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Tuning a 96-well microtiter plate fluorescence-based assay to identify AGE inhibitors in crude plant extracts

Séro, Luc ; Sanguinet, Lionel ; Blanchard, Patricia ; Dang, Bach Tai ; Morel, Sylvie ; Richomme, Pascal ; Séraphin, Denis ; Derbré, Séverine

Molecules (Basel, Switzerland), 2013-11, Vol.18 (11), p.14320-14339 [Periódico revisado por pares]

Switzerland: MDPI AG

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Título:
Tuning a 96-well microtiter plate fluorescence-based assay to identify AGE inhibitors in crude plant extracts
Autor: Séro, Luc ; Sanguinet, Lionel ; Blanchard, Patricia ; Dang, Bach Tai ; Morel, Sylvie ; Richomme, Pascal ; Séraphin, Denis ; Derbré, Séverine
Assuntos: advanced glycation end-products ; Arginine ; Arginine - analogs & derivatives ; Arginine - analysis ; automation ; Biochemistry, Molecular Biology ; Fluorescence ; Glycation End Products, Advanced ; Glycation End Products, Advanced - analysis ; Life Sciences ; Lysine ; Lysine - analogs & derivatives ; Lysine - analysis ; natural products ; pentosidine ; plant extract screening ; Plant Extracts ; Plant Extracts - chemistry ; vesperlysines
É parte de: Molecules (Basel, Switzerland), 2013-11, Vol.18 (11), p.14320-14339
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
PMCID: PMC6270619
Descrição: 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.
Editor: Switzerland: MDPI AG
Idioma: Inglês;Francês

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Tuning a 96-well microtiter plate fluorescence-based assay to identify AGE inhibitors in crude plant extracts

Séro, Luc ; Sanguinet, Lionel ; Blanchard, Patricia ; Dang, Bach Tai ; Morel, Sylvie ; Richomme, Pascal ; Séraphin, Denis ; Derbré, Séverine

Switzerland: MDPI AG

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