中华皮肤科杂志

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中波紫外线辐射对人表皮角质形成细胞核因子κB转录活性的影响

王平1, 毕志刚2, 许爱娥1, 崔毓桂3   

  1. 1. 杭州市第三人民医院皮肤科, 310009;
    2. 南京医科大学第一附属医院皮肤科;
    3. 南京医科大学第一附属医院中心实验室
  • 收稿日期:2005-11-21 出版日期:2006-04-15 发布日期:2006-04-15
  • 通讯作者: 毕志刚,email:eltonbi@21cn.com E-mail:eltonbi@21cn.com
  • 基金资助:
    国家自然科学基金(30271195)

Effect of ultraviolet B irradiation on the transcriptional activity of nuclear factor κB in human epidermal keratinocytes

WANG Ping1, BI Zhi-gang2, XU Ai-e1, CUI Yu-gui3   

  1. Department of Dermatology, Third Hospital of Hangzhou, Hangzhou 310009, China
  • Received:2005-11-21 Online:2006-04-15 Published:2006-04-15

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摘要： 目的 探讨中波紫外线(UVB)辐射对表皮角质形成细胞核因子κB(NF-κB)转录活性的影响。方法 正常人表皮角质形成细胞取自健康儿童包皮,并培养于37℃、5%CO2的培养箱中;以20、60和120 mJ/cm2 UVB辐射培养细胞;分别提取UVB辐射0、2、24和48 h后角质形成细胞的全细胞蛋白、核蛋白和胞浆蛋白;免疫印迹方法检测分析全细胞蛋白和核蛋白的NF-κB/p65以及胞浆蛋白NF-κB抑制物(IκB-a)的动态变化;电泳迁移率变更分析(EMSA)方法检测分析NF-κB的转录活性。结果 UVB辐射可显著促进角质形成细胞总蛋白和核蛋白中NF-κB的蛋白表达(P<0.05),且表达量与辐射剂量成正比,UVB辐射后2 h即可有NF-κB的蛋白表达水平的升高。24 h后达高峰并持续高水平表达至48 h:不同剂量UVB辐射角质形成细胞后的不同时间,均可促进NF-κB转录活性:UVB辐射可显著抑制角质形成细胞胞浆IκB-a蛋白的表达(P<0.05)。结论 UVB辐射可显著促进角质形成细胞NF-κB的转录活性。

关键词: 紫外线, 角蛋白细胞, NF-κB

Abstract: Objective To study the effect of ultraviolet B (UVB) irradiation on the transcriptional activity of nuclear factor KB(NF-κB) in human epidermal keratinocytes.Methods Normal human epidermal keratinocytes were isolated from healthy children foreskin and cultured at 37℃ in 5% CO2.Cultured keratinocytes were irradiated with UVB at the dose of 20,60 and 120 mJ/cm2,respectively.The whole cell, nuclear and cytoplasmic protein of keratinocytes was extracted at 0,2,24,and 48 hours after the irradiation. The whole cell and nuclear NF-κB/p65 protein as well as cytoplasmic NF-κB Inhibitor(IκB-a) protein were detected by Western blotting.The transcriptional activity of NF-κB was analyzed by electrophoretic mobility shift assay (EMSA).Results UVB irradiation significantly increased (P<0.05) the protein expression of NF-κB in both the whole cell and the nucleus of keratinocytes in a dose-dependent manner;the increase began at 2 h,peaked at 24 h,and maintained with high expression until 48 h after the irradiation. The transcriptional activity of NF-κB was improved at different time points after UVB irradiation of different doses,while the expression of Iκd3-a was significantly decreased (P<0.05) by the irradiation.Conclusion UVB irradiation can improve the transcriptional activity of keratinocytes,likely by the degradation of IκB-a.

Key words: UVB, Keratinocytes, NF-kappa B