1, GPX-1 mRNA were decreased significantly(P＜0.05 or P＜0.01), and ROS level was higher (P＜0.01) in 5%O group.
CONCLUSION:Hypoxia can affect the Nrf2 antioxidant system, and the different oxygen concentrations have different impact. In addition, 12% O for 12 h could promote the Nrf2 antioxidant system, and 5% extremely low oxygen may inhibit it.
