Atherogenesis May Involve the Prooxidant and Proinflammatory Effects of Ferryl Hemoglobin
Figure 5
GSH/GPx and Hp inhibit H2O2 and oxLDL-mediated oxidation and crosslinking of Hb. ((a) and (b)) Human Hb (20 μmol/L heme) was pretreated with Hp (50 μmol/L) for 10 minutes at 37°C followed by 90 minute incubation with H2O2 (200 μmol/L) and oxLDL (400 μg/mL). Separately, H2O2 (200 μmol/L) and oxLDL (400 μg/mL) were pretreated with GSH/GPx for 10 minutes at 37°C followed by a 90-minutes of incubation with Hb (20 μmol/L heme). (a) MetHb was determined spectrophotometrically. (b) Oxidation-induced dimer formation was monitored by Western blot. Immunoblot is a representative of three independent experiments. Results are shown as mean ± S.D. () from one representative experiment of three.
