Choice of DMEM, formulated with or without pyruvate, plays an important role in assessing the in vitro cytotoxicity of oxidants and prooxidant nutraceuticals
Choice of DMEM, formulated with or without pyruvate, plays an important role in assessing the in vitro cytotoxicity of oxidants and prooxidant nutraceuticals
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DOI：
10.1007/s11626-008-9168-z
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发表时间：
2009-05-01
影响因子：
2.1
通讯作者：
Schuck, A. G.
中科院分区：
文献类型：
--
作者：
Babich, H.;Liebling, E. J.;Schuck, A. G.
人们对营养保健品的积极健康作用非常感兴趣，特别是多酚，它具有抗氧化和促进氧化的特性。丙酮酸是一种过氧化氢的清除剂，是一些（但不是全部）商业细胞培养基配方中的一种成分，尤其是Dulbecco改良的Eagle培养基。该研究表明，与不含丙酮酸的相同培养基相比，过氧化氢、过氧化叔丁基和各种促氧化营养品对人成纤维细胞的细胞毒性在含有丙酮酸的Dulbecco改良Eagle’s培养基中有所降低。与不含丙酮酸的培养基相比，用过氧化氢处理的细胞内谷胱甘肽不受丙酮酸修饰Eagle培养基中过氧化氢处理的细胞内谷胱甘肽的影响。在这些研究中，通过测定还原性谷胱甘肽被5,5'-二硫代比斯（2-硝基苯甲酸）氧化为谷胱甘肽二硫化，在412 nm处形成黄色色原，5 -硫代-2-硝基苯甲酸，并通过荧光染料cell Tracker (TM) Green 5-氯甲基荧光素二乙酸染色的细胞中还原性谷胱甘肽的可视化，分析了酸溶性细胞提取物中的细胞内谷胱甘肽。对各种细胞培养基的调查，配制与不配制丙酮酸，证实添加过氧化氢的水平大大减少了那些培养基配制丙酮酸。本研究建议在实验设计中明确Dulbecco改良Eagle培养基的丙酮酸状态，特别是在涉及氧化应激的研究中。
There is much interest in the positive health effects of nutraceuticals, in particular, polyphenols, which have both antioxidant and prooxidant characteristics. Pyruvate, a scavenger of hydrogen peroxide, is a component in some, but not in all, commercial formulations of cell culture media, Dulbecco's modified Eagle's medium in particular. This study showed that the cytotoxicities to human fibroblasts of hydrogen peroxide, tert-butyl hydroperoxide, and various prooxidant nutraceuticals were lessened in Dulbecco's modified Eagle's medium formulated with pyruvate, as compared to the same medium but formulated without pyruvate. Intracellular glutathione was unaffected in cells treated with hydrogen peroxide in Dulbecco's modified Eagle's medium formulated with pyruvate, as compared to medium formulated without pyruvate. In these studies, intracellular glutathione was analyzed in acid-soluble cell extracts by determining the oxidation of reduced glutathione by 5,5'-dithiobis(2-nitrobenzoic acid) to glutathione disulfide, with the formation of the yellow chromagen, 5-thio-2-nitrobenzoic acid, measured spectrophotometrically at 412 nm and by the visualization of reduced glutathione in cells stained with the fluorescent dye, Cell Tracker (TM) Green 5-chloromethylfluorescein diacetate. A survey of various cell culture media, formulated with and without pyruvate, confirmed that the level of added hydrogen peroxide was greatly lessened in those media formulated with pyruvate. This study suggested that the pyruvate status of Dulbecco's modified Eagle's medium be specified in the experimental design, especially in studies involving oxidative stress.