Effect of hydrogen-rich water on oxidative stress of in vitro orbital fibroblasts of patients with thyroid associated ophthalmopathy
10.16781/j.0258-879x.2018.05.0547
- Author:
Xin-Xin CHEN
1
Author Information
1. Department of Ophthalmology, Changzheng Hospital, Navy Medical University (Second Military Medical University)
- Publication Type:Journal Article
- Keywords:
Dexamethasone;
Hydrogen-rich water;
Orbital fibroblasts;
Oxidative stress;
Thyroid associated ophthalmopathy
- From:
Academic Journal of Second Military Medical University
2018;39(5):547-551
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the anti-oxidation stress effect of hydrogen-rich water (HW) on thyroid associated ophthalmopathy (TAO). Methods The orbital fibroblasts were derived from orbital adipose connective tissue of TAO patients and cultured in vitro. The cells were treated with different concentrations of hydrogen peroxide (H2O2) for 18 h, and the proliferation ability was detected by CCK-8 method to determine the appropriate H2O2 concentration. The cells were divided into four groups: blank group (normal culture), H2O2 group (treating cells with H2O2 for 18 h), HW+H2O2 group (culturing cells using culture media containing HW for 72 h in combination with H2O2 for 18 h), dexamethasone + H2O2 group (treating cells using dexamethasone 1 μmol/L for 72 h in combination with H2O2 for 18 h). After culturing for 72 h in each group, the fluorescence intensity of reactive oxygen species (ROS) was measured by flow cytometry, and the contents of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected by ELISA. Results We successfully cultured orbital fibroblasts derived from orbital adipose connective tissues of TAO patients. The higher concentration of H2O2, the greater inhibition effect on the proliferation ability of the orbital fibroblasts from TAO patients, and we finally chose 100 μmol/L H2O2. After 72 h of cell culture, the contents of MDA, SOD, and GSHPx and the fluorescence intensity of ROS were (1.63±0.29), (5.06±0.24), (3.94±0.29), and (2.34±0.24) nmol/mL, (10.51±0.32), (2.41±0.23), (5.58±0.29), and (7.98±0.15) U/mL, (107.79±1.06), (21.07±0.92), (49.19±6.75), and (76.33±4.70) U/mL and 18 275.82±521.05, 92 524.81±2 097.01, 54 460.87±572.64, and 35 961.37±540.61 in the blank, H2O2, HW+H2O2 and dexamethasone+H2O2 groups, respectively. Statistic analysis results showed that both HW and dexamethasone could significantly inhibit oxidative stress induced by H2O2 in orbital fibroblasts (all P<0.01), and the inhibitory effects of dexamethasone were significantly more obvious than those of HW (all P<0.01). Conclusio HW may be a treatment option for TAO through anti-oxidant stress.
