Escuela de Nutrición y Dietética, Facultad de Ciencias de la Salud, Universidad de Talca, Talca 3460000, Chile.
Instituto de Química de los Recursos Naturales, Universidad de Talca, Talca 3460000, Chile.
Molecules. 2017 Dec 22;23(1):6. doi: 10.3390/molecules23010006.
Glucose autoxidation has been proposed as a key reaction associated with deleterious effects induced by hyperglycemia in the eye lens. Little is known about chromophores generated during glucose autoxidation. In this study, we analyzed the effect of oxidative and dicarbonyl stress in the generation of a major chromophore arising from glucose degradation (GDC) and its association with oxidative damage in lens proteins. Glucose (5 mM) was incubated with H₂O₂ (0.5-5 mM), Cu (5-50 μM), glyoxal (0.5-5 mM) or methylglyoxal (0.5-5 mM) at pH 7.4, 5% O₂, 37 °C, from 0 to 30 days. GDC concentration increased with incubation time, as well as when incubated in the presence of H₂O₂ and/or Cu, which were effective even at the lowest concentrations. Dicarbonylic compounds did not increase the levels of GDC during incubations. ¹H, C and FT-IR spectra from the purified fraction containing the chromophore (detected by UV/vis spectroscopy) showed oxidation products of glucose, including gluconic acid. Lens proteins solutions (10 mg/mL) incubated with glucose (30 mM) presented increased levels of carboxymethyl-lysine and hydrogen peroxide that were associated with GDC increase. Our results suggest a possible use of GDC as a marker of autoxidative reactions occurring during lens proteins glycation induced by glucose.
葡萄糖自动氧化已被提出作为与高血糖在眼睛晶状体中引起的有害影响相关的关键反应。关于葡萄糖自动氧化过程中产生的生色团知之甚少。在这项研究中,我们分析了氧化应激和二羰基应激在葡萄糖降解产生的主要生色团(GDC)的生成中的作用,以及其与晶状体蛋白氧化损伤的关系。将葡萄糖(5 mM)在 pH 7.4、5%O₂、37°C 下与 H₂O₂(0.5-5 mM)、Cu(5-50 μM)、乙二醛(0.5-5 mM)或甲基乙二醛(0.5-5 mM)孵育 0 至 30 天。孵育时间的延长以及 H₂O₂和/或 Cu 的存在会增加 GDC 的浓度,即使在最低浓度下也有效。二羰基化合物在孵育过程中不会增加 GDC 的水平。从含有生色团(通过紫外/可见光谱检测到)的纯化部分的 ¹H、C 和傅里叶变换红外(FT-IR)光谱显示葡萄糖的氧化产物,包括葡萄糖酸。用葡萄糖(30 mM)孵育的晶状体蛋白溶液(10 mg/mL)表现出增加的羧甲基赖氨酸和过氧化氢水平,这与 GDC 的增加有关。我们的结果表明,GDC 可能可用作葡萄糖诱导的晶状体蛋白糖化过程中发生的自动氧化反应的标志物。