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Biomedicines 2018, 6(3), 88; https://doi.org/10.3390/biomedicines6030088

Improved Methods for the Rapid Formation and Prevention of Advanced Glycation End Products (AGEs) In Vitro by Coupling to the Hypoxanthine/Xanthine Oxidase Assay System

1
Instituto Federal de Educação, Ciência e Tecnologia do Ceará, Departamento de Química, Av. José de Freitas Queiroz, 5000, Quixadá–CE CEP 63902-580, Brazil
2
Campus do Pici–Bloco 935 superior–Laboratório de Produtos Naturais e 10 Biotecnologia e (LPNBio), Universidade Federal do Ceará, Fortaleza CEP 60451-970, Brazil
3
Instituto Federal de Educação, Ciência e Tecnologia do Ceará, Departamento de Química, rua Luis Cunha, S/N–Monte Castelo, Ubajara–CE CEP 62350-000, Brazil
4
National Center for Tumor Diseases (NCT)/German Cancer Research Center (DKFZ), Division of Preventive Oncology, Im Neuenheimer Feld 460, 69120 Heidelberg, Germany
*
Author to whom correspondence should be addressed.
Received: 22 July 2018 / Revised: 8 August 2018 / Accepted: 10 August 2018 / Published: 15 August 2018
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Abstract

Advanced glycation end products (AGEs) represent a set of molecules that contribute directly to the initiation and aggravation of diseases associated with ageing. AGEs are produced by the reaction between reducing sugars (or α-dicarbonyl compounds), proteins, and amino acid residues. Previous in vitro methods using non-enzymatic procedures described in the literature require an incubation period of 1–3 weeks to generate AGEs. In this study, the reaction time for the formation of AGEs (48 and 3 h) was significantly reduced by adaptation of methods previously described in the literature and coupling them to the free radical generation system termed hypoxanthine/xanthine oxidase assay. The incorporation of this assay into the experimental system accelerated the production of AGEs as a result of the formation of reactive oxygen species (ROS), as shown by increased fluorescence. The capacity of different classes of chemical compounds (aminoguanidine, chlorogenic acid, rutin, and methanol extracts of Hancornia speciosa Gomes) to inhibit protein glycation by acting as scavenging agents of α-dicarbonyl species was evaluated. Aminoguanidine and, especially, rutin identified in the leaf extracts of H. speciosa Gomes showed a high capacity to act as scavengers of reactive carbonyl species RCS-trapping, resulting in the inhibition of AGEs formation. View Full-Text
Keywords: advanced glycation end products; glyoxal; hypoxanthine/xanthine oxidase; methyl glyoxal; RCS-trapping; rutin; xanthine oxidase advanced glycation end products; glyoxal; hypoxanthine/xanthine oxidase; methyl glyoxal; RCS-trapping; rutin; xanthine oxidase
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Marques, S.; Trevisan, T.; Maia, C.; Breuer, A.; Owen, R.W. Improved Methods for the Rapid Formation and Prevention of Advanced Glycation End Products (AGEs) In Vitro by Coupling to the Hypoxanthine/Xanthine Oxidase Assay System. Biomedicines 2018, 6, 88.

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