甘草次酸抑制表皮细胞生长因子对HaCaT细胞促增殖作用的机制研究
감초차산억제표피세포생장인자대HaCaT세포촉증식작용적궤제연구
Mechanisms underlying the inhibitory effect of glycyrrhetinic acid on epidermal growth factor-induced proliferation of HaCaT cells
  • 万方数据
    中华皮肤科杂志 중화피부과잡지
    Chinese Journal of Dermatology
    2013年 4期 278-281 ,共4页

    解凡%曹毅%刘改荣%杨晓红%代群%陈微

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    目的 探讨甘草次酸对表皮细胞生长因子(EGF)诱导的HaCaT细胞增殖的影响,并对其可能机制进行相关研究.方法 MTT法检测不同浓度EGF(0、1、5、10、25、50、100μg/L)以及不同浓度甘草次酸(0、0.1、1、10、25、50、100 μmol/L)对HaCaT细胞增殖的影响,25 μmol/L甘草次酸、10 μmol/L MEK1/2抑制剂(U0126)对25 μg/L EGF促HaCaT细胞增殖的抑制作用.蛋白免疫印迹法检测25 μg/L EGF对ERK1/2磷酸化的促进作用,25 μmol/L甘草次酸、10 μmol/L U0126对ERK1/2磷酸化的抑制作用,25 μg/L EGF、25 μmol/L甘草次酸、10 μmol/L U0126分别或同时作用下HaCaT细胞增殖细胞核抗原(PCNA)、Notch-1蛋白的表达情况.使用SPSS17.0统计软件对实验数据进行t检验、方差分析和相关分析.结果 EGF在0～100 μg/L范围内促进HaCaT细胞增殖(r=0.798,P＜0.05),在0～50g/L范围内与HaCaT细胞增殖呈线性相关(r=0.859,P＜0.05).甘草次酸在10～100 μmol/L范围内浓度依赖性地抑制HaCaT细胞增殖(r=-0.945,P＜ 0.01)；25 μmol/L甘草次酸明显抑制25 μg/L EGF对HaCaT细胞的促增殖作用以及对ERK1/2的促磷酸化作用.同时,25μmol/L甘草次酸、10 μmol/L U0126均抑制25 μg/L EGF对HaCaT细胞PCNA、Notch-1蛋白的促表达作用.结论 甘草次酸可抑制EGF对HaCaT细胞的促增殖作用,其机制可能与甘草次酸抑制EGF对HaCaT细胞ERK1/2信号通路的激活有关.
    목적 탐토감초차산대표피세포생장인자(EGF)유도적HaCaT세포증식적영향,병대기가능궤제진행상관연구.방법 MTT법검측불동농도EGF(0、1、5、10、25、50、100μg/L)이급불동농도감초차산(0、0.1、1、10、25、50、100 μmol/L)대HaCaT세포증식적영향,25 μmol/L감초차산、10 μmol/L MEK1/2억제제(U0126)대25 μg/L EGF촉HaCaT세포증식적억제작용.단백면역인적법검측25 μg/L EGF대ERK1/2린산화적촉진작용,25 μmol/L감초차산、10 μmol/L U0126대ERK1/2린산화적억제작용,25 μg/L EGF、25 μmol/L감초차산、10 μmol/L U0126분별혹동시작용하HaCaT세포증식세포핵항원(PCNA)、Notch-1단백적표체정황.사용SPSS17.0통계연건대실험수거진행t검험、방차분석화상관분석.결과 EGF재0～100 μg/L범위내촉진HaCaT세포증식(r=0.798,P＜0.05),재0～50g/L범위내여HaCaT세포증식정선성상관(r=0.859,P＜0.05).감초차산재10～100 μmol/L범위내농도의뢰성지억제HaCaT세포증식(r=-0.945,P＜ 0.01)；25 μmol/L감초차산명현억제25 μg/L EGF대HaCaT세포적촉증식작용이급대ERK1/2적촉린산화작용.동시,25μmol/L감초차산、10 μmol/L U0126균억제25 μg/L EGF대HaCaT세포PCNA、Notch-1단백적촉표체작용.결론 감초차산가억제EGF대HaCaT세포적촉증식작용,기궤제가능여감초차산억제EGF대HaCaT세포ERK1/2신호통로적격활유관.
    Objective To estimate the effect of glycyrrhetinic acid on epidermal growth factor (EGF)-induced proliferation of HaCaT cells,and to investigate its possible mechanism.Methods Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the proliferation of HaCaT cells treated with different concentrations of EGF (0,1,5,10,25,50,100 μg/L) and glycyrrhetinic acid (0,0.1,1.0,10,25,50,100μmol/L) alone,or the combination of 25 μg/L EGF with 25 μ mol/L glycyrrhetinic acid or 10 μ mol/L U0126 (an inhibitor of MEK1/2).Western blot was carried out to measure the protein expression of proliferating cell nuclear antigen (PCNA),Notch-1,ERK 1/2 and phosphorylated ERK 1/2 in HaCaT cells treated with 25 μg/L EGF,10 μmol/L U0126,25μmol/L glycyrrhetinic acid alone or in combination.Data were statistically analyzed by using t test,analysis of variance and correlation analysis with SPSS 17.0 software.Results EGF of 0-100 μg/L promoted the proliferation of HaCaT cells in a dose-dependent manner (r =0.798,P ＜ 0.05),and there was a linear correlation between the effect and concentration within the concentration range 0-50 μg/L (r =0.859,P ＜ 0.05).However,glycyrrhetinic acid of 10-100 μmol/L inhibited the proliferation of HaCaT cells in a dose-dependent manner (r =-0.945,P ＜0.01),and 10 μmol/L glycyrrhetinic acid could suppress the EGF (25 μg/L)-induced proliferation and phosphorylation of ERK1/2 in HaCaT cells.Also,both 25 μmol/L glycyrrhetinic acid and 10 μmol/L U0126 could attenuate the increase in PCNA and Notch-1 expression in HaCaT cells induced by 25 μg/L EGF.Conclusion Glycyrrhetinic acid can inhibit the EGF-induced proliferation of HaCaT cells,likely by suppressing the activation of ERK1/2 signaling pathway.
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