Structure-activity relationships regarding the antioxidant effects of the flavonoids on human erythrocytes


The effects of eleven flavonoids on lipid peroxidation, protein degradation, deformability and osmotic fragility of human erythrocytes exposed in vitro to 10 mM H2O2 for 60 min at 37 oC have been studied. The following flavonoids; quercetin, rutin and morin significantly protected eryt-hrocytes against lipid peroxidation caused by H2O2. This inhibition of lipid peroxidation could be explained by the presence of at least two hydroxyl groups in ring B of the flavonoid structure, regardless of their positions. However, the flavonoids; quercetin, 3,5,7-trihy- droxy-4'-methoxy flavone-7-rutinoside and 3- hydroxy flavone significantly protected eryt-hrocytes against protein degradation. This inhibition could also be explained by the presence of a hydroxyl group at C-3 in ring C of the flavonoid structure. Quercetin and 3,5,7-trihydroxy-4'- methoxy flvone-7-rutinoside significantly protected erythrocytes against loss of deformability and increased osmotic fragility, indicating that the loss of erythrocyte deformability and the increase in osmotic fragility of erythrocytes exposed to H2O2 are related to protein degradation rather than to lipid peroxidation. The other flavonoids (chrysin, 2-carboxy ethyl dihydroxy flavone, apigenin, cirsimaritin, α-naphto flavone and flavanone) failed to protect erythrocytes against the observed oxidative damages. The results demonstrate the importance of the chemical groups substituted on the basic skeleton of the flavonoids in dictating the type of antioxidant activity, and also demonstrate the hemorheological potentials of flavonoids that have particular protein-antioxidant activities.

Share and Cite:

Bilto, Y. , Suboh, S. , Aburjai, T. and Abdalla, S. (2012) Structure-activity relationships regarding the antioxidant effects of the flavonoids on human erythrocytes. Natural Science, 4, 740-747. doi: 10.4236/ns.2012.49098.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Chiu, D., Kuypers, E. and Lubin, B. (1989) Lipid peroxidation in human red cells. Hematology, 26, 257-276.
[2] Davies, K.J.A. and Goldberg, A.L. (1987) Oxygen radicals stimulate intracellular proteolysis and lipid peroxidetion by independent mechanisms in erythrocytes. The Journal of Biological Chemistry, 262, 8220-8226.
[3] Saltman, P. (1989) Oxidative stress: A radical view. Hematology, 26, 249-256.
[4] Srour, M.A., Bilto, Y.Y. and Juma, M. (2000) Susceptibility of erythrocytes from non-insulin-dependent diabetes mellitus and hemodialysis patients, cigarette smokers and normal subjects to in vitro oxidative stress and loss of deformability. Microvascular Biology and Inflammation, 22, 173-180.
[5] Trotta, R.G., Sullivan, S.G. and Stern, A. (1981) Lipid peroxidation and hemoglobin degradation in red blood cells exposed to t-butyl hydroperoxide. Biochimica et Biophysica Acta, 679, 230-237. Hdoi:10.1016/0304-4165(81)90211-7
[6] Snyder, L.M., Fortier, N.L., Trainor, J., Jacobs, J., Leb, L., Lubin, B., Chiu, D., Shohet, S. and Mohandas N. (1985) Effect of hydrogen peroxide exposure on normal human erythrocyte deformability, morphology, surface characteristics and spectrin-hemoglobin cross-linking. Journal of Clinical Investigation, 76, 1971-1977. Hdoi:10.1172/JCI112196
[7] Chiu, D. and lubin, B. (1989) Oxidative hemoglobin denaturation and RBC destruction: The effect of heme on red cell membranes. Hematology, 26, 128-135.
[8] Van der zee, J., Steveninck, J.V., Koster, J.V., Koster, J.F. and Dubbel, Man, T.M. (1989) Inhibition of enzymes and oxidative damage of red blood cells induced by t-butyl hydroperoxide-derived radicals. Biochimica et Biophysica Acta, 980, 175-180.
[9] Pradhan, D., Weiser, M., Lumley-Sapanski, K., Frazier, D., Williamson, P. and Schlegel, R.A. (1990) Peroxidation induced perturbations of erythrocyte lipid organization. Biochimica et Biophysica Acta, 1023, 398-404. Hdoi:10.1016/0005-2736(90)90132-8
[10] Van den Berg, J.J., Op den Kamp, J.A., Lubin, B.H., Roelofsen, B. and Kuypers, F.A. (1992) Kinetics and site specificity of hydroperoxide-induced oxidative damage in red blood cells. Free Radical Biology & Medicine, 12, 487-498. Hdoi:10.1016/0891-5849(92)90102-M
[11] Sato, Y., Kamo, S., Takahashi, T. and Suzuki, Y. (1995) Mechanism of free radical-induced hemolysis of human erythrocytes: Hemolysis by water-soluble radical initiator. Biochemistry, 34, 8940-8949. Hdoi:10.1021/bi00028a002
[12] Srour, M.A., Bilto, Y.Y., Juma, M. and Irhimeh, M.R. (2000) Exposure of human erythrocytes to oxygen radicals causes loss of deformability, increased osmotic fragility, lipid peroxidation and protein degradation. Clinical Hemorheology and Microcircul, 23, 1-9.
[13] Halliwell, B. (1989) Protection against tissue damage in vivo by desferrioxamine: What is its mechanism of action? Free Radical Biology and Medicine, 7, 645-651. Hdoi:10.1016/0891-5849(89)90145-7
[14] Miller, A.L. (1996). Antioxidant flavonoids: Structure, function and clinical usage. Alternative Medicine Review, 1, 103.
[15] Cao, G. Sofic, E.R. and Prior, R.L. (1996) Antioxidant capacity of tea and common vegetables. Journal of Agricultural and Food Chemistry, 44, 3426-3431. Hdoi:10.1021/jf9602535
[16] Block, G. and Patterson, B. (1992) Fruits, vegetables and cancer prevention: A review of the epidemiological evidence. Nutrition and Cancer, 18, 1-29. Hdoi:10.1080/01635589209514201
[17] Ness, A.R. and Powles, J.W. (1997) Fruit and vegetables and cardiovascular disease: A review. International Journal of Epidemiology, 26, 1-13. Hdoi:10.1093/ije/26.1.1
[18] Cai, Y. Luo, Q. Sun, M. and Corke, H. (2004) Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer. Life Science, 74, 2157-2184. Hdoi:10.1016/j.lfs.2003.09.047
[19] Tawaha, F., Gharaibeh, M., Mohammad, M. and El-Elimate, T. (2007) Antioxidant activity and total phenolic content of selected Jordanian plant species. Food Chemistry, 104, 1372-1378. Hdoi:10.1016/j.foodchem.2007.01.064
[20] Rice-Evans, C.A., Miller, N.J. and Paganga, G. (1996) Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Radical Biology and Medicine, 20, 933-956. Hdoi:10.1016/0891-5849(95)02227-9
[21] Rice-Evans, C.A. Miller, N.J. and Paganga, G. (1997) Antioxidant properties of phenolic compounds. Trends in Food Science & Technology, 2, 152-159.
[22] Harborne, J.B., (1967) Comparative biochemistry of flavonoids. Academic Press Inc., London.
[23] Hall, III, C.A. and Cuppett, S.L. (1997) Structure-activeties of natural antioxidants. In: Aruoma, O.I. and Cuppett, S.L., Eds., Antioxidant methodology: In vivo and in vitro concepts. AOCS Press, Illinois, 141-172.
[24] Suboh, S.M., Bilto, Y.Y. and Aburjai, T.A. (2004) Protective effects of selected medicinal plants against protein degradation, lipid peroxidation and deformability loss of oxidatively stressed human erythrocytes. Phytotherapy Research, 18, 280-284. Hdoi:10.1002/ptr.1380
[25] Dacie, J.V. and Lewis, S.M. (1995) Practical haematology. 8th Edition, Churchil Livingstone, Edinburgh.
[26] Stocks, J. and Dormandy, T.L. (1971) The autoxidation of human red cell lipids induced by hydrogen peroxide. British Journal of Haematology, 20, 95-111. Hdoi:10.1111/j.1365-2141.1971.tb00790.x
[27] Srour, M.A., Bilto, Y.Y. and Juma, M. (2000C) Evaluation of different methods used to measure malonyldialdehyde in human erythrocytes. Clinical Hemorheology and Microcircul, 23, 23-30.
[28] Davies, K.J.A. and Goldberg, A.L. (1987) Proteins damaged by oxygen radicals are rapidly degraded in extracts of red blood cells. The Journal of Biological Chemistry, 262, 8227-8234.
[29] Bilto, Y.Y., Player, M., West, M.J., Ellory, J.C. and Stuart, J. (1987) Effects of oxpentifylline on erythrocyte cation content, hydration and deformability. Clinical Hemorheology, 7, 561-577.
[30] Hanss M. (1983) Erythrocyte filterability measurement by the initial flow rate method. Biorheology, 20, 199-211.
[31] Bilto, Y.Y. and Stuart, J. (1985) Ultrasonic cleaning of polycarbonate membranes for measurement of erythrocyte filterability. Clinical Hemorheology, 5, 434-448.
[32] Luqman, S., Kaushik, S., et al. (2009) Protective effect of medicinal plant extracts on biomarkers of oxidative stress in erythrocytes. Pharmaceutical Biology (Formerly International Journal of Pharmacognosy), 47, 483-490. Hdoi:10.1080/13880200902832900
[33] Brown, J.E., khodr, H., Hider, R.C. and Rice-Evans, C.A. (1998) Structural dependence of flavonoid interactions with Cu2+ ions: Implications for their antioxidant properties. Biochemical Journal, 330, 1173-1178.
[34] Choi, J.S., et al. (2002) The structure—Activity relationship of flavonoids as scavengers of peroxynitrite. Phytotherapy Research, 16, 232-235. Hdoi:10.1002/ptr.828
[35] Ferrali, M., Signorini, C., Caciotti, B., Sugherini, L., Ciccoli, L., Giachetti, D. and Comporti, M. (1997) Protection against oxidative damage of erythrocyte membrane by the flavonoid quercetin and its relation to iron chelating activity. FEBS letters, 416, 123-129. Hdoi:10.1016/S0014-5793(97)01182-4
[36] Murakami, S., Muramatsu, M. and Tomisawa, K. (1999) Inhibition of gastric H+, K+-ATPase by flavonoids: A structure-activity study. Journal of Enzyme Inhibition, 14, 151-166. Hdoi:10.3109/14756369909036551
[37] Pietta, P.G. (2000) Flavonoids as antioxidants. Journal of Natural Products, 63, 1036-1042. Hdoi:10.1021/np9904509
[38] De Beer, D., Joubert, E., Gelderblom, W.C.A. and Manley, M. (2002) Phenolic compounds: A review of their possible role as in vivo antioxidants of wine. South African Journal for Enology and Viticulture, 23, 48-61.
[39] Perron, N.R. and Brumaghim, J.L. (2009) A review of the antioxidant mechanisms of polyphenol compounds related to iron binding. Cell Biochemistry and Biophysics, 53, 75-100. Hdoi:10.1007/s12013-009-9043-x
[40] Rice-Evans, C. (2001) Flavonoid antioxidants. Current Medicinal Chemistry, 8, 797-807.
[41] Pathak, D., Pathak, K. and Singla, A.K. (1991) Flavanoids as medicinal agents-recent advances. Fitoterapia, 62, 371-389.
[42] Affany, A., Salvayre, R. and Douste-Blazy, L. (1987) Comparison of the protective effect of various flavonoids against lipid peroxidation of erythrocyte membranes (induced by cumene hydroperoxide). Fundamental & Clinical Pharmacology, 1, 451-457. Hdoi:10.1111/j.1472-8206.1987.tb00578.x
[43] Ruchstuhl, M. and Landry, Y. (1981) Inhibition of lung cyclic AMP- and cyclic GMP-phosphodiesterases by flavonoids and other chromone-like compounds. Biochemical Pharmacology, 30, 697-702. Hdoi:10.1016/0006-2952(81)90153-2
[44] Foti, M., Piatelli, Baratta, M.T. and Ruberto, G. (1996) Flavonoids, coumarins, and cinnamic acids as antioxidants in a micellar system. Structure-activity relationship. Journal of Agricultural and Food Chemistry, 44, 497- 501. Hdoi:10.1021/jf950378u
[45] Liao, K.-L. and Yin, M.-C. (2000) Individual and combined antioxidant effects of seven phenolic agents in human erythrocyte membrane ghosts and phosphatidylrcholine liposome systems: Importance of the partition coefficient. Journal of Agricultural and Food Chemistry, 48, 2266-2270. Hdoi:10.1021/jf990946w
[46] Abu-Khader, A.A. and Bilto, Y.Y. (2002) Exposure of human neutrophils to oxygen radicals causes loss of deformability, lipid peroxidation, protein degradation, respiratory burst activation and loss of migration. Clinical Hemorheology and Microcircul, 27, 57-66.

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.