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Yue ZHANG Hai-peng LV Cheng-ying MA Li GUO Jun-feng TAN Qun-hua PENG Zhi LIN 《Journal of Zhejiang University. Science. B》2015,16(2):103-112
目的:从茶树中克隆一种可以催化表没食子儿茶素没食子酸酯(EGCG)生成甲基化EGCG的酶——咖啡酰辅酶A氧甲基转移酶(CCo AOMT),实现甲基化EGCG的酶学合成,为甲基化EGCG的进一步开发利用提供理论依据和技术指导。创新点:本研究首次从茶树中克隆了一条CCo AOMT基因组序列;分析了CCo AOMT基因在不同茶树品种和不同成熟度茶鲜叶中的基因表达规律;证明了CCo AOMT具有催化合成甲基化EGCG的生物活性。方法:采用聚合酶链式反应(PCR)和序列分析获得CCo AOMT的编码序列和基因组序列;采用高效液相色谱-四级杆-飞行时间串联质谱技术(HPLC-QTOF-MS)分析酶促反应生成的甲基化EGCG产物(图4);采用实时荧光定量PCR分析CCo AOMT基因的表达差异(图5)。结论:本研究从茶树中克隆了CCo AOMT基因的编码序列(738 bp)和基因组序列(2678 bp),明确了该基因具有4个内含子和5个外显子;揭示了CCo AOMT可以催化EGCG生成EGCG4"Me、EGCG3"Me和EGCG3’Me等多种甲基化产物;证明了CCo AOMT具有催化生成甲基化EGCG的活性;并发现该基因的表达量高低与茶鲜叶的成熟度呈正相关关系。 相似文献
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目的：探讨EGCG对女性皮肤抗衰老效果及安全性。方法：收集88名健康志愿者作为本研究的研究对象,所有志愿者每天早晚用韩后有机茶蕊嫩白洁面乳洁面后,各使用一次、韩后有机茶蕊嫩白水、韩后有机茶蕊极嫩精华30ml、韩后有机茶蕊嫩白乳或嫩白霜,连续12周,且在此期间不使用其他护肤品,并分别于使用前、使用后6周、使用后12周测量志愿者的皮肤干燥程度、脱屑程度,皮肤角质层含水量,分析皮肤纹理及是否出现不良反应。结果：使用含有EGCG成分的护肤品前皮肤表面脱落的角质细胞数量为62134.91±7692.34,显著性高于使用6周、12周后的21098.234±6582.14、8134.974±5989.38（P〈0.05）。使用含有EGCG成分的护肤品前皮肤角质层含水量为49.234±19.29,与使用后的49.114±25.12、53.764±12.94无差异性（P〉0.05）。皮肤纹理分析：轮廓线的平均深度与平均粗糙程度在使用前后存在较大的差异性（P〈0.05）。结论：合有EGCG成分的护肤品抗衰老效果明显且没有副作用。 相似文献
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采用高效液相色谱法对不同茶叶中的表没食子儿茶素没食子酸酯(EGcG)的含量进行分析.采用C18色谱柱(4.6mm×250 mm,5μm),流动相为甲醇:水:乙酸(23:75:2,v/v),流速1.0 mL·min-1,柱温35℃,检测波长为272nm.EGCG线性范围为0.11～1.71μm,相关系数r=0.9996,平均回收率98.7%,RSD为0.7%.本方法分析速度快,灵敏度高,分离效果好,可作为测定茶叶中EGCG含量的方法. 相似文献
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Jun-guo Yang Hai-ning Yu Shi-li Sun Lan-cui Zhang Guo-qing He Undurti N. Das Hui Ruan Sheng-rong Shen 《Journal of Zhejiang University. Science. B》2009,10(6):411-421
Objective: To evaluate effects of epigallocatechin-3-gallate (EGCG) on the viability, membrane properties, and zinc distribution, with and without the presence of Zn2+, in human prostate carcinoma LNCaP cells. Methods: We examined changes in cellular morphology and membrane fluidity of LNCaP cells, distribution of cellular zinc, and the incorporated portion of EGCG after treatments with EGCG, Zn2+, and EGCG+Zn2+. Results: We observed an alteration in cellular morphology and a decrease in membrane fluidity of LNCaP cells after treatment with EGCG or Zn2+. The proportion of EGCG incorporated into liposomes treated with the mixture of EGCG and Zn2+ at the ratio of 1:l was 90.57%, which was significantly higher than that treated with EGCG alone (30.33%). Electron spin resonance (ESR) studies and determination of fatty acids showed that the effects of EGCG on the membrane fluidity of LNCaP were decreased by Zn2+. EGCG accelerated the accumulation of zinc in the mitochondria and cytosol as observed by atomic absorption spectrometer. Conclusion: These results show that EGCG interacted with cell membrane,decreased the membrane fluidity of LNCaP cells, and accelerated zinc accumulation in the mitochondria and cytosol, which could be the mechanism by which EGCG inhibits proliferation of LNCaP cells. In addition, high concentrations of Zn2+ could attenuate the actions elicited by EGCG. 相似文献
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研究了没食子儿茶素没食子酸酯（EGCG）对人肝癌细胞BEL-7402增殖凋亡及对信号转导蛋白和转录激活因子3（Stat3）蛋白及磷酸化Stat3蛋白表达的影响.应用MTT法检测不同浓度EGCG对BEL-7402细胞增殖的抑制作用,用流式细胞仪分析细胞的凋亡率,用ELISA方法检测EGCG对Stat3蛋白及磷酸化的Stat3蛋白表达的影响.结果表明：EGCG有抑制肝癌细胞增殖促进凋亡的作用,并呈剂量依赖性.ELISA结果显示EGCG能降低磷酸化Stat3蛋白的表达水平.EGCG能抑制肝癌细胞的增殖促进凋亡,其机制可能有与下调磷酸化的Stat3蛋白的表达有关. 相似文献
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Tea polyphenols have been shown to have anticancer activity in many studies.In the present study,we investigated effects of theaflavin-3-3'-digallate(TF3),one of the major theaflavin monomers in black tea,in combination with ascorbic acid(AA),a reducing agent,and(-)-epigallocatechin-3-gallate(EGCG),the main polyphenol presented in green tea,in combination with AA on cellular viability and cell cycles of the human lung adenocarcinoma SPC-A-1 cells.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) assay showed that the 50% inhibition concentrations(IC50) of TF3,EGCG,and AA on SPC-A-1 cells were 4.78,4.90,and 30.62 μmol/L,respectively.The inhibitory rates of TF3 combined with AA(TF3+AA) and EGCG combined with AA(EGCG+AA) at a molar ratio of 1:6 on SPC-A-1 cells were 54.4% and 45.5%,respectively.Flow cytometry analysis showed that TF3+AA and EGCG+AA obviously increased the cell population in the G0/G1 phase of the SPC-A-1 cell cycle from 53.9% to 62.8% and 60.0%,respectively.TF3-treated cells exhibited 65.3% of the G0/G1 phase at the concentration of its IC50.Therefore,TF3+AA and EGCG+AA had synergistic inhibition effects on the proliferation of SPC-A-1 cells,and significantly held SPC-A-1 cells in G0/G1 phase.The results suggest that the combination of TF3 with AA or EGCG with AA enhances their anticancer activity. 相似文献
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