Synergistic Effects of a Novel Combination of Natural Compounds Prevent H2O2-Induced Oxidative Stress in Red Blood Cells

Novel strategies to prevent the "storage lesions" of red blood cells (RBCs) are needed to prevent the risk of adverse effects after blood transfusion. One option could be the supplementation of stored blood bags with natural compounds that may increase the basal load of antioxidant protection and the shelf life of RBCs. In this pilot study, we investigated for the first time potential synergistic effects of a triple combination of well-known anti-oxidant compounds curcumin (curc), vitamin E (vit E), and vitamin C (vit C). Briefly, we established an ex vivo model of H₂O₂-induced oxidative stress and measured the hemolysis ratio (HR) (%) and thiobarbituric acid reactive substances (TBARS) levels in RBCs with or without pre-exposure for 30 min with increasing concentrations of curc, vit E, and vit C and then exposed to 10 mM H₂O_2. for 60 min. Exposure of RBCs to a triple combination of curc, vit E, and vit C at the highest concentration (100 µM) completely prevented H₂O₂-induced hemolysis. Surprisingly, we found that pre-treatment of RBCs with curc 100 µM alone completely prevented hemolysis as compared to vit E and vit C alone or in combination at the same concentration. On the other hand, pre-treatment with the triple combination of curc, vit E, and vit C 100 µM was required to totally prevent lipid peroxidation, as compared to curc 100 µM alone, supporting their synergistic effects in preventing RBCs membrane peroxidation. Further experiments are ongoing to investigate the anti-aging effects of the triple combination of curc, vit E, and vit C on cold-stored bags.