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Transformation of Proanthocyanidin A2 to Its Isomers under Different Physiological pH Conditions and Common Cell Culture Medium

机译:在不同生理pH条件下和普通细胞培养基中原花青素A2向其异构体的转化

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Proanthocyanidins constitute an important class of polyphenols ubiquitously found in plants. They have been extensively studied for their antioxidant capacity and bioactivity in vitro and in animal models. However, their stability under different pH conditions and in cell culture medium has not been well documented, hi the present study, it was observed that proanthocyanidin A2 (PA2) was relatively more stable in acidic condition than in weak alkaline condition. PA2 was also quite unstable in basal-Dulbecco's Modified Eagle medium (b-DMEM medium) at 37 °C. The addition of PA2 to the cell culture medium accelerated its epimerization with a half-life of <15 min, and ethylenediaminetetraacetic acid (EDTA) could not stop the reaction. The results also demonstrated that the major isomers transformed in the weak alkaline condition or cell culture medium at 37 °C were identified as epicatechin-(4β->8; 2β->O->7)-ent-catechin (proanthocyanidin A4) and epicatechin-(4β->6; 2β->O->7)-entcatechin. The rates of transformation were dependent on the pH or the components of the medium. Therefore, the results obtained for PA2 in the cell culture bioassays, which were usually carried out for 24 h, might not represent the true activity of the original PA2. The stability and transformation of PA2 should be considered when the bioactivity of PA2 is evaluated in a given cell culture system.
机译:原花青素是植物中普遍存在的一类重要的多酚。他们已经在体外和动物模型中对其抗氧化能力和生物活性进行了广泛的研究。然而,它们在不同pH条件下和在细胞培养基中的稳定性尚未得到充分的证明。在本研究中,观察到原花青素A2(PA2)在酸性条件下比在弱碱性条件下相对更稳定。 PA2在37°C的基础Dulbecco改良的Eagle培养基(b-DMEM培养基)中也非常不稳定。向细胞培养基中添加PA2可以加速其差向异构化,其半衰期小于15分钟,并且乙二胺四乙酸(EDTA)不能停止反应。结果还表明,在弱碱性条件下或在37°C的细胞培养基中转化的主要异构体被鉴定为表儿茶素-(4β-> 8;2β-> O-> 7)-儿茶素(原花青素A4)和表儿茶素-(4β-> 6;2β-> O-> 7)-儿茶素。转化率取决于pH值或培养基成分。因此,通常在24小时的细胞培养生物分析中获得的PA2结果可能并不代表原始PA2的真实活性。在给定的细胞培养系统中评估PA2的生物活性时,应考虑PA2的稳定性和转化。

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