Protective and determining factors for the overall lipid peroxidation in ultraviolet A1-irradiated fibroblasts:: in vitro and in vivo investigations
Protective and determining factors for the overall lipid peroxidation in ultraviolet A1-irradiated fibroblasts:: in vitro and in vivo investigations
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DOI：
10.1046/j.1365-2133.2003.05457.x
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发表时间：
2003-08-01
影响因子：
10.3
通讯作者：
Scharffetter-Kochanek, K
中科院分区：
文献类型：
--
作者：
Dissemond, J;Schneider, LA;Scharffetter-Kochanek, K
脂质过氧化（LPO）是紫外线（UV）A导致光老化和促进皮肤癌的主要效应机制之一。目的探讨UVA 1诱导的人真皮成纤维细胞和小鼠真皮脂质过氧化损伤的相关因素，以及抗氧化酶在保护细胞免受脂质过氧化损伤中的作用。我们测量了丙二醛（MDA），一种LPO的特异性代谢示踪分子，以确定体外和体内由给定剂量的UVA 1产生的总LPO。通过使用铁螯合剂去铁胺（DFO）、羟基自由基清除剂二甲基亚砜（DMSO）和特异性过表达单一抗氧化酶的成纤维细胞，我们进一步间接评估了锰超氧化物歧化酶（MnSOD）的保护作用，过氧化氢酶和磷脂过氧化氢谷胱甘肽过氧化物酶以及不同活性氧的相对重要性和过渡态铁在不同UVA剂量诱导的LPO总量中的作用。体外MDA水平的依赖性增加，体外结果显示具有体内相关性。与DFO或DMSO孵育的成纤维细胞产生较低水平的MDA比对照组，如成纤维细胞过表达MnSOD，过氧化氢酶或PHGPx。结论细胞铁库和羟基自由基是最重要的决定因素后，产生的MDA总量的UVA 1剂量，和PHGPx过表达对LPO的保护作用最大。
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.