中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (2): 117-120.

• 论著 • 上一篇    下一篇

茶多酚EGCG及UVB对角质形成细胞水通道蛋白3表达及EGFR/ERK信号传导途径的影响

宋秀祖1,许文2,相文忠1,许爱娥3   

  1. 1. 杭州市第三人民医院皮肤科
    2.
    3. 安徽医科大学附属杭州市第三人民医院皮肤科
  • 收稿日期:2012-01-16 修回日期:2012-08-23 出版日期:2013-02-15 发布日期:2013-02-01
  • 通讯作者: 许爱娥 E-mail:xuaiehz@msn.com
  • 基金资助:
    a-MSH调控Th17/Treg平衡在黑素细胞免疫耐受中的作用研究;AQP3在皮肤甘油转运中的作用机制研究;皮肤屏障保护剂的前期开发研究

Effects of green tea polyphenol epigallocatechin?鄄3?鄄gallate and ultraviolet B on the expression of aquaporin 3 and EGFR/ERK signaling pathway in keratinocytes

  • Received:2012-01-16 Revised:2012-08-23 Online:2013-02-15 Published:2013-02-01

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摘要： 目的 探讨茶多酚表没食子儿茶素没食子酸酯（EGCG）及中波紫外线（UVB）对角质形成细胞水通道蛋白3（AQP3）的表达及信号传导途径的影响。 方法 20例健康人每日外涂不同浓度EGCG乳膏，2周后测定皮肤含水量及经皮水分丢失（TEWL）变化。对培养的人角质形成细胞分别予以10-7，10-6，10-5 mol/L的EGCG处理后予以UVB照射，或者用EGFR/ERK的磷酸化抑制剂处理后予以UVB照射，Western印迹检测AQP3蛋白表达变化以及EGFR/ERK信号传导途径变化。 结果 健康人皮肤予以不同浓度EGCG乳膏处理后，皮肤含水量显著增加，经皮水分丢失明显减少。UVB照射前予以10-7，10-6，10-5 mol/L EGCG处理，AQP3表达明显升高，分别为172.36 ± 12.42，320.66 ± 15.51，368.10 ± 11.39，与单纯UVB照射组（灰度值设为100.00）比较差异具有统计学意义（t值分别为12.16，26.75，38.62，P值均 < 0.05）。UVB照射前予以EGFR磷酸化抑制剂PD153035（1.0 μmol/L）和ERK磷酸化抑制剂U0126（10 μmol/L）处理后，AQP3表达也明显升高，分别为413.85 ± 25.27，268.85 ± 16.33，与单纯UVB照射组比较，差异具有统计学意义（t值分别为35.16，19.25，P值均 < 0.05）。UVB照射可以激活角质形成细胞EGFR/ERK信号传导途径，预先予以EGCG处理可以显著抑制EGFR/ERK的磷酸化。结论 EGCG可以增强皮肤屏障功能。UVB照射可以下调角质形成细胞AQP3表达，而EGCG处理可以上调AQP3表达，其机制可能与抑制UVB照射诱导的EGFR/ERK活化有关。

关键词: 皮肤屏障, 水通道蛋白3, 中波紫外线, 表没食子儿茶素没食子酸酯

Abstract: SONG Xiu-zu, XU Wen, XIANG Wen-zhong, XU Ai-e. Department of Dermatology, Third People's Hospital of Hangzhou, Hangzhou 310009, China Correponding author：XU Ai-e, Email: xuaiehz@msn.com 【Abstract】 Objective To evaluate the effect of green tea polyphenol epigallocatechin-3-gallate （EGCG） and ultraviolet B （UVB） on the expression of aquaporin 3 and epidermal growth factor receptor （EGFR）/extracellular signal-regulated protein kinase （ERK） signaling pathway in keratinocytes. Methods Twenty healthy human subjects were enrolled in this study. Both legs of each subjects were separated into 4 areas to remain untreated （control area）, be topically treated with 3% and 1% EGCG cream and the vehicle of EGCG cream respectively once a day for 2 weeks followed by the measurement of skin moisture content and trans-epidermal water loss （TEWL）. Cultured keratinocytes were classified into various groups to be irradiated with different doses （10, 20 and 30 mJ/cm2） of UVB, or be pretreated with different concentrations of EGCG （10－7， 10－6，10－5 mol/L） or EGFR/ERK phosphorylation inhibitors for 1 hour followed by irradiation with UVB of 30 mJ/cm2. After various durations of additional culture, Western blot was conducted to quantify the expression of AQP3 and phosphorylated-EGFR （p-EGFR） and -ERK （p-ERK） of keratinocytes. Data were processed by SPSS 10.0 software, and statistical analysis was carried out by t test. Results Skin moisture content was significantly increased, while TEWL was decreased in healthy skin after treatment with 1% and 3% EGCG cream compared with vehicle-treated skin areas and untreated skin areas. Increased AQP3 expression was observed in keratinocytes pretreated with EGCG of 10－7，10－6，10－5 mol/L （172.36 ± 12.42，320.66 ± 15.51 and 368.10 ± 11.39 vs. 100.00, t = 12.16， 26.75 and 38.62 respectively, all P < 0.05） and in those pretreated with the EGFR inhibitor PD153035 of 1.0 μmol/L and ERK inhibitor U0126 of 10 μmol/L（413.85 ± 25.27 and 268.85 ± 16.33 vs. 100.00, t = 35.16, 19.25 respectively, both P < 0.05）compared with those irradiated with UVB of 30 mJ/cm2 alone. UVB irradiation stimulated the phosphorylation of EGFR/ERK in keratinocytes, and the stimulation was markedly inhibited by the pre-treatment with EGCG of 10－7， 10－6 and 10－5 mol/L （all P < 0.05）. Conclusions EGCG can enhance skin barrier function. AQP3 expression is down-regulated by UVB irradiation in keratinoctyes, while EGCG can inhibit the downregulation likely by suppressing the UVB-induced activation of EGFR and ERK. 【Key words】 Skin barrier； Aquaporin 3； Ultraviolet B；EGCG

Key words: aquaporin3, Ultraviolet B, EGCG