Fig 1.
SO4= uptake in human erythrocytes under H2O2 treatment.
Time course of SO4= uptake in human erythrocytes measured in control conditions (untreated erythrocytes) or treated with 10 μM DIDS, applied at the beginning of incubation in SO4= medium. Points represent the mean ± SEM from separate experiments (see Table 1), where ***p<0.001 significant versus control and §§p<0.01 significant versus 100 μM H2O2, as determined by one way ANOVA followed by Bonferroni's post hoc test, by comparing all values of theoretical curves, at all time points.
Fig 2.
Erythrocytes morphology under H2O2 treatment.
Light microscope observations (400x magnification) of: A) Untreated erythrocytes; B) 300 μM H2O2 treated erythrocytes at 5 min of SO4= medium incubation; C) 300 μM H2O2 treated erythrocytes at 90 min of SO4= medium incubation. Arrows indicate significant morphological changes in both B) and C) compared to the control (untreated) erythrocytes A).
Table 1.
Rate constant for SO4= uptake in treated and untreated erythrocytes.
Fig 3.
SO4= uptake in human erythrocytes under H2O2 plus GSH treatment.
Time course of SO4= uptake in human erythrocytes measured in control conditions (untreated erythrocytes) or treated with 300 μM H2O2, or treated with 2 mM GSH and then with 300 μM H2O2. Points represent the mean ± SEM from separated experiments (see Table 1), where ***p<0.001 significant versus control and ¥¥p<0.01 significant versus 300 μM H2O2, as determined by one way ANOVA followed by Bonferroni's post hoc test, by comparing all values of theoretical curves, at all time points.
Fig 4.
Erythrocytes morphology under H2O2 plus GSH treatment Light microscope observations (400x magnification) of: A) 300 μM H2O2 treated erythrocytes, observed after 5 min incubation in SO4= medium and B) erythrocytes treated with 2 mM GSH and then with 300 μM H2O2, observed after 5 min incubation in SO4= medium; C) 300 μM H2O2 treated erythrocytes, observed after 90 min incubation in SO4= medium and D) erythrocytes treated with 2 mM GSH and then with 300 μM H2O2 observed after 90 min incubation in SO4= medium. Arrows indicate that morphological changes after H2O2 treatment are not inhibited by GSH.
Fig 5.
SO4= uptake in human erythrocytes under H2O2 plus curcumin treatment.
Time course of SO4= uptake in human erythrocytes measured in control conditions (untreated erythrocytes) or treated with 300 μM H2O2 preceded or not by 10 μM curcumin application. Points represent the mean ± SEM from separate experiments (see Table 1), where ***p<0.001 significant versus control or **p<0.01 significant versus control and #p<0.05 significant versus 300 μM H2O2, as determined by one way ANOVA followed by Bonferroni's post hoc test, by comparing all values of theoretical curves, at all time points.
Fig 6.
Erythrocytes morphology under H2O2 plus curcumin treatment.
Light microscope observations (400x magnification) of: A) 300 μM H2O2 treated erythrocytes, observed after 5 min incubation in SO4= medium and B) erythrocytes treated with 10 μM curcumin followed by 300 μM H2O2, observed after 5 min incubation in SO4= medium; C) 300 μM H2O2 treated erythrocytes, observed after 90 min incubation in SO4= medium and D) erythrocytes treated with 10 μM curcumin followed by 300 μM H2O2, observed after 90 min incubation in SO4= medium. Arrows indicate morphological changes after H2O2 treatment, attenuated by curcumin treatment.
Fig 7.
MDA levels and –SH groups estimation under H2O2 treatment.
A) MDA levels observed in control (untreated erythrocytes) and in erythrocytes treated with 300 μM H2O2. Data are presented as means ± SEM from at least 3 experiments, where n.s. is not significantly different versus control, as determined by t-Student test. B) Percentage of –SH groups measured in control (untreated erythrocytes), in erythrocytes treated with either 300 μM H2O2 or 2 mM NEM. Bars represent the mean ± SEM from at least 3 experiments, where n.s. is not significant versus control, §§§ p<0.001 significant versus control and 300 μM H2O2-treated erythrocytes, as determined by one way ANOVA followed by Bonferroni's post hoc test.
Fig 8.
SO4= uptake in human hemoglobin-free resealed ghosts of erythrocytes under H2O2 treatment.
Time course of SO4= uptake in hemoglobin-free resealed ghosts of erythrocytes measured in control conditions (untreated) or treated with either 300 μμM H2O2, or 10 μ M DIDS applied at the beginning of the incubation in SO4= medium. Points represent the mean ± SEM from separate experiments (see Table 2), where ***p<0.001 significant versus control or §§§p<0.001 significant versus 10 μ M DIDS and n.s. not significant versus control, as determined by one way ANOVA followed by Bonferroni's post hoc test, by comparing all values of theoretical curves, at all time points.
Table 2.
Rate constant for SO4= uptake in treated and untreated resealed ghosts of erythrocytes.