EA 921 SONAS, Université d'Angers, SFR QUASAV 4207, Angers 49045, France.
Molecules. 2013 Nov 19;18(11):14320-39. doi: 10.3390/molecules181114320.
Advanced glycation end-products (AGEs) are involved in the pathogenesis of numerous diseases. Among them, cellular accumulation of AGEs contributes to vascular complications in diabetes. Besides using drugs to lower blood sugar, a balanced diet and the intake of herbal products potentially limiting AGE formation could be considered beneficial for patients' health. The current paper presents a simple and cheap high-throughput screening (HTS) assay based on AGE fluorescence and suitable for plant extract screening. We have already implemented an HTS assay based on vesperlysines-like fluorescing AGEs quickly (24 h) formed from BSA and ribose under physiological conditions. However, interference was noted when fluorescent compounds and/or complex mixtures were tested. To overcome these problems and apply this HTS assay to plant extracts, we developed a technique for systematic quantification of both vesperlysines (λ(exc) 370 nm; λ(em) 440 nm) and pentosidine-like (λ(exc) 335 nm; λ(em) 385 nm) AGEs. In a batch of medicinal and food plant extracts, hits were selected as soon as fluorescence decreased under a fixed threshold for at least one wavelength. Hits revealed during this study appeared to contain well-known and powerful anti-AGE substances, thus demonstrating the suitability of this assay for screening crude extracts (0.1 mg/mL). Finally, quercetin was found to be a more powerful reference compound than aminoguanidine in such assay.
糖基化终产物（AGEs）参与了许多疾病的发病机制。其中，AGEs 在细胞内的积累导致了糖尿病的血管并发症。除了使用药物降低血糖外，平衡饮食和摄入可能限制 AGE 形成的草药产品也被认为对患者的健康有益。本文提出了一种基于 AGE 荧光的简单、廉价的高通量筛选（HTS）检测方法，适用于植物提取物的筛选。我们已经实现了一种基于 BSA 和核糖在生理条件下快速（24 小时）形成的类 vesperlysine 荧光 AGE 的 HTS 检测方法。然而，当测试荧光化合物和/或复杂混合物时，会出现干扰。为了克服这些问题并将这种 HTS 检测方法应用于植物提取物，我们开发了一种系统定量测定 vesperlysine（λ(exc)370nm；λ(em)440nm）和戊糖素样（λ(exc)335nm；λ(em)385nm）AGEs 的技术。在一批药用和食用植物提取物中，只要至少一个波长的荧光降低到固定阈值以下，就会选择出现的荧光作为“命中”。在这项研究中发现的“命中”物质似乎含有众所周知的、强大的抗 AGE 物质，因此证明了该检测方法适用于粗提取物（0.1mg/mL）的筛选。最后，发现槲皮素在这种检测方法中比氨基胍更有效。
