Quantitative measurements of oxidative stress in mouse skin induced by X-ray irradiation.
To find efficient methods to evaluate oxidative stress in mouse skin caused by X-ray irradiation, several markers and methodologies were examined. Hairless mice were irradiated with 50 Gy X-rays and skin homogenates or skin strips were prepared. Lipid peroxidation was measured using the skin homogenate as the level of thiobarbituric acid reactive substances. The level of lipid peroxidation increased with time after irradiation and was twice that of the control at 78 h. ESR spectra of skin strips showed a clear signal for the ascorbyl radical, which increased with time after irradiation in a manner similar to that of lipid peroxidation. To measure levels of glutathione (GSH) and its oxidized forms (GSSG) simultaneously, two HPLC methods, sample derivatization with 1-fluoro-2,4-dinitrobenzene and detection with a UV detector (method A) and no derivatization and detection with an electrochemical detector (method B), were compared and the latter was found to be better. No significant change was observed within 24 h after irradiation in the levels of GSH and GSSG measured by method B. The GSH/GSSG ratio may be a less sensitive parameter for the evaluation of acute oxidative stress caused by X-ray irradiation in the skin. Monitoring the ascorbyl radical seems to be a good way to evaluate oxidative stress in skin in vivo.
Chi
Cuiping
C
Redox Regulation Research Group, National Institute of Radiological Sciences, Chiba, Japan.
Tanaka
Ryoko
R
Okuda
Yohei
Y
Ikota
Nobuo
N
Yamamoto
Haruhiko
H
Urano
Shiro
S
Ozawa
Toshihiko
T
Anzai
Kazunori
K
D000818:Animals; D001205:Ascorbic Acid / Q000528:radiation effects; D001923:Brain Chemistry / Q000528:radiation effects; D004578:Electron Spin Resonance Spectroscopy; D005978:Glutathione / Q000737:chemistry; D005980:Glutathione Reductase / Q000737:chemistry; D007202:Indicators and Reagents; D015227:Lipid Peroxidation / Q000528:radiation effects; D008099:Liver / Q000737:chemistry / Q000528:radiation effects; D051379:Mice; D008812:Mice, Hairless; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000528:radiation effects*; D012867:Skin / Q000528:radiation effects*; D014965:X-Rays
D007202:Indicators and Reagents; D005980:Glutathione Reductase; D005978:Glutathione; D001205:Ascorbic Acid