Department of Biochemistry, Faculty of Biology, University of Warmia and Mazury, Oczapowskiego 1A, 10-719, Olsztyn, Poland,
Eur J Nutr. 2013 Dec;52(8):1927-38. doi: 10.1007/s00394-013-0494-x. Epub 2013 Jan 20.
The purpose of this study was to determine the influence of Maillard reaction (MR, glycation) on biochemical and biological properties of the major peanut allergen Ara h 1.
Three different time/temperature conditions of treatment were applied (37, 60, and 145 °C). The extent of MR was assessed by SDS-PAGE, loss of free amino groups, fluorescence intensity, content of bound sugar and fructosamine. The Caco-2 model system was applied to study effects of hydrolysed and non-hydrolysed Ara h 1 on proliferation and interleukin-8 (IL-8) secretion from Caco-2 cells.
We demonstrated significant differences in the biochemical properties of Ara h 1 glycated at different time/temperature conditions. Glycation of Ara h 1 at 37 °C was shown to cause least biochemical changes, not limiting pepsin hydrolysis. Loss of free amino groups, increase of fluorescence and brown colour of Ara h 1 glycated at 60 and 145 °C indicated advanced and final stages of MR. Non-treated Ara h 1 inhibited Caco-2 cell proliferation and stimulated IL-8 secretion. This effect was less pronounced for glycated Ara h 1. Incubation of Caco-2 cells with non-hydrolysed Ara h 1, glycated at the temperature of 37 and 60 °C, did not stimulate IL-8 secretion.
Each applied time/temperature-treatment combination caused different biochemical changes of Ara h 1, underlining diversity of formed MRPs. MR, independently of temperature/time conditions, reduced the pro-inflammatory properties of native Ara h 1, reflected in stimulation of IL-8 secretion from intestinal epithelial cells.
本研究旨在确定美拉德反应（MR，糖化）对主要花生过敏原 Ara h 1 的生化和生物学特性的影响。
应用三种不同的时间/温度处理条件（37、60 和 145°C）。通过 SDS-PAGE、游离氨基基团损失、荧光强度、结合糖和果糖胺含量来评估 MR 的程度。应用 Caco-2 模型系统研究水解和未水解的 Ara h 1 对 Caco-2 细胞增殖和白细胞介素-8（IL-8）分泌的影响。
我们证明了在不同时间/温度条件下糖化 Ara h 1 的生化性质存在显著差异。在 37°C 糖化 Ara h 1 导致的生化变化最小，不会限制胃蛋白酶水解。在 60 和 145°C 糖化的 Ara h 1 的游离氨基基团损失、荧光强度增加和棕色表明 MR 处于晚期和终期阶段。未经处理的 Ara h 1 抑制 Caco-2 细胞增殖并刺激 IL-8 分泌。而未经处理的 Ara h 1 对 Caco-2 细胞增殖的抑制作用和对 IL-8 分泌的刺激作用则不那么明显。未水解的 Ara h 1 在 37 和 60°C 下糖化，孵育 Caco-2 细胞不会刺激 IL-8 分泌。
每种应用的时间/温度处理组合都会导致 Ara h 1 发生不同的生化变化，突出了形成的 MRPs 的多样性。MR 独立于温度/时间条件，降低了天然 Ara h 1 的促炎特性，反映在刺激肠上皮细胞分泌 IL-8 上。
