Study on the Stability of (-)-Epigallocatechin-3-gallate
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摘要: 目的：解析表没食子儿茶素没食子酸酯（（-）-epigallocatechin-3-gallate，EGCG）氧化聚合的环境条件，及其稳定性变化对体外培养细胞系氧化还原平衡的影响，为深入探讨EGCG在细胞培养体系及活体中的作用机制奠定基础。方法：在无细胞体系中，设置不同的环境条件，溶液环境（H2O、RPMI1640、DMEM）、温度（37和74℃）、EGCG浓度（0、20、40、80、320 μmol/L）、反应时间（3、6、12、24 h）、pH （5和9）等，通过检测OD578值变化评估EGCG在不同条件下氧化聚合的差异；分析EGCG氧化聚合变化对受试细胞内外H2O2水平的影响。结果：发现RPMI1640和DMEM培养液较H2O更利于EGCG的氧化聚合，高温高EGCG浓度及碱性条件促进EGCG氧化聚合，同时EGCG的氧化聚合随反应时间延长而加剧。在含有细胞的培养体系中EGCG也会发生自动氧化聚合并产生H2O2，导致细胞内外的H2O2浓度升高，但胞内H2O2浓度呈现先上升后下降的趋势。结论：溶液环境和pH变化是引起EGCG氧化聚合的关键因素，在细胞培养中20 μmol/L的EGCG可通过氧化聚合效应发挥更好的抗氧化活性。Abstract: Objective: To analyze the environmental conditions of(-)-epigallocatechin-3-gallate(EGCG)oxidative polymerization and the effects of changes in its stability on the redox balance of the test cells,and to lay the foundation for an in-depth study of the role of EGCG in cell culture systems and living organisms. Methods: The research set different conditions of solution environment,EGCG concentration,temperature,time,pH,to evaluate the difference of EGCG oxidative polymerization under different conditions by detecting the change of OD578 value. Result: The results showed that RPMI1640 and DMEM medium were more favorable for EGCG oxidation polymerization than H2O. High temperature and high EGCG concentration and alkaline conditions promoted EGCG oxidation polymerization. At the same time,the oxidative polymerization of EGCG intensified with the extension of the reaction time. In a cell-containing culture system,EGCG also underwent auto-oxidative polymerization and produced H2O2,which increased the H2O2 concentration inside and outside the cell. However,the intracellular H2O2 concentration showed a trend of first increase and then decrease. Conclusion: Solution environment and pH changes are the key factors that cause EGCG oxidative polymerization. 20 μmol/L EGCG is more conducive to exert antioxidant activity through oxidative polymerization effect in cell culture.
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Keywords:
- (-)-epigallocatechin-3-gallate /
- stability /
- oxidation /
- anti-oxidation
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