面对日益增长的抗糖化原料筛选需求，现有的体外生化抗糖化评价体系、抗糖化细胞模型和人体功效评价方法因周期长或成本高等问题，不适用于原料的大规模初筛。实验周期短、成本低、操作简便、重复性好用于抗糖化原料初步筛选的评价方法亟待确立。基于热糖化法对体外生化抗糖化评价体系进行了优化，通过检测阿魏酸、原花青素、芦丁、L-抗坏血酸等8种原料进行了抑制荧光性AGEs （Advanced glycation end products）、降解荧光性AGEs原料筛选适用性探究。并将筛选结果与已发表研究、检测AGEs降解能力的AGEs-胶原蛋白交联破坏能力实验结果进行对比，对体系筛选能力、优势和局限性进行了讨论分析。确立了优化后的反应体系为：20 mg/mL牛血清白蛋白、1 mol/L葡萄糖、0.1 mol/L PBS缓冲液组成的反应体系下，50 ℃、孵育48 h后，在λex/λem=370/452 nm波长下进行荧光强度的检测。在抑制/降解荧光性AGEs的测试中，测试2%的阿魏酸、原花青素、芦丁溶液抑制率分别达到56%，86%，95%，具有较好的抗糖化效果，阿魏酸、芦丁表现出潜在降解AGEs的趋势。优化后的体系，虽然在降解荧光性AGEs的应用上，相较于AGEs-胶原蛋白交联破坏能力测试的方法，在对照体系设置及实验结果上存在一定的局限性。但在筛选抑制荧光性AGEs的原料方面，结果稳定性、真实性佳，极大缩短了试验周期，为抗糖化功效添加剂的筛选和相应产品的开发提供了快速、稳定、节约经济成本的技术平台。
In response to the increasing demand for anti-glycation ingredient screening, existing biochemical anti-glycation evaluation systems（in vitro）, anti-glycation cell models and human efficacy evaluation methods have extended lead times and high costs. They are not suitable for large-scale primary screening of ingredients. An in vitro efficacy evaluation model with short experimental lead time, low cost, simple operation, and good reproducibility for preliminary screening of anti-glycation ingredients is urgently needed. In this study, we optimized the in vitro biochemical anti-glycation evaluation system based on the thermal glycation method and investigated the suitability of the screening of AGEs （advanced glycation end products） and degraded fluorescent AGEs by testing eight ingredients, including ferulic acid, procyanidins, rutin and L-ascorbic acid. The screening results were compared with the results of the AGEs-collagen cross-linkage disruption experiments that have been published to test the degradation ability of AGEs, and the screening ability, advantages and limitations of the system were discussed and analyzed. We confirm that the better conditions in building the model are: a reaction system consisting of 20 mg/mL BSA, 1 mol/L glucose, and 0.1 mol/L PBS buffer, a reaction temperature of 50 ℃, an incubation time of 48 h, and a detection wavelength of λex/λem=370/452 nm. In the test of inhibition/degradation of fluorescent AGEs, ferulic acid, procyanidin and rutin solutions at the tested content of 2% achieve 56%, 86% and 95% inhibition, respectively, with good anti-glycation effect, and ferulic acid and rutin show a trend of potential degradation of AGEs. The optimized system, though, has some limitations in the application of degrading fluorescent AGEs, compared with the method of AGEs-collagen cross-link disruption ability test, in terms of control system setting and experimental results. However, in the screening of ingredients for the inhibition of fluorescent AGEs, the stability and authenticity of the results are good, and the test cycle is significantly shortened, which provides a rapid, stable and economic cost-saving technical platform for the screening of anti-glycation efficacy additives and the development of related products.
