Figures & data
Fig. 1. Structure of OFA.
Fig. 2. DPPH radical scavenging capacity of OFA.
Notes: DPPH was mixed with OFA and Trolox in methanol and incubated for 30 min at room temperature in the dark. The absorbance of the solutions was measured at 517 nm. Data are expressed as means ± SD (n = 3). OFA (■), Trolox (●).
Fig. 3. Effect of OFA on peroxyl radical-mediated lipid peroxidation of LDL.
Notes: LDL (200 μg protein/ml) oxidation was initiated by 5 mM AAPH at 37 °C. CE-OOH was determined by HPLC with UV detection. Data are expressed as means ± SD (n = 3). Control (●), 0.5 μM OFA (■), 1 μM OFA (▲), 2 μM OFA (◆).
Fig. 4. Effect of OFA on transition metal ion-catalyzed lipid peroxidation of LDL.
Notes: LDL in PBS was oxidized by 5 μM Cu2+ at 37 °C. TBARS was analyzed by HPLC with fluorescence detection. Data are expressed as means ± SD (n = 3). Control (●), 0.5 μM OFA (■), 1 μM OFA (▲), and 2 μM OFA (◆).
Fig. 5. Effect of OFA on apo B-100 modification in oxLDL.
Notes: LDL in PBS was oxidized by Cu2+ at 37 °C for 3 h. (A) OxLDL was analyzed by SDS-PAGE and CBB staining. (B) OxLDL was determined by Western blotting using anti-human apo B-100 antibody. Immunoreactive bands were detected with ECL reagent. Lane 1, native LDL; lane 2, LDL oxidized by Cu2+; lane 3, LDL oxidized by Cu2+ in the presence of 2 μM OFA.
Fig. 6. Effect of OFA on protein carbonyl formation in oxLDL.
Notes: LDL in PBS was oxidized by Cu2+ at 37 °C for 3 h. OxLDL was treated with 2,4-dinitrophenylhydrazine and analyzed by SDS-PAGE and Western blotting using rabbit anti-dinitrophenyl antisera as primary antibody and by ECL detection. Lane 1, native LDL; lane 2, LDL oxidized by Cu2+; lanes 3, 4, and 5, LDL oxidized by Cu2+ in the presence of 0.5, 1, and 2 μM of OFA, respectively.
Fig. 7. Effect of OFA on heparin-binding activity of apo B-100 in oxLDL.
Notes: LDL in PBS was oxidized by Cu2+ at 37 °C for 3 h. OxLDL was incubated with 100 μL heparin Sepharose at 4 °C for 3 h. After washing heparin Sepharose beads with Tween-TBS, proteins bound to the beads were subjected to SDS-PAGE and CBB staining. Lane 1, native LDL; lane 2, LDL oxidized by Cu2+; lane 3, LDL oxidized by Cu2+ in the presence of 2 μM OFA.
Fig. 8. Effect of OFA on peroxynitrite-mediated lipid peroxidation of LDL.
Notes: LDL in PBS was oxidized by 1 mM SIN-1 at 37 °C. CE-OOH was analyzed by HPLC with UV detection. Data are expressed as means ± SD (n = 3). Control (●), 2 μM OFA (■), 4 μM OFA (▲), and 8 μM OFA (◆).
Fig. 9. Effect of OFA on nitrotyrosine formation in oxLDL.
Notes: LDL in PBS was oxidized by 1 mM SIN-1 at 37 °C for 4 h. (A) OxLDL was analyzed by SDS-PAGE and CBB staining. (B) OxLDL was determined by Western blotting using anti-nitrotyrosine antibody and by ECL detection. Lane 1, native LDL; lane 2, LDL oxidized by SIN-1; lane 3, LDL oxidized by SIN-1 in the presence of 8 μM OFA.
