中华皮肤科杂志 ›› 2002, Vol. 35 ›› Issue (6): 429-431.

• 论著 • 上一篇    下一篇

氮氧化物对人角质形成细胞氧化性损伤的保护作用

孙凤兰1, 王国英2, 李其平3, 吴延芳2   

  1. 1. 青岛市海慈医院皮肤科 266033;
    2. 青岛大学医学院附属医院皮肤科 266033;
    3. 农业部动物产品监督监测中心
  • 收稿日期:2001-10-08 出版日期:2002-12-15 发布日期:2002-12-15
  • 基金资助:
    卫生部基金资助(96-2-329)

The Protective Effects of Nitroxides Against Oxidative Damage to Human Keratin ocytes

SUN Fenglan1, WANG Guoying2, LI Qiping3, WU Yanfang2   

  1. Department of Dermatology, Haici Hospital, Qingdao 266033, China
  • Received:2001-10-08 Online:2002-12-15 Published:2002-12-15

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摘要： 目的 观察氮氧化物对培养的人角质形成细胞氧化性损伤的保护作用,探讨其可能的作用机理.方法 在无血清、低Ca艹(0.1mmol/L)条件下培养人角质形成细胞,加入H2O2,造成角质形成细胞氧化性损伤,观察在氮氧化物存在及缺如的情况下细胞的存活率,细胞内还原型谷胱甘肽(GSH)含量和谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性变化.结果 ①H2O2可直接损伤角质形成细胞,H2O2作用浓度和细胞存活率呈高度负相关;②H2O2损伤可引起角质形成细胞内GSH含量降低,GSH-Px、SOD和CAT活性减弱;③预先加入氮氧化物2,2,6,6-四甲基哌啶(2,2,6,6-tetram-ethylpiperidine,TEMPO)或4-羟-2,2,6,6-四甲基哌啶(4-hydroxy-2,2,6,6-tetram-ethylpiperidine,TEMPOL)可抑制H2O2损伤引起的上述改变.结论 氮氧化物对培养的人角质形成细胞的氧化性损伤有保护作用,其机制与保护抗氧化酶活性,维持抗氧化系统平衡有关.

关键词: 氧化氮类, 角蛋白细胞, 抗氧化药

Abstract: Objective To study the protective effects of nitroxides against human keratinocytes oxidative damage induced by H2O2 and its possible mechanisms. Methods Normal human keratinocyte cultures obtained from foreskin were served as test-system. Human keratinocytes were cultured in human keratinocytes growth media (KGM) without serum and supplemented with 0.1 mmol/L Ca2+. Experiments were performed in culture when the cells grew to fuse. Oxidative damage was induced by adding H2O2 directly to the culture media at different concentrations in the present and absence of nitroxides [(2, 2, 6, 6-tetramethylpiperidine (TEMPO) a nd 4-hydroxy-2, 2, 6, 6-tetramethylpiperidine (TEMPOL)]. The cell viability was monitored and the intracellular level of reduced glutathione (GSH), the activities of glutathione-peroxidase(GSH-Px), superoxide dismutase (SOD), and catal ase were evaluated. Results ①H2O2 could cause damage to human keratinocytes directly. There was a significantly negative correlation between H2O2 concentration and cell viability. ②The level of GSH in keratinocytes lowered with treatment of H2O2. The activities of GSH-Px, SOD and catalase decreased. ③Pretreatment o f the cells with TEMPO and TEMPOL inhibited the damaging effects of H2O2 on cell viability and on cell antioxidant enzymatic systems. Conclusion The results of the study suggest that nitroxides may provide protection for cultured human kera tinocytes against H2O2-induced oxidative injury. It is proposed that the effects of nitroxides against cell oxidative damage be related to their protection of cellular enzymatic activities and maintaining cellular antioxidant systems.

Key words: Nitrogen oxides, Keratinocytes, Antioxidants