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首页> 外文期刊>Japanese Journal of Ophthalmology >Vitamins inhibit oxidant-induced apoptosis of corneal endothelial cells
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Vitamins inhibit oxidant-induced apoptosis of corneal endothelial cells

机译：维生素抑制氧化剂诱导的角膜内皮细胞凋亡

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Purpose: To determine the effects of vitamins A, C, and E supplementation on lipid peroxidation and apoptosis in corneal endothelial cells. Methods: Murine corneal endothelial cells were maintained in tissue culture medium supplemented with free iron ions, known to lead to increased lipid peroxidation. The concentration of antioxidative vitamins (ascorbic acid, tocopherol, and retinoic acid) in the cells and supernatant was determined using reversed-phase high-performance liquid chromatography. Apoptosis was assessed by quantification of caspase-3-like activity, using annexin-V/propidium iodide stains for flow cytometry. Lipid peroxidation was assessed using the malondialdehyde method. Supplementation of antioxidative vitamins was tested in the setting of apoptosis. Results: Increasing levels of free iron led to a rapid loss of antioxidative vitamins in the supernatant and corneal endothelial cells. This was correlated with rising levels of malondialdehyde and increased apoptosis. Supplementation with ascorbic acid or alpha-tocopherol alone was not sufficient to prevent lipid peroxidation in the cells, whereas a combination of vitamins C and E was able to do so. In contrast, supplementation with vitamin A alone significantly reduced oxidative stress and apoptosis. Conclusions: We present an in vitro model to test the direct influence of vitamin supplementation on corneal endothelial cells with regard to lipid peroxidation and apoptosis. We show that supplementation with antioxidative vitamins of corneal endothelial cells significantly prevents the generation of free-radical injury, lipid peroxidation, and consequent apoptosis.
机译：目的：确定补充维生素A，C和E对角膜内皮细胞脂质过氧化和凋亡的影响。方法：将小鼠角膜内皮细胞保存在补充有游离铁离子的组织培养基中，已知该游离铁离子会增加脂质过氧化作用。使用反相高效液相色谱法测定细胞和上清液中抗氧化维生素（抗坏血酸，生育酚和视黄酸）的浓度。使用膜联蛋白-V /碘化丙啶染色剂进行流式细胞术，通过定量胱天蛋白酶3样活性来评估细胞凋亡。使用丙二醛法评估脂质过氧化。在细胞凋亡的环境中测试了抗氧化维生素的补充。结果：游离铁水平的增加导致上清液和角膜内皮细胞中抗氧化维生素的快速损失。这与丙二醛水平升高和细胞凋亡增加有关。单独补充抗坏血酸或α-生育酚不足以防止细胞中的脂质过氧化，而维生素C和E的组合能够做到这一点。相反，单独补充维生素A可显着降低氧化应激和细胞凋亡。结论：我们提出了一个体外模型，以测试补充维生素对角膜内皮细胞脂质过氧化和凋亡的直接影响。我们表明，补充抗氧化维生素的角膜内皮细胞可显着防止自由基损伤，脂质过氧化和随后的细胞凋亡的产生。

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