中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (7): 413-415.

• 论著 • 上一篇    下一篇

紫外线A1对真皮成纤维细胞的损伤及Tempol保护作用的研究

严淑贤1, 胡跃1, 邓辉1, 洪新宇2, 徐昱1, 廖康煌1   

  1. 1. 复旦大学附属华山医院皮肤科 上海 200040;
    2. 复旦大学公共卫生学院皮肤生理毒理研究室 上海 200040
  • 收稿日期:2004-07-19 发布日期:2005-07-15
  • 通讯作者: 胡跃, E-mail:huyue1104@hotmail.com E-mail:huyue1104@hotmail.com

Protective Effects of Tempol on Ultraviolet-AI Irradiated Human Dermal Fibroblasts

YAN Shu xian1, HU Yue1, DENG Hui1, HONG Xin-yu2, XU Yu1, LIAO Kang-huang1   

  1. Department of Dermatology, Huashan Hospital of Fudan University, Shanghai 200040, China
  • Received:2004-07-19 Published:2005-07-15

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摘要： 目的 研究紫外线Al(UVAl,340nm-400nm)对人真皮成纤维细胞氧化性损伤、对基质金属蛋白酶(MMP)和胶原蛋白表达的影响,观察氮氧化物Tempol对这些改变的保护作用。方法 体外原代培养的人真皮成纤维细胞接受UVAl照射,同时加入或不加Tempol,以生化方法检测细胞超氧化物歧化酶(SOD)活性和脂质过氧化物丙二醛(MDA)含量,以半定量逆转录-聚合酶链反应及酶连免疫吸附试验测定MMP-1,MMP-3,Ⅰ型及Ⅲ型胶原mRNA或蛋白表达水平。结果 15J/cm2UVAl照射可显著抑制细胞SOD活性,提高MDA含量,促进MMP-1,MMP-3mRNA表达,同时显著抑制Ⅰ型和Ⅲ型胶原蛋白表达(P<0.05)。照射同时加入Tempol,在一定浓度范围,可显著抑制UVAl引起的上述改变(P<0.05)。结论 Tempol在体外对UVAl引起的氧化性损伤以及真皮细胞外基质成分代谢紊乱有保护作用,因而有可能成为防治光老化的一种成分。

关键词: 紫外线, 活性氧, 成纤维细胞, 超氧化物歧化酶, 丙二醛

Abstract: Objective To determine the effects of Tempol,one of the nitroxides,in the presence of ultraviolet-AI (UVA1,340 nm-400 nm) on superoxide enzyme (SOD) activity,lipid peroxidation,and expression of collagen Ⅰ,collagen Ⅲ and matrix metalloproteinases (MMP)-1,MMP-3 in human dermal fi broblasts in vitro.Methods Fibroblasts were irradiated by a single exposure to UVA1 and at the same time incubated with or without Tempol,and detected 24 h later.SOD activity and lipid peroxidation,as shown by accumulation of malondialdehyde (MDA),were detected by biochemical assay.Expression of collagen Ⅰ,collagen Ⅲ (protein levels) and MMP-1,MMP-3 (mRNA level) was examined by ELISA and semi-quantitative RT-PCR,respectively.Results A dose of 15 J/cm2 UVA1 significantly inhibited SOD activity and collagen Ⅰ ,collagen Ⅲ protein levels,increased MDA level and stimulated MMP-1,MMP-3 mRNA expression (P<0.05).Tempol could reverse these effects caused by UVA1 to some degree or completely,and with proper concentration,the results were statistically significant (P<0.05).Conclusions Tempol exerts photoprotective activities against UVA1 irradiation in vitro.With antioxidant ability,Tempol may normalize extracellular matrix metabolism in dermis and could be used as an anti-photoageing agent.

Key words: Ultraviolet rays, Reactive oxygen species, Fibroblasts, Superoxide dismutase, Malon dialdehyde