摘要
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MTr法检测不同浓度EGF(0、1、5、10、25、50、100μg,IJ)以及不同浓度甘草次酸(0、0.1、I、10、25、50、100la,mol/L)对HaCaT细胞增殖的影响,25p,mol/L甘草次酸、10μmol/LMEKI/2抑制剂(U0126)对25μg/LEGF促HaCaT细胞增殖的抑制作用。蛋白免疫印迹法检测25μg,LEGF对ERKI/2磷酸化的促进作用,25/zmol/L甘草次酸、10/zmol/LU0126对ERKl/2磷酸化的抑制作用,25gg/LEGF、25μmol/L甘草次酸、10μmol/LU0126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SPSSl7.0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0~100μg/L范围内促进HaCaT细胞增殖(r=O.798,P〈0.05),在0~50μg/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—100μmol/L范围内浓度依赖性地抑制HaCaT细胞增殖(r=-0.945,P〈0.01);25μmol/L甘草次酸明显抑制25μg,LEGF对HaCaT细胞的促增殖作用以及对ERKl/2的促磷酸化作用。同时,25μmol/L甘草次酸、10txmol/LU0126均抑制25Ixg/LEGF对HaCaT细胞PCNA、Notch—I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ERKl/2信号通路的激活有关。
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 (MTF) 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 glyeyrrhetinic acid (0,0.1,1.0,10,25,50,100 μmol/L) alone, or the combination of 25 p.g/L EGF with 25 μmol/L glycyrrhetinic acid or 10 μmol/L U0126 (an inhibitor of MEK1/2). Western blot was carried out to measure the protein expression of proliferating cell nuclear antigen (PCNA),Notch-l, ERK 1/2 and phosphorylated ERK 1/2 in HaCaT cells treated with 25. μg/L EGF, 10 μmol/L U0126, 25 μmol/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 O-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 Ixg/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 txmol/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.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2013年第4期278-281,共4页
Chinese Journal of Dermatology
基金
浙江省自然科学基金(Y2100759)