Attenuation of lipid peroxidation by antioxidants in rat-1 fibroblasts:: comparison of the lipid peroxidation reporter molecules cis-parinaric acid and C11-BODIPY581/591 in a biological setting

Lipid peroxidation is a major factor in the pathogenesis of many disease states. To detect the initial stages of lipid peroxidation or evaluate antioxidant efficacy, cis-parmaric acid (cis-PnA) has been successfully used and thoroughly validated. However, cis-PnA is not very well suited for medium throughput screening of antioxidants in living cells. We recently introduced and validated a lipid peroxidation reporter molecule, C11-BODIPY581/591. To further explore this probe, we evaluated the protective effect of 12 natural antioxidants in rat-1 fibroblasts subjected to 50 muM cumene-hydroperoxide using both probes. The same pecking order for the individual antioxidant efficacies was obtained: alpha-tocopherol approximate to gamma-tocopherol>quercetin lycopene>kaempferol>palm oil>hydroxy-tyrosol>>alpha-carotene=beta-carotene=lutein=tyrosol=chlorogenic acid. This validates the accuracy of the C11-BODIPY581/591 method and shows that this assay is an accurate and highly flexible method for indexing lipid peroxidation or determining antioxidant efficacy in living cells in a medium throughput scenario. The antioxidant efficacy was compared with their one-electron reduction potential, hydrophobicity and Trolox C equivalent antioxidant capacity. Our results show that although these parameters are valuable for determining structure-function relationships, they have limited predictive value for antioxidant efficacy in vivo. (C) 2004 Elsevier B.V All rights reserved.