Protective and determining factors for the overall lipid peroxidation in ultraviolet A1-irradiated fibroblasts: in vitro and in vivo investigations

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Abstract:

Summary 

Background Lipid peroxidation (LPO) is one major effector mechanism by which ultraviolet (UV) A contributes to photoageing and the promotion of skin cancer. It is a fingerprint of photo-oxidative stress within the skin, and is initiated by several pathways, with different reactive oxygen species (ROS) and iron ions being involved.

Objectives To elucidate factors involved in UVA1-induced LPO in human dermal fibroblasts and mouse dermis, and the role of antioxidant enzymes in protecting cells against LPO.

Methods Using a highly sensitive high-performance liquid chromatography procedure, we measured malondialdehyde (MDA), a specific metabolic tracer molecule for LPO, to determine the overall LPO produced by a given UVA1 dose in vitro and in vivo. By using the iron chelator desferrioxamine (DFO), the hydroxyl radical scavenger dimethylsulphoxide (DMSO) and fibroblasts that specifically overexpress single antioxidant enzymes, we further indirectly assessed the protective effect of manganese superoxide dismutase (MnSOD), catalase and phospholipid hydroperoxide glutathione peroxidase (PHGPx) as well as the relative importance of different ROS and the role of transitional iron for the total amount of LPO induced by a distinct UVA dose.

Results UVA1 irradiation resulted in a time- and dose-dependent increase in MDA levels in vitro, and the in vitro results were shown to have in vivo relevance. Fibroblasts incubated with DFO or DMSO produced lower levels of MDA than controls, as did fibroblasts overexpressing MnSOD, catalase or PHGPx.

Conclusions The cellular iron pool and hydroxyl radicals were the most important determining factors for the total amount of MDA produced after a given UVA1 dose, and PHGPx overexpression had the greatest protective effect against LPO.

Keywords: antioxidant enzymes; desferrioxamine; dimethylsulphoxide; fibroblast; lipid peroxidation; malondialdehyde; oxidative stress; ultraviolet A

Document Type: Research Article

DOI: http://dx.doi.org/10.1046/j.1365-2133.2003.05457.x

Affiliations: 1: Department of Dermatology, University of Essen, Hufelandstrasse 55, D-45122 Essen, Germany 2: Department of Dermatology, University of Ulm, Maienweg 12, D-89081 Ulm, Germany 3: Department of Physiological Chemistry I, Heinrich-Heine University Düsseldorf, PO Box 101007, D-40001 Düsseldorf, Germany

Publication date: August 1, 2003

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