Inverse regulation of the nuclear factor-κB binding to the p53 and interleukin-8 κB response elements in lesional psoriatic skin
Inverse regulation of the nuclear factor-κB binding to the p53 and interleukin-8 κB response elements in lesional psoriatic skin
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DOI：
10.1111/j.0022-202x.2005.23749.x
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发表时间：
2005-06-01
影响因子：
6.5
通讯作者：
Iversen, L
中科院分区：
文献类型：
--
作者：
Johansen, C;Flindt, E;Iversen, L
核因子-κ B（NF-κ B）是一种可诱导的核转录因子，调节一系列细胞过程。因此，NF-κ B B的DNA结合活性失衡可能是银屑病病理生理机制的一部分。本研究的目的是使用三种不同的kappa B位点测定银屑病皮肤中NF-κ B B DNA结合活性，并确定抗银屑病药物卡泊三醇如何调节DNA结合活性。通过电泳迁移率变动分析，我们证明，与非皮损型银屑病皮肤相比，皮损型银屑病皮肤中NF-κ B B DNA与p53 κ B位点的结合减少，而NF-κ B B DNA与白细胞介素-8（IL-8）κ B位点的结合增加。未观察到NF-κ B DNA与主要组织相容性复合体I类κ B位点结合的调节。通过定量RT-PCR测定，与未受累皮肤相比，受累银屑病皮肤中p53表达的类似降低和IL-8表达的增加证实了这些变化。NF-κ B DNA结合活性的改变既不伴随抑制剂κ B（I κ B）激酶、IKK α、IKK β和IKK γ表达的任何变化，也不伴随NF-κ B抑制剂蛋白、I κ B α和I κ B β表达的任何变化。免疫荧光分析显示，p65被隔离在角质形成细胞的细胞质中，而p50表现出细胞质以及核定位。有趣的是，p50和p65的这种分布在皮损和非皮损银屑病皮肤中是相似的。卡泊三醇局部应用于银屑病皮损4 d导致NF-κ B与p53 κ B位点结合增加，NF-κ B与IL-8 κ B位点结合减少。综上所述，我们的数据表明，NF-κ B DNA结合活性在银屑病皮肤中以特定的方式受到调节，这取决于所研究的κ B位点，并且银屑病皮肤的局部治疗使在皮损银屑病皮肤中观察到的异常NF-κ B结合活性正常化。
Nuclear factor-kappa B (NF-kappa B) is an inducible nuclear transcription factor regulating a range of cellular processes. An imbalance of the DNA binding activity of NF-kappa B may, therefore, be part of the pathophysiological mechanisms in psoriasis. The purpose of this study was to determine the NF-kappa B DNA binding activity in psoriatic skin using three different kappa B sites and to determine how DNA binding activity was modulated by the anti-psoriatic drug calcipotriol. By electrophoretic mobility shift assay, we demonstrated that the NF-kappa B DNA binding to the p53 kappa B site was decreased, whereas the NF-kappa B DNA binding to the interleukin-8 (IL-8) kappa B site was increased in lesional psoriatic skin compared with non-lesional psoriatic skin. No regulation was seen on the NF-kappa B DNA binding to the major histocompatibility complex class I kappa B site. These changes were paralleled by a similar decrease in p53 expression and an increase in IL-8 expression in involved psoriatic skin compared with uninvolved skin as determined by quantitative RT-PCR. The alteration in NF-kappa B DNA binding activity was neither accompanied by any change in the expression of the inhibitor kappa B (I kappa B) kinases, IKK alpha, IKK beta, and IKK gamma nor in the expression of the NF-kappa B inhibitor proteins, I kappa B alpha and I kappa B beta. Immunofluorescence analysis revealed that p65 was sequestered in the cytoplasm of keratinocytes, whereas p50 exhibited a cytoplasmic as well as a nuclear localization. Interestingly, this distribution of p50 and p65 was similar in lesional and non-lesional psoriatic skin. Topical application of calcipotriol to lesional psoriatic skin for 4 d resulted in increased NF-kappa B binding to the p53 kappa B site and decreased NF-kappa B binding to the IL-8 kappa B site. Taken together, our data demonstrate that the NF-kappa B DNA binding activity is regulated in a specific manner in psoriatic skin depending on the kappa B sites investigated, and that topical treatment of psoriatic skin normalizes the abnormal NF-kappa B binding activity seen in lesional psoriatic skin.