Synergistic interactions of antioxidant nutrients in a biological model system.
OBJECTIVE
The antioxidant activity of fat- and water-soluble antioxidant nutrients and their interactions in physiologic concentrations were determined in an in vitro biological model system.


METHODS
Reconstituted human serum consisting of delipidized human serum (DHS) combined with phosphatidylcholine liposomes (PCL) was used to determine antioxidant activities of physiologic concentrations of antioxidant nutrients. Radicals were initiated with 2,2'-azobis(4-methoxy-2,4-dimethylvaleronitrile) (2mmol/L), and oxidation was monitored by 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid. Fat-soluble antioxidant nutrients were incorporated into the PCL prepared by sonication and suspended in DHS to avoid any interference by the endogenous fat-soluble antioxidants. Water-soluble antioxidants were added directly into the DHS. The oxidation kinetics were monitored every 5 min up to 210 min using a microplate reader (excitation wavelength 500 nm, emission wavelength 520 nm).


RESULTS
We confirmed the synergistic protective effect of the combination of ascorbic acid (1-5 microM) and alpha-tocopherol (1-5 mircoM) against the oxidation of DHS with PCL. Furthermore, physiologic concentrations of 1) beta-carotene (0.1, 0.5 microM) and alpha-tocopherol (2.5, 5.0 microM), 2) beta-carotene (0.1, 0.5 microM) and ascorbic acid (2.5 microM), and 3) uric acid (10 uM) and alpha-tocopherol (2.5, 5.0 microM) synergistically protected oxidation of reconstituted human serum.


CONCLUSIONS
The present study results suggest a wide antioxidant network between water- and fat-soluble antioxidant nutrients in a biological system, although their actions in vivo warrant further study.
Yeum
Kyung-Jin
KJ
The Jean Mayer USDA-Human Nutrition Research Center on Aging, Tufts University, Boston, Massachusetts, USA. kyungjin.yeum@tufts.edu
Beretta
Giangiacomo
G
Krinsky
Norman I
NI
Russell
Robert M
RM
Aldini
Giancarlo
G
D000975:Antioxidants / Q000494:pharmacology*; D001205:Ascorbic Acid / Q000494:pharmacology*; D001372:Aza Compounds; D001391:Azo Compounds; D004357:Drug Synergism; D005227:Fatty Acids; D006801:Humans; D008081:Liposomes; D008954:Models, Biological; D009570:Nitriles; D010084:Oxidation-Reduction / Q000187:drug effects; D044967:Serum; D014527:Uric Acid / Q000494:pharmacology*; D024502:alpha-Tocopherol / Q000494:pharmacology*; D019207:beta Carotene / Q000494:pharmacology*
D000975:Antioxidants; D001372:Aza Compounds; D001391:Azo Compounds; C463799:BODIPY581; D005227:Fatty Acids; D008081:Liposomes; D009570:Nitriles; D019207:beta Carotene; C109522:V 70; D014527:Uric Acid; D024502:alpha-Tocopherol; D001205:Ascorbic Acid