A comparison between the antioxidant and peroxynitrite-scavenging functions of the vitamin E metabolites alpha- and gamma-carboxyethyl-6-hydroxychromans.
Carboxyethyl-6-hydroxychromans (CEHC) are vitamin E metabolites with proposed in vitro antioxidant function. In this study we compared the antioxidant potency of the two main CEHC metabolites found in biological fluids (i.e.. alpha-CEHC and gamma-CEHC) using two different experimental models of lipid oxidation: 1) plasma diluted 1/50 vol/vol in phosphate buffered saline (PBS) exposed to 50 micro microM Cu2+ ions, and 2) LDL (100 microg of proteins) exposed to different pro-oxidants as 2.5 microM Cu2+, 1 mM of the water soluble peroxyl radical generator 2,2'-Azobis(2-amidinopropane) hydrochloride (AAPH) and human macrophages (4 x 10(5) cells). Moreover, the two CEHC homologues were assessed for the inhibitory effect on the peroxynitrite (ONOO-)-induced nitration of tyrosine (Tyr). The results showed that in the concentration range 0.015-5 microM the CEHC metabolites and the hydrosoluble analogue Trolox exert similar concentration-dependent inhibition of the Cu2+-induced lipid oxidation of plasma. After in vitro exposure to tert-butyl hydroperoxide/Fe2+, CEHC formed chromanoxyl radicals with electron spin resonance spectra matching exactly those of their parent tocopherols. The LDL oxidation induced by AAPH or Cu2+ was significantly and similarly inhibited by 1 microM of both the CEHC homologues and Trolox. gamma-CEHC showed a slight but significantly higher inhibition of the macrophage-induced low-density lipoprotein (LDL) oxidation than alpha-CEHC. Both the CEHC homologues inhibit Tyr nitration induced by ONOO. However, gamma-CEHC produced a slightly greater inhibitory effect than (alpha-CEHC through the formation of the nitrated congener 5-nitro-gamma-CEHC. In all the systems under investigation, low nanomolar concentrations of CEHC (i.e., the concentration range in the blood of subjects with normal dietary intake of vitamin E) produced feeble antioxidant effects. In conclusion, gamma-CEHC and alpha-CEHC show similar concentration-dependent inhibition of plasma and LDL lipid oxidation. gamma-CEHC has a fairly higher potency than alpha-CEHC as ONOO- scavenger through the formation of 5-nitro-gamma-CEHC. CEHC metabolites show the same in vitro antioxidant chemistry of their parent tocopherols, but the characteristic hydrophilicity of these metabolites could result in different biopotency and roles. Further studies are needed to clarify whether CEHC could contribute to the antioxidant network in biological fluids and tissues.
Galli
Francesco
F
Department of Internal Medicine, Section of Applied and Clinical Biochemistry, University of Perugia, Italy. f.galli@unipg.it
Piroddi
Marta
M
Lannone
Anna
A
Pagliarani
Silvia
S
Tomasi
Aldo
A
Floridi
Ardesio
A
D000975:Antioxidants / Q000494:pharmacology*; D002839:Chromans / Q000378:metabolism / Q000494:pharmacology*; D003300:Copper / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy; D016166:Free Radical Scavengers / Q000494:pharmacology*; D015227:Lipid Peroxidation / Q000187:drug effects; D008077:Lipoproteins, LDL / Q000737:chemistry; D030421:Peroxynitrous Acid / Q000494:pharmacology*; D011422:Propionates / Q000378:metabolism / Q000494:pharmacology*; D014443:Tyrosine / Q000737:chemistry
D000975:Antioxidants; D002839:Chromans; D016166:Free Radical Scavengers; D008077:Lipoproteins, LDL; D011422:Propionates; D030421:Peroxynitrous Acid; C107690:2,7,8-trimethyl-2-(beta-carboxyethyl)-6-hydroxychroman; C084852:2,5,7,8-tetramethyl-2-(2'-carboxyethyl)-6-hydroxychroman; D014443:Tyrosine; D003300:Copper; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid