Effects of ultraviolet B irradiation on the production of regulated upon activation normal T-cell expressed and secreted protein in cultured human epidermal keratinocytes
Effects of ultraviolet B irradiation on the production of regulated upon activation normal T-cell expressed and secreted protein in cultured human epidermal keratinocytes
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DOI：
10.1007/s00403-005-0620-6
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发表时间：
2006-02-01
影响因子：
3
通讯作者：
Fujiwara, S
中科院分区：
文献类型：
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作者：
Arakawa, S;Hatano, Y;Fujiwara, S
紫外线B （UVB）照射对皮肤炎症反应的调节作用是众所周知的，但其机制尚不清楚。正常t细胞表达和分泌蛋白（RANTES）是表皮角质形成细胞产生的趋化因子之一，经活化后的正常t细胞表达和分泌蛋白（RANTES）可能在皮肤炎症性疾病，如特应性皮炎和寻常性牛皮癣的发病机制中起重要作用。本研究旨在确定UVB照射是否会影响培养的正常人表皮角质形成细胞在炎症因子刺激下诱导的RANTES的产生。我们分别通过半定量反转录和聚合酶链反应以及酶联免疫吸附试验（ELISA）测量了培养角质形成细胞中RANTES基因的转录水平和培养上清中RANTES本身的转录水平。在没有肿瘤坏死因子α (tnf - α)和/或干扰素γ (ifn - γ)事先刺激细胞的情况下，转录本和RANTES本身都没有被检测到，RANTES的产生也没有被单独的UVB （100 J/m(2)）照射诱导。在培养基中加入tnf - α和ifn - γ之前，用UVB照射细胞，然后在12、24和36小时后收获细胞和培养上清。在辐照和未辐照的细胞中，RANTES mRNA在12 h时首次检测到，随后水平升高。在24 h时检测到RANTES本身，在36 h时检测到较高水平。在检测的所有时间点，UVB照射抑制了RANTES mRNA和蛋白质本身的产生。这些结果表明，UVB照射对皮肤炎症的调节作用可能与表皮角质形成细胞抑制RANTES的产生有关。
The modulatory effects of ultraviolet B (UVB) irradiation on cutaneous inflammatory responses are well known but their mechanism remains obscure. It has been proposed that regulated upon activation normal T-cell expressed and secreted protein (RANTES), which is one of the chemokines produced by epidermal keratinocytes, might play an important role in the pathogenesis of cutaneous inflammatory disorders, such as atopic dermatitis and psoriasis vulgaris. This study was designed to determine whether UVB irradiation could affect the production of RANTES that is induced in cultured normal human epidermal keratinocytes upon stimulation by inflammatory cytokines. We measured levels of the transcript of the gene for RANTES in cultured keratinocytes and of RANTES itself in culture supernatants by semiquantitative reverse transcription and the polymerase chain reaction and by an enzyme-linked immunosorbant assay (ELISA), respectively. Neither the transcript nor RANTES itself was detected without prior stimulation of cells by tumor necrosis factor alpha (TNF-alpha) and/or interferon gamma (IFN-gamma) and production of RANTES was not induced by UVB (100 J/m(2)) irradiation alone. Cells were irradiated with UVB just before addition of TNF-alpha and IFN-gamma to the medium and then cells and culture supernatants were harvested 12, 24, and 36 h later. In both irradiated and non-irradiated cells, RANTES mRNA was first detected at 12 h and the level increased subsequently. RANTES itself was detected at 24 h, with a higher level at 36 h. At all time points examined, UVB irradiation inhibited the production of RANTES mRNA and of the protein itself. These results suggest that suppression of the production of RANTES by epidermal keratinocytes might be involved in the modulatory effects of UVB irradiation on cutaneous inflammation.