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Glutathione modulates the level of free radicals produced in UVA irradiated cells.

Tobi, Simon E. and Paul, Nigel and McMillan, Trevor J. (2001) Glutathione modulates the level of free radicals produced in UVA irradiated cells. Journal of Photochemistry and Photobiology B: Biology, 57 (2-3). pp. 102-112. ISSN 1011-1344

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We have developed an assay to detect reactive oxygen species (ROS) generated by UVA radiation utilising chemical probes which become fluorescent upon oxidation. Using a human bladder carcinoma cell line (MGH-U1) and spontaneously immortalised keratinocytes (HaCaT), we have shown a UVA (narrow band 365±5 nm) dose-dependent increase in fluorescence by flow cytometry following loading of the cells with either dihydrorhodamine 123 (DHR) or 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA). The UVA response of both DHR and DCFH was enhanced by elevation of intracellular levels of the photosensitiser protoporphyrin IX by incubation for 2.5 h with 5-aminolaevulinic acid. Depletion of the antioxidant glutathione (GSH) using the inhibitor -buthionine-sulphoximine (BSO), resulted in an increase in the UVA-induced fluorescence of DCF but not of rhodamine 123. Conversely, raising intracellular GSH levels with N-acetyl cysteine (NAC) had relatively little protective effect in terms of degree of induced fluorescence.

Item Type: Journal Article
Journal or Publication Title: Journal of Photochemistry and Photobiology B: Biology
Uncontrolled Keywords: UVA ; Reactive oxygen species ; Glutathione ; 2′ ; 7′-Dichlorodihydrofluorescein diacetate
Subjects: ?? qh301 ??
Departments: Faculty of Science and Technology > Lancaster Environment Centre
VC's Office
ID Code: 8726
Deposited By: Professor Trevor McMillan
Deposited On: 09 May 2008 10:15
Refereed?: Yes
Published?: Published
Last Modified: 10 Apr 2018 12:12
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