Supplementation with uric and ascorbic acid protects stored red blood cells through enhancement of non-enzymatic antioxidant activity and metabolic rewiring

Vassilis L Tzounakas; Alkmini T Anastasiadi; Vasiliki-Zoi Arvaniti; Veronica Lelli; Giuseppina Fanelli; Efthymios C Paronis; Anastasia C Apostolidou; Evangelos G Balafas; Nikolaos G Kostomitsopoulos; Effie G Papageorgiou; Issidora S Papassideri; Konstantinos Stamoulis; Anastasios G Kriebardis; Sara Rinalducci; Marianna H Antonelou

doi:10.1016/j.redox.2022.102477

Supplementation with uric and ascorbic acid protects stored red blood cells through enhancement of non-enzymatic antioxidant activity and metabolic rewiring

Redox Biol. 2022 Nov:57:102477. doi: 10.1016/j.redox.2022.102477. Epub 2022 Sep 20.

Authors

Vassilis L Tzounakas¹, Alkmini T Anastasiadi¹, Vasiliki-Zoi Arvaniti¹, Veronica Lelli², Giuseppina Fanelli², Efthymios C Paronis³, Anastasia C Apostolidou³, Evangelos G Balafas³, Nikolaos G Kostomitsopoulos³, Effie G Papageorgiou⁴, Issidora S Papassideri¹, Konstantinos Stamoulis⁵, Anastasios G Kriebardis⁴, Sara Rinalducci⁶, Marianna H Antonelou⁷

Affiliations

¹ Department of Biology, School of Science, National and Kapodistrian University of Athens (NKUA), Athens, Greece.
² Department of Ecological and Biological Sciences, University of Tuscia, Viterbo, Italy.
³ Center of Clinical, Experimental Surgery & Translational Research, Biomedical Research Foundation, Academy of Athens (BRFAA), Athens, Greece.
⁴ Laboratory of Reliability and Quality Control in Laboratory Hematology (HemQcR), Department of Biomedical Sciences, School of Health & Welfare Sciences, University of West Attica (UniWA), Egaleo, Greece.
⁵ Hellenic National Blood Transfusion Centre, Acharnes, Greece.
⁶ Department of Ecological and Biological Sciences, University of Tuscia, Viterbo, Italy. Electronic address: sara.r@unitus.it.
⁷ Department of Biology, School of Science, National and Kapodistrian University of Athens (NKUA), Athens, Greece. Electronic address: manton@biol.uoa.gr.

Abstract

Redox imbalance and oxidative stress have emerged as generative causes of the structural and functional degradation of red blood cells (RBC) that happens during their hypothermic storage at blood banks. The aim of the present study was to examine whether the antioxidant enhancement of stored RBC units following uric (UA) and/or ascorbic acid (AA) supplementation can improve their storability as well as post-transfusion phenotypes and recovery by using in vitro and animal models, respectively. For this purpose, 34 leukoreduced CPD/SAGM RBC units were aseptically split in 4 satellite units each. UA, AA or their mixture were added in the three of them, while the fourth was used as control. Hemolysis as well as redox and metabolic parameters were studied in RBC units throughout storage. The addition of antioxidants maintained the quality parameters of stored RBCs, (e.g., hemolysis, calcium homeostasis) and furthermore, shielded them against oxidative defects by boosting extracellular and intracellular (e.g., reduced glutathione; GSH) antioxidant powers. Higher levels of GSH seemed to be obtained through distinct metabolic rewiring in the modified units: methionine-cysteine metabolism in UA samples and glutamine production in the other two groups. Oxidatively-induced hemolysis, reactive oxygen species accumulation and membrane lipid peroxidation were lower in all modifications compared to controls. Moreover, denatured/oxidized Hb binding to the membrane was minor, especially in the AA and mix treatments during middle storage. The treated RBC were able to cope against pro-oxidant triggers when found in a recipient mimicking environment in vitro, and retain control levels of 24h recovery in mice circulation. The currently presented study provides (a) a detailed picture of the effect of UA/AA administration upon stored RBCs and (b) insight into the differential metabolic rewiring when distinct antioxidant "enhancers" are used.

Keywords: Antioxidant supplementation; Ascorbic acid; Glutathione metabolism; Oxidative stress; RBC storage lesion; Uric acid.