中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (2): 77-79.

• 论著 • 上一篇    下一篇

核因子-κB反义寡核苷酸抑制紫外线诱导的HaCaT细胞反义分泌白介素6

夏济平, 宋秀祖, 毕志刚   

  1. 南京医科大学第一附属医院皮肤科 210029
  • 收稿日期:2004-03-05 出版日期:2005-02-15 发布日期:2005-02-15
  • 通讯作者: 毕志刚,E-mail:eltonbi@21cn.com E-mail:eltonbi@21cn.com
  • 基金资助:
    国家自然基金资助项目(30271195)

Effects of Antisense NF-κB p65 Oligodeoxynucleotides on UV-induced IL-6 Secretion of Keratinocytes

XIA Ji-ping, SONG Xiu-zu, BI Zhi-gang   

  1. Department of Dermatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
  • Received:2004-03-05 Online:2005-02-15 Published:2005-02-15

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摘要： 目的 探讨核因子-κB(NF-κB)反义寡核苷酸转染体外培养的HaCaT细胞反义株HaCaT后,对紫外线(UV)诱导的HaCaT细胞反义分泌炎症因子白介素6(IL-6)的抑制作用。方法 蛋白质印迹方法测定不同剂量中波紫外线(UVB)辐射HaCaT细胞反义后NF-κBp65的变化;逆转录-聚合酶链反应测定NF-κBp65反义寡核苷酸转染后p65m RNA表达的变化;酶联免疫吸附测定法测定NF-κBp65反义寡核苷酸转染后紫外线辐射的HaCaT细胞反义分泌IL-6水平。结果 10、20、30mJ/cm2 UVB辐照HaCaT细胞反义后均可显著促进NF-κBp65表达(P<0.05),不同浓度的NF-κBp65反义寡核苷酸转染后对30mJ/cm2 UVB诱导的p65mRNA表达和IL-6分泌均有显著抑制作用(P<0.05)。结论 脂质体介导的NF-κBp65反义寡核苷酸转染HaCaT细胞反义,可以抑制UV辐射诱导的HaCaT细胞反义产生IL-6,表明UV诱导HaCaT细胞反义产生IL-6可能是通过UV激活NF-κBp65的表达产生的。

关键词: 紫外线, 角蛋白细胞, NF-κB, 寡核苷酸类, 反义

Abstract: Objective To explore the effects of antisense NF-κB p65 oligodeoxynucleotides on UV-induced IL-6 secretion of keratinocytes. Methods NF-κB p65 oligodeoxynucleotides were transfected to a keratinocyte cell line, HaCaT cells, mediated by lipofectamine. The NF-κB p65 protein in the cells was measured by Western blot assay, the mRNA level of NF-κB p65 was detected by RT-PCR, and UV-induced IL-6 level was determined by ELISA pre-and post-transfection and/or UVB irradiation. Results The NF-κB p65 protein expression was significantly increased in UVB (10, 20, 30 mJ/cm2) irradiation groups as compared with that of the control groups (P<0.05). The p65 mRNA and UVB-induced IL-6 were inhibited after the keratinocytes were transfected with p65 antisense oligodeoxynucleotides at the concentrations of 2, 4 and 8 μg/mL (P<0.05). Conclusions UV-induced IL-6 secretion of keratinocytes might be mediated by NF-κB, and could be inhibited by NF-κB p65 antisense oligodeoxynucleotides.

Key words: Ultraviolet rays, Keratinocytes, NF-kappa B, Oligonucleotides, antisense