UVB-induced hemolysis of rat erythrocytes:: Protective effect of procyanidins from grape seeds

• Published in: Life sciences (1973) 2000-09, Vol.67 (15), p.1799-1814
• Main Authors: Carini, M., Aldini, G., Bombardelli, E., Morazzoni, P., Facino, R.Maffei
• Format: Article
• Language: English
• Subjects: Animals; Antioxidants - pharmacology; Biflavonoids; Catechin - analogs & derivatives; Catechin - pharmacology; Chain-breaking antioxidant; Chromatography, High Pressure Liquid; Dose-Response Relationship, Drug; Erythrocyte Membrane - drug effects; Erythrocyte Membrane - metabolism; Erythrocyte Membrane - radiation effects; Erythrocytes - drug effects; Erythrocytes - metabolism; Erythrocytes - radiation effects; Glutathione - metabolism; Hemoglobins - drug effects; Hemoglobins - metabolism; Hemoglobins - radiation effects; Hemolysis - drug effects; Hemolysis - radiation effects; Lipid Peroxidation - drug effects; Lipid Peroxidation - radiation effects; Male; Membrane Fluidity - drug effects; Membrane Fluidity - radiation effects; Oxidants - pharmacology; Oxidation-Reduction; Phospholipids - metabolism; Phospholipids - radiation effects; Phospholipids protection; Proanthocyanidins; Procyanidins from Vitis vinifera; Protective effect; Rat erythrocytes; Rats; Rats, Wistar; Rosales - chemistry; Thiobarbituric Acid Reactive Substances - metabolism; Ultraviolet Rays - adverse effects; UVB-induced hemolysis; Vitamin E - metabolism; Vitamin E saving
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/S0024-3205(00)00771-2

Besides erythema and sunburn reactions, UVB stress can promote erythrocyte extravasation from skin capillaries and hemolysis, and photosensitized hemoglobin can in turn lead to an overload of free radicals in dermis which exacerbates photodamage. The objective of this study was to investigate in rat erythrocytes (RBC) the pattern of events leading to membrane peroxidation and hemolysis following UVB insult (1.5–8.5 J/cm 2), and the protective action of grape seed procyanidins. UVB causes a dramatic dose-dependent decrease of intracellular glutathione (paralleled by the formation of pro-oxidant ferryl-hemoglobin), of intramembrane vitamin E and of membrane fluidity, then a rise of conjugated dienes (CD), and thiobarbituric acid-reactive substances (TBARS) and finally a strong hemolytic effect. Procyanidins prevent membrane peroxidation (but not intracellular GSH depletion nor ferryl-hemoglobin formation), with a minimal effective concentration of 0.1 μM (IC 50 for TBARS and CD after 120 min UVB exposure: 0.71 μM and 0.56 μM) and dose-dependently delay the onset of hemolysis, by 30 min at 0.1 μM, by 90 and 120 min at 0.5 and 1.0 μM. Epigallocatechin-3- O-gallate (EGCG) and catechin, typical constituents of the fraction, were significantly less potent. This since procyanidins (1 μM) inhibit the formation of phospholipid hydroperoxides of the inner (phosphatidylserine, phosphatidylethanolamine) and outer (phosphatidylcholine) layers of the RBC membrane (HPLC analysis), suppress the decrease in membrane fluidity due to lipid and protein thiol oxidation and spare vitamin E from consumption in a dose-dependent manner (0.1–1 μM). Hence procyanidins, preserving membrane phospholipids, since their strong antilipoperoxidant activity, may maintain in vivo the integrity of RBC in sub-epidermal capillaries and effectively counteract in dermis the onset/exacerbation of the UVB-induced skin photodamage.