Attenuation of lipid peroxidation by antioxidants in rat-1 fibroblasts: comparison of the lipid peroxidation reporter molecules cis-parinaric acid and C11-BODIPY(581/591) 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-parinaric 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-BODIPY(581/591). To further explore this probe, we evaluated the protective effect of 12 natural antioxidants in rat-1 fibroblasts subjected to 50 microM cumene-hydroperoxide using both probes. The same pecking order for the individual antioxidant efficacies was obtained: alpha-tocopherol approximately gamma-tocopherol > quercetin approximately lycopene > kaempferol > palm oil > hydroxy-tyrosol > > alpha-carotene = beta-carotene = lutein = tyrosol = chlorogenic acid. This validates the accuracy of the C11-BODIPY(581/591) 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.
Drummen
Gregor P C
GP
Department of Biochemistry of Lipids, Institute and Graduate School of Biomembranes, Utrecht University, Padualaan 8, 3584CH Utrecht, The Netherlands.
Makkinje
Miriam
M
Verkleij
Arie J
AJ
Op den Kamp
Jos A F
JA
Post
Jan A
JA
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001896:Boron Compounds / Q000737:chemistry*; D002460:Cell Line; D005231:Fatty Acids, Unsaturated / Q000737:chemistry*; D005347:Fibroblasts / Q000187:drug effects*; D015227:Lipid Peroxidation / Q000187:drug effects*; D015335:Molecular Probes; D051381:Rats
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D000975:Antioxidants; D001896:Boron Compounds; D005231:Fatty Acids, Unsaturated; D015335:Molecular Probes; C013641:parinaric acid