Protective Effect of Natural and Synthetic Anthocyanins against Tert-butyl-hydroperoxide-induced Oxidative Damages in Normal and β-thalassemic Major Human Erythrocytes In Vitro

Abstract

Background: Even though β-thalassemia major is a genetic blood disorder, the damages endured by erythrocytes are mediated in part by oxidative stress. Antioxidants such as anthocyanins are capable to prevent the pro-oxidant effects induced by reactive oxygen species (ROS). Objective: This study aims to evaluate the in vitro preventive effects of one natural and two synthetic anthocyanins on normal and β-thalassemic erythrocytes on which toxicity has been induced by the free radical generator: tert-butyl-hydroperoxide TBHP. Methods: Erythrocytes isolated from fasting blood samples of healthy and β-thalassemic major individuals were treated either with TBHP alone or with TBHP after being pre-incubated with anthocyanins. Cell viability, reduced glutathione (GSH) and malondialdehyde (MDA) contents were measured after 90 minutes of incubation. In parallel, the antiradical scavenging capacities of the investigated anthocyanins were also estimated by using the 2,2-DiPhenyl-1-PicrylHydrazyl (DPPH•) assay. Results: The results clearly demonstrate that the treatment of erythrocytes with TBHP induces hemolysis along with marked redox state alteration (lipid peroxidation concomitant to GSH depletion) in both normal and β-thalassemia erythrocytes. During the pre-treatment with anthocyanins, erythrocytes become more resistant to oxidative impairments. Cyanin chloride and 6,7,3’,4’- tetrahydroxyflavylium chloride effectively prevent from TBHP-induced: hemolysis, lipid peroxidation and GSH depletion in normal and thalassemic erythrocytes, while 3’4’-dihdroxy-7-methoxyflavylium chloride had a lesser effect on MDA levels with thalassemic erythrocytes. These results are in agreement with those derived from the DPPH• assay. Conclusion: Our study contributes with important insights that tested anthocyanins may exert relevant potential in the alleviation of oxidative stress, especially the one affecting β-thalassemia erythrocytes.