茶多酚对人皮肤成纤维细胞Nrf2/Bach1mRNA及核蛋白的影响

中华皮肤科杂志 ›› 2017, Vol. 50 ›› Issue (3): 199-203.

茶多酚对人皮肤成纤维细胞Nrf2/Bach1mRNA及核蛋白的影响

梁碧华¹,刘清¹,江娜¹,张倩雯¹,张尔婷¹,邓蕙妍²,李华平¹,李润祥¹,李振洁²,朱慧兰³

1. 广州市皮肤病防治所
2.
3. 广州市皮肤防治所

收稿日期:2016-03-30 修回日期:2016-12-12 发布日期:2017-03-01
通讯作者: 朱慧兰 E-mail:zhlhuilan@126.com
基金资助:
广东省2014年自然科学基金;广东省2014年科技计划项目

Effect of tea polyphenols on the mRNA and nuclear protein of Nrf2/Bach1 in human skin fibroblasts

Received:2016-03-30 Revised:2016-12-12 Published:2017-03-01
Contact: huilan zhu E-mail:zhlhuilan@126.com

摘要/Abstract

摘要： 目的探讨茶多酚对人皮肤成纤维细胞（HSF）Nrf2/Bach1 mRNA及Nrf2/Bach1核蛋白表达分布的影响。方法不同浓度茶多酚（0、2.5、5、10、20、40 mg/L）孵育HSF 24 h， MTT法检测细胞增殖活性，筛选出适宜浓度茶多酚，并用该浓度药物孵育HSF 24 h，RT?qPCR法检测Nrf2和Bach1 mRNA表达，Western印迹法检测Nrf2、Bach1核蛋白表达，免疫荧光法检测Nrf2、Bach1蛋白核分布情况。结果 MTT法检测显示，当茶多酚浓度为5 mg/L时，对HSF增殖无明显影响，故后续实验采用5 mg/L茶多酚，对照组浓度为0。HSF加入茶多酚孵育后，Bach1 mRNA相对表达量（0.629 ± 0.077）低于对照组（0.940 ± 0.033，t = 6.397，P < 0.05），Nrf2 mRNA（1.467 ± 0.076）高于对照组（0.977 ± 0.091，t = 7.133，P < 0.05）；Nrf2核蛋白表达（6.929 ± 0.121）高于对照组（3.537 ± 0.126，t = 33.636，P < 0.05），而Bach1核蛋白（1.424 ± 0.171）低于对照组（16.966 ± 1.702，t = 15.730，P < 0.05）。免疫荧光结果显示，Nrf2蛋白在细胞核内的聚集增多，Bach1蛋白在细胞核内聚集减少。结论茶多酚可促进Nrf2 mRNA、Nrf2核蛋白的表达及核分布，抑制Bach1 mRNA、Bach1核蛋白的表达和核分布，从而发挥抗氧化作用。

Abstract: Liang Bihua, Liu Qing, Jiang Na, Zhang Qianwen, Zhang Erting, Deng Huiyan, Li Huaping, Li Runxiang, Li Zhenjie, Zhu Huilan Guangzhou Institute of Dermatology, Guangzhou 510095, China Corresponding author: Zhu Huilan, Email: zhlhuilan@126.com 【Abstract】 Objective To evaluate effects of tea polyphenols on the mRNA and nucleoprotein of Nrf2/Bach1 in human skin fibroblasts （HSFs）. Methods Some HSFs were incubated with tea polyphenols at different concentrations of 0, 2.5, 5, 10, 20 and 40 mg/L for 24 hours. Methyl thiazolyl tetrazolium （MTT） assay was conducted to evaluate the proliferative activity of HSFs to screen the optimal concentration of tea polyphenols. Then, some other HSFs were treated with tea polyphenols at this optimal concentration for 24 hours. Real?time quantitative PCR （RT?qPCR） was performed to determine mRNA of Nrf2 and Bach1, Western blot analysis to measure nuclear of Nrf2 and Bach1 proteins, and immunofluorescence assay to determine the distribution of Nrf2 and Bach1 protein in the cell nucleus. Results MTT assay showed that 5 mg/L tea polyphenols had no obvious effects on the proliferation of HSFs, so 5 mg/L was chosen as the optimal concentration of tea polyphenols for subsequent experiments. HSFs cultured without tea polyphenols served as control group. After the treatment, the 5?mg/L tea polyphenol group showed significantly decreased mRNA and nuclear protein of Bach1 （mRNA: 0.629 ± 0.077 vs. 0.940 ± 0.033, t = 6.397, P < 0.05; protein: 1.424 ± 0.171 vs. 16.966 ± 1.702, t = 15.730, P < 0.05）, but significantly increased mRNA and nuclear protein of Nrf2 （mRNA: 1.467 ± 0.076 vs. 0.977 ± 0.091, t = 7.133, P < 0.05; protein: 6.929 ± 0.121 vs. 3.537 ± 0.126, t = 33.636, P < 0.05） compared with the control group. Immunofluorescence assay showed increased accumulation of Nrf2 protein, but decreased accumulation of Bach1 protein in the nucleus. Conclusion Tea polyphenols can promote the mRNA and nuclear protein as well as nuclear distribution of Nrf2 , but suppress the mRNA and nuclear protein as well as nuclear distribution of Bach1, finally exerting anti?oxidative effects.

梁碧华刘清江娜张倩雯张尔婷邓蕙妍李华平李润祥李振洁朱慧兰. 茶多酚对人皮肤成纤维细胞Nrf2/Bach1mRNA及核蛋白的影响[J]. 中华皮肤科杂志, 2017,50(3):199-203. doi:

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