Background: while antiglycative capacity has been attributed to (poly)phenols, the exact mechanism of action remains unclear. Studies so far are often relying on supra-physiological concentrations and use of non-bioavailable compounds.
Methods: to inform the design of a physiologically relevant in vitro study, we carried out a systematic literature review of dietary interventions reporting plasma concentrations of polyphenol metabolites. Bovine Serum Albumin (BSA) was pre-treated prior to in vitro glycation: either no treatment (native), pre-oxidised (incubated with 10 nM H2O2, for 8 hours) or incubated with a mixture of phenolic acids at physiologically relevant concentrations, for 8 hours). In vitro glycation was carried out in the presence of (i) glucose only (0, 5 or 10 mM), (ii) glucose (0, 5 or 10 mM) plus H2O2 (10 nM), or (iii) glucose (0, 5 or 10 mM) plus phenolic acids (10-160 nM). Fructosamine was measured using the nitro blue tetrazolium method.
Results: following (high) dietary polyphenol intake, 3-hydroxyphenylacetic acid is the most abundant phenolic acid in peripheral blood (up to 338 μM) with concentrations of other phenolic acids ranging from 13 nM to 200 μM. The presence of six phenolic acids with BSA during in vitro glycation did not lower fructosamine formation. However, when BSA was pre-incubated with phenolic acids, significantly lower concentration of fructosamine was detected under glycoxidative conditions (glucose 5 or 10 mM plus H2O2 10 nM) (p < 0.001 vs. native BSA).
Conclusion: protein pre-treatment, either with oxidants or phenolic acids, is an important regulator of subsequent glycation in a physiologically relevant system. High quality in vitro studies under conditions closer to physiology are feasible and should be employed more frequently.