Vitamin C with N-acetylcysteine ameliorates the antioxidant defenses in erythrocytes during storage

Yıl 2024, Cilt: 41 Sayı: 2, 302 - 310, 19.05.2024

Masannagari Pallavi Vani Rajashekaraiah

Öz

Oxidative stress causes structural and biochemical alterations in stored erythrocytes. Antioxidants can be employed to prevent oxidative damage and enhance antioxidant defenses, thereby improving the efficacy of stored erythrocytes. N-acetylcysteine (a glutathione precursor), and Vitamin C (a free radical scavenger) can be potential candidates as additives. Hence, this study investigates the interactions of Vitamin C and N-acetylcysteine during erythrocyte storage. Blood was sampled from Wistar rats, and erythrocytes were grouped into (i) Controls and (ii) Experimentals [vitamin C-10 mM & N-acetylcysteine-0.5 mM]. The erythrocytes were stored in additive solution-7 (AS-7) at 4˚C for a duration of 35 days. Antioxidant and oxidative stress markers were analyzed at weekly intervals. Hemoglobin levels increased on day 14 in Experimentals compared to Controls and normalized on day 35. Catalase activity was maintained in Experimentals, whereas it decreased in Controls. Glutathione peroxidase activity decreased on day 14, whereas it increased on day 35 in Experimentals. Although thiobarbituric acid reactive substances decreased on days 14, 28 & 35 in both groups, there was a steep decline towards the end of storage in Experimentals. Endogenous antioxidant levels were higher throughout storage in Experimentals. Vitamin C with N-acetylcysteine reduced oxidative stress and enhanced antioxidant capacity during storage. This is the first report to elucidate the interactions of vitamin C and N-acetylcysteine in erythrocytes stored in AS-7.

Anahtar Kelimeler

Erythrocytes, Oxidative Stress, Storage, Vitamin C

Etik Beyan

Animal care and maintenance were according to ethical committee regulations (841/b/04/ CPCSEA). Ethical approval was obtained from the Institutional Animal Ethics Committee (IAEC) at the Nargund College of Pharmacy, Bengaluru (IAEC/NCP/117/2022).

Kaynakça

• D’Alessandro A, Liumbruno G, Grazzini G, Zolla L. Red blood cell storage: the story so far. Blood Transfus. 2010;8(8):82-88.
• Bardyn M, Tissot JD, Prudent M. Oxidative stress and antioxidant defenses during blood processing and storage of erythrocyte concentrates. Transfus Clin Bio. 2018; 25(1):96-100.
• Delobel J, Prudent M, Tissot JD, Lion N. Proteomics of the red blood cell carbonylome during blood banking of erythrocyte concentrates. Proteomics Clin Appl. 2016;10(3):257-266.
• Kriebardis AG, Antonelou MH, Stamoulis KE, Economou‐Petersen E, Margaritis LH, Papassideri IS. Progressive oxidation of cytoskeletal proteins and accumulation of denatured hemoglobin in stored red cells. J Cell Mol Med. 2007;11:148-155.
• Mohammed A, Omar A, Christine B, Ibrahim A, Sarra A, Al Ossam Mohammed A. Pathological roles of reactive oxygen species and their defence mechanisms. Saudi Pharmaceut J. 2004;12(1):1-18.
• Soumya R, Vani R. CUPRAC–BCS and antioxidant activity assays as reliable markers of antioxidant capacity in erythrocytes. Hematology. 2015;20(3):165-174.
• Links M, Lewis C. Chemoprotectants: a review of their clinical pharmacology and therapeutic efficacy. Drugs. 1999;57:293-308.
• Moldéus P, Cotgreave IA, Berggren M. Lung protection by a thiol-containing antioxidant: N-acetylcysteine. Respiration. 1986;50:31-42.
• Aruoma OI, Halliwell B, Hoey BM, Butler J. The antioxidant action of N-acetylcysteine: its reaction with hydrogen peroxide, hydroxyl radical, superoxide, and hypochlorous acid. Free Radic Biol Med. 1989;6(6):593-597.
• Wood LA, Beutler E. The effect of ascorbate on the maintenance of 2, 3‐diphosphoglycerate (2,3‐DPG) in stored red cells. British J Hematol.1973;25(5):611-618.
• Raval JS, Fontes J, Banerjee U, Yazer MH, Mank E, Palmer AF. Ascorbic acid improves membrane fragility and decreases haemolysis during red blood cell storage. Transfus Med. 2013;23(2):87-93.
• Stowell SR, Smith NH, Zimring JC, Fu X, Palmer AF, Fontes J, et al. Addition of ascorbic acid solution to stored murine red blood cells increases posttransfusion recovery and decreases microparticles and alloimmunization. Transfusion. 2013;53(10):2248-2257.
• Vani R, Soumya R, Carl H, Chandni VA, Neha K, Pankhuri B, et al. Prospects of vitamin C as an additive in plasma of stored blood. Adv Hematol. 2015;2015: 961049.
• Amen F, Machin A, Tourino C, Rodriguez I, Denicola A, Thomson L. N-acetylcysteine improves the quality of red blood cells stored for transfusion. Arch Biochem Biophys. 2017;621:31-37.
• Soumya R, Vani R. Vitamin C as a modulator of oxidative stress in erythrocytes of stored blood. Acta Haematol Pol. 2017;48(4):350-356.
• Pallotta V, Gevi F, D’Alessandro A, Zolla L. Storing red blood cells with vitamin C and N-acetylcysteine prevents oxidative stress-related lesions: a metabolomics overview. Blood Transfus. 2014;12(3):376-387.
• Jani VP, Yalcin O, Williams AT, Popovsky MA, Cabrales P. Rat red blood cell storage lesions in various additive solutions. Clin hemorheol microcir. 2017;67(1):45-57.
• d'Almeida MS, Jagger J, Duggan M, White M, Ellis C, Chin‐Yee IH. A comparison of biochemical and functional alterations of rat and human erythrocytes stored in CPDA‐1 for 29 days: implications for animal models of transfusion. Transfus Med. 2000;10(4):291-303.
• Pallavi M, Rajashekaraiah V. Synergistic activity of vitamin-C and vitamin-E to ameliorate the efficacy of stored erythrocytes. Transfus Clin Bio. 2023;30(1):87-95.
• da SilveiraCavalcante L, Acker JP, Holovati JL. Differences in rat and human erythrocytes following blood component manufacturing: the effect of additive solutions. Transfus Med Hemother. 2015;42(3):150-157.
• Lagerberg JW, Korsten H, Van Der Meer PF, De Korte D. Prevention of red cell storage lesion: a comparison of five different additive solutions. Blood Transfus. 2017;15(5):456-462.
• Kiran TR, Subramanyam MV, Devi SA. Swim exercise training and adaptations in the antioxidant defense system of myocardium of old rats: relationship to swim intensity and duration. Comp Biochem Physiol B Biochem Mol Biol. 2004;137(2):187-196.
• Dodge JT, Mitchell C, Hanahan DJ. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963;100(1):119-130.
• Hawk FB. Physiological chemistry. New York: Tata McGraw Hill; 1965.
• Misra HP, Fridovich I. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem. 1972;247(10):3170-3175.
• Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121-126.
• Flohe L, Gunzler WA. Assays of glutathione peroxidase. Methods Enzymol. 1984;105:114-120.
• Beutler E, Duran O, Kelley BM. Modified procedure for the estimation of reduced glutathione. J Lab Clin Med. 1963;61:882-888.
• Thomas L editor. Clinical laboratory diagnostics: use and assessment of clinical laboratory results. TH-books Verlagsgesellschaft. 1998
• Olas B, Wachowicz B. Resveratrol and vitamin C as antioxidants in blood platelets. Thromb Res. 2002;106(2):143-148.
• Witko-Sarsat V, Friedlander M, Capeillère-Blandin C, Nguyen-Khoa T, Nguyen AT, Zingraff J, et al. Advanced oxidation protein products as a novel marker of oxidative stress in uremia. Kidney Int. 1996;49(5):1304-13.
• Habeeb AFSA. Reaction of protein sulfhydryl groups with Ellman’s reagent. Methods Enzymol. 1972;25:457-464.
• Bar-Or D, Rael LT, Lau EP, et al. An analog of the human albumin N-terminus (Asp-Ala-His-Lys) prevents formation of copper-induced reactive oxygen species. Biochem Biophys Res Commun. 2001;284(3):856-862.
• Basak A. Development of a rapid and inexpensive plasma glucose estimation by two-point kinetic method based on glucose oxidase-peroxidase enzymes. Indian J Clin Biochem. 2007;22:156-160.
• Lowry OH, Rosenberg NJ, Farr AL, Randall RJ. Protein measurements with Folin-phenol reagent. J Biol Chem. 1951;193:265-75.
• Taylor MB, Christian KG, Patel N, Churchwell KB. Methemoglobinemia: Toxicity of inhaled nitric oxide therapy. Pediatr Crit Care Med. 2001;2(1):99-101.
• Wright RO, Woolf AD, Shannon MW, Magnani B. N‐acetylcysteine reduces methemoglobin in an in‐vitro model of glucose‐6‐phosphate dehydrogenase deficiency. Acad Emerg Med. 1998;5(3):225-229.
• Çimen MB. Free radical metabolism in human erythrocytes. Clin Chimica Acta. 2008;390:1-11.
• Paschalis V, Theodorou AA, Margaritelis NV, Kyparos A, Nikolaidis MG. N-acetylcysteine supplementation increases exercise performance and reduces oxidative stress only in individuals with low levels of glutathione. Free Radic Biol Med. 2018;115:288-97.
• Rosic G, Srejovic I, Zivkovic V, Selakovic D, Joksimovic J, Jakovljevic V. The effects of N-acetylcysteine on cisplatin-induced cardiotoxicity on isolated rat hearts after short-term global ischemia. Toxicol Rep. 2015;2:996-1006.
• Abdallah FB, Gargouri B, Bejaoui H, Lassoued S, Ammar‐Keskes L. Dimethoate‐induced oxidative stress in human erythrocytes and the protective effect of vitamins C and E in vitro. Environ Toxicol. 2011;26(3):287-91.
• Samuni Y, Goldstein S, Dean OM, Berk M. The chemistry and biological activities of N-acetylcysteine. Biochim Biophys Acta. 2013;1830(8):4117-29.
• Pehlivan FE. Vitamin C: An antioxidant agent. Vitamin C. 2017;2:23-35.
• Huyut Z, Şekeroğlu MR, Balahoroğlu R, Huyut MT. Characteristics of resveratrol and serotonin on antioxidant capacity and susceptibility to oxidation of red blood cells in stored human blood in a time-dependent manner. J Int Med Res. 2018;46(1):272-83.
• Carr A, Frei B. Does vitamin C act as a pro‐oxidant under physiological conditions?. The FASEB J. 1999;13(9):1007-1024.
• Eroglu S, Pandir D, Uzun FG, Bas H. Protective role of vitamins C and E in diclorvos-induced oxidative stress in human erythrocytes in vitro. Biol Res. 2013;46(1):33-38.
• Dumaswala UJ, Zhuo L, Mahajan S, Nair PN, Shertzer HG, Dibello P, et al. Glutathione protects chemokine-scavenging and antioxidative defense functions in human RBCs. Am J Physio Cell Physiol. 2001;280(4):867-73.