title: High throughput assay for evaluation of reactive carbonyl scavenging capacity.
abstract: Many carbonyl species from either lipid peroxidation or glycoxidation are extremely reactive and can disrupt the function of proteins and enzymes. 4-hydroxynonenal and methylglyoxal are the most abundant and toxic lipid-derived reactive carbonyl species. The presence of these toxics leads to carbonyl stress and cause a significant amount of macromolecular damages in several diseases. Much evidence indicates trapping of reactive carbonyl intermediates may be a useful strategy for inhibiting or decreasing carbonyl stress-associated pathologies. There is no rapid and convenient analytical method available for the assessment of direct carbonyl scavenging capacity, and a very limited number of carbonyl scavengers have been identified to date, their therapeutic potential being highlighted only recently. In this context, we have developed a new and rapid sensitive fluorimetric method for the assessment of reactive carbonyl scavengers without involvement glycoxidation systems. Efficacy of various thiol- and non-thiol-carbonyl scavenger pharmacophores was tested both using this screening assay adapted to 96-well microplates and in cultured cells. The scavenging effects on the formation of Advanced Glycation End-product of Bovine Serum Albumin formed with methylglyoxal, 4-hydroxynonenal and glucose-glycated as molecular models were also examined. Low molecular mass thiols with an α-amino-β-mercaptoethane structure showed the highest degree of inhibitory activity toward both α,β-unsaturated aldehydes and dicarbonyls. Cysteine and cysteamine have the best scavenging ability toward methylglyoxal. WR-1065 which is currently approved for clinical use as a protective agent against radiation and renal toxicity was identified as the best inhibitor of 4-hydroxynonenal.
authors.affiliation: Aix Marseille Université, CNRS, ICR UMR 7273, 13397, Marseille, France.
authors.forename: N
authors.initials: N
authors.lastname: Vidal
authors.affiliation: Aix Marseille Université, CNRS, ICR UMR 7273, 13397, Marseille, France.
authors.forename: J P
authors.initials: JP
authors.lastname: Cavaille
authors.affiliation: Aix Marseille Université, CNRS, ISM2 UMR 7313, 13397, Marseille, France.
authors.forename: F
authors.initials: F
authors.lastname: Graziani
authors.affiliation: Aix Marseille Université, CNRS, ICR UMR 7273, 13397, Marseille, France.
authors.forename: M
authors.initials: M
authors.lastname: Robin
authors.affiliation: Aix Marseille Université, CNRS, ICR UMR 7273, 13397, Marseille, France.
authors.forename: O
authors.initials: O
authors.lastname: Ouari
authors.affiliation: Aix Marseille Université, CNRS, ICR UMR 7273, 13397, Marseille, France.
authors.forename: S
authors.initials: S
authors.lastname: Pietri
authors.affiliation: Aix Marseille Université, CNRS, ICR UMR 7273, 13397, Marseille, France.
authors.forename: P
authors.initials: P
authors.lastname: Stocker
mesh_terms: D000447:Aldehydes / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D000818:Animals; D018938:Caco-2 Cells; D045744:Cell Line, Tumor; D003543:Cysteamine / Q000494:pharmacology*; D003545:Cysteine / Q000494:pharmacology*; D017127:Glycation End Products, Advanced / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008624:Mercaptoethylamines / Q000494:pharmacology; D051379:Mice; D011765:Pyruvaldehyde / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000378:metabolism
chemical_list: D000447:Aldehydes; D017127:Glycation End Products, Advanced; D008624:Mercaptoethylamines; C020174:N-(2-mercaptoethyl)-1,3-diaminopropane; D012710:Serum Albumin, Bovine; D003543:Cysteamine; D011765:Pyruvaldehyde; C027576:4-hydroxy-2-nonenal; D003545:Cysteine
keywords: Carbonyl scavenger; Fluorescent adduct; Reactive carbonyl species; Screening assay