title: Vitamin C and Trolox decrease oxidative stress and hemolysis in cold-stored human red blood cells.
abstract: OBJECTIVE
  To investigate the effects of sodium ascorbate (SA) (5-3125 μM) and a combination of SA and Trolox (25 and 125 μM) on oxidative changes generated in red blood cells (RBCs) followed by up to 20 days refrigerated storage.
  
  
  METHODS
  RBCs were isolated from CPD-preserved human blood. Percentage of hemolysis and extracellular activity of lactate dehydrogenase (LDH) were measured to assess the RBC membrane integrity. Lipid peroxidation (LPO), glutathione (GSH) and total antioxidant capacity (TAC) were quantified by thiobarbituric acid-reactive substances, Ellman's reagent and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate) [Formula: see text]-based assay, respectively.
  
  
  RESULTS
  SA failed to reduce the storage-induced hemolysis and RBC membrane permeability. Addition of SA resulted in a concentration-independent LPO inhibition and increased TAC. A combination of SA/Trolox supplemented to the RBC medium significantly inhibited hemolysis, LDH leakage, LPO, GSH depletion and enhanced TAC.
  
  
  CONCLUSIONS
  The effects of vitamin C action are closely concentration-dependent and may be modulated by a variety of compounds (e.g. Hb degradation products) released from RBCs during the prolonged storage, changing its properties from anti- to pro-oxidative. The two different class antioxidants (SA/Trolox) could possibly cooperate to be good potential RBC storage additives ensuring both antiradical and membrane stabilizing protection.
authors.affiliation: a Department of General Biochemistry, Faculty of Biology and Environmental Protection , University of Lodz , Lodz , Poland.
authors.forename: Kamila
authors.initials: K
authors.lastname: Czubak
authors.affiliation: a Department of General Biochemistry, Faculty of Biology and Environmental Protection , University of Lodz , Lodz , Poland.
authors.forename: Adam
authors.initials: A
authors.lastname: Antosik
authors.affiliation: a Department of General Biochemistry, Faculty of Biology and Environmental Protection , University of Lodz , Lodz , Poland.
authors.forename: Natalia
authors.initials: N
authors.lastname: Cichon
authors.affiliation: a Department of General Biochemistry, Faculty of Biology and Environmental Protection , University of Lodz , Lodz , Poland.
authors.forename: Halina Malgorzata
authors.initials: HM
authors.lastname: Zbikowska
mesh_terms: D001205:Ascorbic Acid / Q000494:pharmacology*; D002478:Cells, Cultured; D002839:Chromans / Q000494:pharmacology*; D004912:Erythrocytes / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D011309:Preservation, Biological
chemical_list: D002839:Chromans; D007770:L-Lactate Dehydrogenase; D005978:Glutathione; D001205:Ascorbic Acid; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid
keywords: Oxidative damage; Trolox; red blood cell; sodium ascorbate (vitamin C); storage