Novel cell culture medium for use in oxidation experiments provides insights into mechanisms of endothelial cell-mediated oxidation of LDL.
Novel cell culture medium for use in oxidation experiments provides insights into mechanisms of endothelial cell-mediated oxidation of LDL.
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用于氧化实验的新型细胞培养基提供了对内皮细胞介导的 LDL 氧化机制的见解。
DOI：
10.1007/bf02577525
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Harrison,EH
中科院分区：
文献类型：
--
作者：
Dugas,TR;Morel,DW;Harrison,EH
尽管动脉粥样硬化形成的一个重要理论涉及一种血管细胞类型对血管壁内低密度脂蛋白 (LDL) 的自由基氧化，但细胞介导的 LDL 氧化的机制仍不清楚[sn1]。在这些研究中，我们检查了介质酚、硫醇和金属对内皮细胞介导的氧化的影响。我们发现细胞培养基，如 Dulbecco 改良 Eagle 培养基和基本必需培养基，无法支持细胞介导的 LDL 氧化，因为它们含有高浓度的酚红 (PR) 和酪氨酸，这两种物质都强烈抑制细胞介导的氧化。 Ham 的 F-10 是细胞介导的氧化实验常用的培养基，但也并不完全合适，因为它同时含有 PR 和半胱氨酸。半胱氨酸对于内皮细胞介导的氧化并不重要，但在细胞不存在的情况下确实会增加 LDL 的氧化。最后，对细胞介导的氧化最重要的是微摩尔浓度的 Fe(II) 或生理浓度的全铜蓝蛋白（血浆中携带铜的蛋白质）的存在。因此，用于细胞介导的氧化实验的合适培养基应含有微摩尔浓度的 Fe(II) 或生理浓度的全铜蓝蛋白，并且应不含 PR、半胱氨酸或大浓度的酪氨酸，所有这些都在此显示可抑制内皮细胞介导的 LDL 氧化。这些结果与涉及芬顿型化学和金属氧化还原循环的细胞介导的氧化机制一致。
Though one prominent theory of atherogenesis involves free-radical oxidation of low-density lipoprotein (LDL) within the vessel wall by one of the vascular cell types, the mechanism for cell-mediated LDL oxidation remains unclear[sn1]. In these studies we examined the effects of media phenols, thiols, and metals on endothelial cell-mediated oxidation. We found that cell culture media such as Dulbecco modified Eagle medium and minimal essential medium are unable to support cell-mediated oxidation of LDL because they contain high concentrations of phenol red (PR) and tyrosine, both of which strongly inhibit cell-mediated oxidation. Ham's F-10, a commonly used medium for cell-mediated oxidation experiments, is also not entirely appropriate, as it contains both PR and cysteine. Cysteine is not critical for endothelial cell-mediated oxidation, but does increase oxidation of LDL in the absence of cells. Finally, of utmost importance to cell-mediated oxidation was the presence of either micromolar concentrations of Fe(II) or physiological concentrations of holo-ceruloplasmin, the protein which carries copper in plasma. An appropriate culture medium for use in cell-mediated oxidation experiments should thus contain either micromolar concentrations of Fe(II) or physiological concentrations of holo-ceruloplasmin, and should be prepared without PR, cysteine, or large concentrations of tyrosine, all of which are shown here to inhibit endothelial cell-mediated LDL oxidation. These results are consistent with a mechanism of cell-mediated oxidation involving Fenton-type chemistry and redox cycling of the metal.