High throughput assay for evaluation of reactive carbonyl scavenging capacity

Redox Biol. 2014 Jan 30:2:590-8. doi: 10.1016/j.redox.2014.01.016. eCollection 2014.

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.

Keywords: Carbonyl scavenger; Fluorescent adduct; Reactive carbonyl species; Screening assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehydes / antagonists & inhibitors
  • Aldehydes / pharmacology*
  • Animals
  • Caco-2 Cells
  • Cell Line, Tumor
  • Cysteamine / pharmacology*
  • Cysteine / pharmacology*
  • Glycation End Products, Advanced / metabolism
  • High-Throughput Screening Assays / methods*
  • Humans
  • Mercaptoethylamines / pharmacology
  • Mice
  • Pyruvaldehyde / antagonists & inhibitors
  • Pyruvaldehyde / pharmacology*
  • Sensitivity and Specificity
  • Serum Albumin, Bovine / metabolism

Substances

  • Aldehydes
  • Glycation End Products, Advanced
  • Mercaptoethylamines
  • N-(2-mercaptoethyl)-1,3-diaminopropane
  • Serum Albumin, Bovine
  • Cysteamine
  • Pyruvaldehyde
  • 4-hydroxy-2-nonenal
  • Cysteine