Oxidation Stress as a Mechanism of Aging in Human Erythrocytes: Protective Effect of Quercetin - PubMed (original) (raw)

Oxidation Stress as a Mechanism of Aging in Human Erythrocytes: Protective Effect of Quercetin

Alessia Remigante et al. Int J Mol Sci. 2022.

Abstract

Aging is a multi-factorial process developing through a complex net of interactions between biological and cellular mechanisms and it involves oxidative stress (OS) as well as protein glycation. The aim of the present work was to verify the protective role of Quercetin (Q), a polyphenolic flavonoid compound, in a d-Galactose (d-Gal)-induced model of aging in human erythrocytes. The anion-exchange capability through the Band 3 protein (B3p) measured by the rate constant of the SO42- uptake, thiobarbituric acid reactive substances (TBARS) levels-a marker of lipid peroxidation-total sulfhydryl (-SH) groups, glycated hemoglobin (A1c), and a reduced glutathione/oxidized glutathione (GSH-GSSG) ratio were determined following the exposure of erythrocytes to 100 mM d-Gal for 24 h, with or without pre-incubation with 10 µM Q. The results confirmed that d-Gal activated OS pathways in human erythrocytes, affecting both membrane lipids and proteins, as denoted by increased TBARS levels and decreased total sulfhydryl groups, respectively. In addition, d-Gal led to an acceleration of the rate constant of the SO42- uptake through the B3p. Both the alteration of the B3p function and oxidative damage have been improved by pre-treatment with Q, which preferentially ameliorated lipid peroxidation rather than protein oxidation. Moreover, Q prevented glycated A1c formation, while no protective effect on the endogenous antioxidant system (GSH-GSSG) was observed. These findings suggest that the B3p could be a novel potential target of antioxidant treatments to counteract aging-related disturbances. Further studies are needed to confirm the possible role of Q in pharmacological strategies against aging.

Keywords: aging; band 3 protein function; d-Galactose; glycation; oxidative stress; quercetin.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1

Figure 1

Chemical structure of Quercetin.

Figure 2

Figure 2

TBARS levels measured in erythrocytes left untreated or after treatment with increasing concentrations (10 µM, 50 µM, or 1 mM) of Q or 10 mM H2O2 (positive control) with or without pre-incubation with Q. ns, not statistically significant versus untreated (control); ***, p < 0.001 versus control; °°°, p < 0.001 versus 10 mM H2O2, one-way ANOVA followed by Bonferroni’s post hoc test (n = 10).

Figure 3

Figure 3

Time course of SO42− uptake in erythrocytes left untreated (control) or treated with 10 µM Q, 100 mM

d

-Gal with or without pre-exposure to 10 µM Q, or 10 µM DIDS. ns, not statistically significant versus control; ***, p < 0.001 versus control; °°°, p < 0.001 versus 100 mM

d

-Gal-treated erythrocytes, one-way ANOVA followed by Bonferroni’s post hoc test.

Figure 4

Figure 4

TBARS levels (µM) in untreated erythrocytes (control) and in erythrocytes treated for 24 h with 100 mM d-Gal, or, alternatively, with 10 µM Q (pre-incubation for 1 h) with or without d-Gal. ns, not statistically significant versus untreated; ***, p < 0.001 significant versus control; °°°, p < 0.001 versus 100 mM

d

-Gal, one-way ANOVA followed by Bonferroni’s post hoc test (n = 5).

Figure 5

Figure 5

Sulfhydryl group content (µM TNB/µg protein) in untreated erythrocytes (control) and in erythrocytes treated with 100 mM

d

-Gal for 24 h with or without 10 µM Q (pre-incubation for 1 h), or alternatively, with 2 mM NEM or 10 µM Q for 1 h. ns, not statistically significant versus control; **, p < 0.01 and ***, p < 0.001 versus control; °°°, p < 0.001 versus 100 mM

d

-Gal, one-way ANOVA followed by Bonferroni’s multiple comparison post hoc test (n = 6).

Figure 6

Figure 6

Glycated hemoglobin content (%A1c) in erythrocytes left untreated or incubated for 24 h with 100 mM

d

-Gal with or without pre-exposure to 10 µM Q, or 10 µM Q alone. ns, not statistically significant versus untreated; ***, p < 0.001 versus untreated; °°°, p < 0.001 versus 100 mM

d

-Gal, one-way ANOVA with Bonferroni’s multiple comparison post hoc test (n = 8).

Figure 7

Figure 7

Intracellular GSH/GSSG ratio measured in erythrocytes left untreated or incubated for 24 h with 100 mM

d

-Gal with or without pre-treatment with 10 µM Q or 10 µM Q alone. ns, not statistically significant versus untreated; ***, p < 0.001 versus untreated (control); °°, p < 0.01 versus 100 mM

d

-Gal, one-way ANOVA followed by Bonferroni’s multiple comparison post hoc test (n = 7).

Figure 8

Figure 8

Time course of experimental procedures before SO42− uptake measurement.

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