CAJ | 학술논문

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/LEGCG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.
목적 탐토다다분표몰식자인다소몰식자산지(EGCG)급중파자외선(UVB)대각질형성세포수통도단백3 (AQP3)적표체급신호전도도경적영향.방법 20례건강인매일외도불동농도EGCG유고,2주후측정피부함수량급경피수분주실(TEWL)변화.대배양적인각질형성세포분별여이10-7,10-6,10-5 mol/L적EGCG처리후여이UVB조사,혹자용EGFR/ERK적린산화억제제처리후여이UVB조사,Western인적검측AQP3단백표체변화이급EGFR/ERK신호전도도경변화.결과 건강인피부여이불동농도EGCG유고처리후,피부함수량현저증가,경피수분주실명현감소.UVB조사전여이10-7,10-6,10-5 mol/LEGCG처리,AQP3표체명현승고,분별위172.36±12.42,320.66±15.51,368.10±11.39,여단순UVB조사조(회도치설위100.00)비교차이구유통계학의의(t치분별위12.16,26.75,38.62,P치균＜0.05).UVB조사전여이EGFR린산화억제제PD153035(1.0 μmol/L)화ERK린산화억제제U0126(10 μmol/L)처리후,AQP3표체야명현승고,분별위413.85±25.27,268.85±16.33,여단순UVB조사조비교,차이구유통계학의의(t치분별위35.16,19.25,P치균＜0.05).UVB조사가이격활각질형성세포EGFR/ERK신호전도도경,예선여이EGCG처리가이현저억제EGFR/ERK적린산화.결론 EGCG가이증강피부병장공능.UVB조사가이하조각질형성세포AQP3표체,이EGCG처리가이상조AQP3표체,기궤제가능여억제UVB조사유도적EGFR/ERK활화유관.
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 transepidermal 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.