Evaluation of in vitro effects of natural substances of plant origin using a model of protein glycoxidation.

In an in vitro model with purified porcine aspartate aminotransferase (AST, EC 2.6.1.1) as the protein, the effects of phenolic antioxidants of plant origin (arbutin, methylarbutin, ferulic and isoferulic acids, o-coumaric and p-coumaric acids, quinic acid), flavonoids (baicalin and baicalein), and of hydroxycitric acid (HCA) at 0.5-50 mM concentration on the enzyme activities and on its glycation by 50 mM D-fructose as the glycating agent were studied. During incubation with AST at 37 degrees C up to 24 days, fructose alone decreased AST activities as a result of protein glycation. In the absence of fructose, 50 mM phenolic compounds gradually decreased AST activity, while no or a weak effect of individual compounds was found at 3 mM concentration. A direct negative effect on AST was pronounced with ferulic acid. On the other hand, beneficial influences of phenolic compounds on glycation of AST by fructose were found mostly at 3 mM concentration. Effects on glycation were vague at 50 mM concentration, probably due to a combination of direct negative influences and antiglycation effects of individual compounds. No effect, neither positive nor negative, on AST activity and protein glycation, was found with quinic acid. The flavonoid baicalin and its aglycon baicalein rapidly decreased the in vitro activity of the enzyme in all concentrations used (0.5-3 mM), and no beneficial effects of the compounds on glycation of the enzyme by fructose were found. The influence of HCA on glycation was concentration-depended, ranging from beneficial inhibition of glycation at 2.5 mM concentration to a strong decrease in AST activity at 10 mM HCA. Both the beneficial and undesirable effects of natural antioxidants should be considered in case they are used as antiglycation factors. The results obtained can contribute to the evaluation of quality of various generally recommended antioxidants.