Oxidative status of human low density lipoprotein isolated by anion-exchange high-performance liquid chromatography--assessment by total hydroxyoctadecadienoic acid, 7-hydroxycholesterol, and 8-iso-prostaglandin F(2alpha) |
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| Authors: | Kitano Soichi Yoshida Yasukazu Kawano Katsumi Hibi Nozomu Niki Etsuo |
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| Affiliation: | a Technology Development Department, SRL Inc., 153 Komiya, Hachioji, Tokyo 192-0031, Japan
b Human Stress Signal Research Center (HSSRC), National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan |
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| Abstract: | This study aims to measure the oxidative status of LDL from human plasma (n=26) as assessed by biomarkers for lipid peroxidation, total hydroxyoctadecadienoic acid (tHODE), 7alpha- and 7beta-hydroxycholesterol (t7-OHCh), and 8-iso-prostaglandin F(2alpha) (t8-iso-PGF(2alpha)) after subfractionation of LDL with an anion-exchange HPLC (AE-HPLC). LDL was separated and quantified by AE-HPLC as LDL-1, LDL-2, and LDL-3 in the order of the anionic charge of the LDL particles. The concentrations of tHODE, t7-OHCh, and t8-iso-PGF(2alpha) in both plasma and LDL subfractions were assessed after reduction and saponification. In this method, the free and ester forms of hydroperoxides, ketones, and hydroxides of linoleic acid and cholesterol are measured as tHODE and t7-OHCh, respectively. It was found that tHODE significantly correlated with the proportion of LDL-2 and LDL-3 as well as with the concentration of malondialdehyde-modified LDL in plasma. Further, by the analyses of LDL subfractions, the concentrations of tHODE, t8-iso-PGF(2alpha), and t7-OHCh in LDL-3 were found to be significantly higher than those in LDL-1 and LDL-2. These results clearly indicate that the extent of oxidation increases in the order of LDL-1
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| Keywords: | AE-HPLC, anion-exchange high-performance liquid chromatography BSTFA, N,O-bis(trimethylsilyl)trifluoroacetamide GOT, glutamic oxaloacetic transaminase GPT, glutamic pyruvic transaminase γ-GTP, γ-glutamyl transpeptidase HDL, high density lipoprotein 13-(Z,E)-HODE, 13-hydroxy-9(Z),11(E)-octadecadienoic acid 13-(E,E)-HODE, 13-hydroxy-9(E),11(E)-octadecadienoic acid 9-(E,Z)-HODE, 9-hydroxy-10(E),12(Z)-octadecadienoic 9-(E,E)-HODE, 9-hydroxy-10(E),12(E)-octadecadienoic acid 9-HODE-d4, 9S-hydroxy-10E,12Z-octadecadienoic-9,10,12,13-d4 acid HPODE, hydroperoxyoctadecadienoic acid t8-iso-PGF2α, total 8-iso-prostaglandin F2α 8-iso-PGF2α-d4, 8-iso-prostaglandin F2α-d4 LDL, low density lipoprotein Lp(a), lipoprotein (a) PBS, phosphate-buffered saline oxLDL, oxidized LDL t20:4, total arachidonate tCh, total cholesterol t18:2, total linoleate t7-OHCh, total 7-hydroxycholesterol TG, triglyceride tHODE, total hydroxyoctadecadienoic acid MDA-LDL, malondialdehyde-modified LDL PUFA, polyunsaturated fatty acid αT, α-tocopherol ZE/EE, stereoisomer ratio of HODE (9- and 13-(Z,E)-HODE/9- and 13-(E,E)-HODE) |
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