A comparison of the effect of methyl-$beta$ -cyclodextrin on the osmotic fragility of ovine, bovine and human erythrocytes

Siklodekstrinler gıda, kozmetik, ilaç ve kimya sanayiinde kullanım alanı bulan bazı etken maddelerin çözünürlüğünü ve stabilitesini artırmak, etken maddelerde bulunan bazı istenmeyen tat ve kokuları maskelemek için yaygın olarak kullanılmaktadır. Bu çalışmada pre-hemolitik dozda kullanılan metil-ß-siklodekstrinin (MBCD) koyun, inek ve insan eritrositi ozmotik frajilitesi üzerine etkisi araştırıldı.Bu üç türden elde edilen heparinli kan, alımı takiben üç saat içinde analiz edildi. Eritrositlerden 2, 4 veya 6 mM MBCD içerecek şekilde süspansiyonlar hazırlandı. Karışım 37 $circ$C ısıda 30 dk inkübe edildikten sonra santrifüj edildi. Tekrar sulandırılan eritrositlerin ozmotik frajiliteleri ölçüldü. Her üç türde de eritrosit süspansiyonu ile inkübe edilen MBCD, eritrositlerin ozmotik frajilitesinde doza paralel olarak artan bir ozmotik frajiliteye yol açtı. Hemoliz başlangıcı koyun, inek ve insan kontrol gruplarında sırası ile %0,85, %0,70 ve %0,55 NaCl olarak bulundu. Bunun aksine MBCD uygulanan grupların hepsinde de hemoliz, bundan çok daha erken başladı. Eritrositlerin ozmotik frajilitesi NaCl solüsyonunun %0.70 (P>0.01), %0.75 (P>0.001) ve %0.80 (P>0.001) konsantrasyonlarında koyunda, %0.55 (P>0.01), %0.60 (P>0.001) ve %0.65 (P>0.01) konsantrasyonlarında inekte ve %0.40 (P>0.01), %0.45 (P>0.001) ve %0.50 (P>0.01) konsantrasyonlarında ise insanda kontrol gruplarınkinden daha fazla bulundu. Sonuç olarak; pre-hemolitik dozda kullanılan siklodextrine bağlı olarak eritrosit frajilitesinde meydana gelen artış, bu makromolekülün düşük dozlarının dahi membran komponentlerini uzaklaştırarak, eritrosit membran dayanıklılığını azalttığına işaret etmektedir.

İnek, Koyun ve insaneritrositi ozmotik fajilitesi üzerine metil-$beta$ -siklodextrinin karşılaştırmalı etkisi

Cyclodextrins are used extensively in food, cosmetic, drug and chemical industries to increase aqueous solubility and stability, and reduce or eliminate the unpleasant taste and smell of many products. Experiments were conducted to examine the effect of a pre-haemolytic dose of methyl ß-cyclodextrin (MBCD) on the osmotic fragility of erythrocytes collected from sheep, cattle and humans.Heparinised blood collected from these three species were analysed within 3 h of collection. Erythrocyte suspensions were mixed with MBCD to give concentrations of 2 mM, 4 mM or 6 mM cyclodextrin. The mixtures were incubated for 30 min at 37 $circ$C and centrifuged. The osmotic fragility of re-suspended erythrocytes was measured.The incubation of MBCD with the erythrocyte suspensions induced a dose-dependent increase in the osmotic fragility of erythrocytes obtained from all three species. The beginning of haemolysis in control groups of sheep, cattle and humans occurred at 0.85%, 0.70% and 0.55% NaCl, respectively. However, haemolysis in the MBCD-treated groups began earlier than this in all the three species studied. The osmotic fragility of erythrocytes was significantly higher at NaCl concentrations of 0.70% (P > 0.01), 0.75% (P > 0.001) and 0.80% (P > 0.001) in sheep; 0.55% (P > 0.01), 0.60% (P > 0.001) and 0.65% (P > 0.01) in cattle; and 0.40% (P > 0.01), 0.45% (P > 0.001) and 0.50% (P > 0.01) in humans than in the controls. In conclusion, these results indicate that a pre-haemolytic dose of MBCD may also induce membrane disruption which elicits removal of membrane components from erythrocytes.

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Pitha, J.: Enhanced water solubility of vitamins A, D, E, and K by substituted cycloamyloses. Life Sci. 1981; 29: 307-311.
Pagington, J.S.: |3-cyclodextrins: the success of molecular inclusion. Chemistry in Britain. 1987; 23: 455-458
Pszczola, D.E.: Production and potential food applications of cyclodextrins. Food Techno!.-Chicago 1988; 42: 96-100.
Albers, E., Muller, B.W.: Complexation of steroid hormones with cyclodextrin derivatives: substituent effects of the guest molecule on solubility and stability in aqueous solution. J. Pharm. Sci. 1992; 81: 756-761.
Irie, T., Fukunaga, K., Pitha, J.: Hydroxypropylcyclodextrins in parenteral use. 1: Lipid dissolution and effects on lipid transverse in vitro. J. Pharm. Sci. 1992; 81: 521-523.
Uekama, K., Irie, T.: Cyclodextrins and their industrial uses. In Pharmaceutical applications of methylated cyclodextrin derivatives. Edited by Duchene, D. Editions de Sante, Paris, 1987; 395-439.
Loftsson, T., Brewster, M.E.: Pharmaceutical applications of cyclodextrins. 1. Drug solubilization and stabilisation. J. Pharm. Sci. 1996; 85: 1017-1025.
Kritharides, L.M., Kus, A.J., Brown, W.J., Dean, J.T.: Hydroxypropyl-S-cyclodextrins-mediated efflux of 7- ketocholesterol from macrophage foam cells. J. Biol. Chem. 1996; 271: 27,450-27,455.
Haynes, M.P., Phillips, M.C., Pool, G.H.: Efflux of cholesterol from different cellular pools. Biochemistry. 2000; 39: 4508-4517.
Ohtani, Y., Irie, T., Uekama, K., Fukunaga, K., Pitha, J.: Differential effects of a-,|3- and y-cyclodextrins on human erythrocytes. Eur. J. Biochem. 1989; 186: 17-22.
Kilsdonk, E.P.C,, Yancey, P.G., Stoudt, G.W., Bangerter, F.W., Johnson, W.J., Phillips, M.C.. Rothblat, G.H.: Cellular cholesterol efflux, mediated by cyclodextrin. J. Biol. Chem. 1995; 270: 17,250-17,256.
Christian, A.E., Haynes, M.P. Phillips, M.C., Rothblat, G.H.: Use of cyclodextrins for manipulating cellular cholesterol content. J. Lipid Res. 1997; 38: 2264-2272
Yancey, P.G., Rodrigueza, W.V., Kilsdonk, E.P.C, Stoudt, G.W., Johnson, W.J., Phillips, M.C.. Rothblat, G.H.: Cellular cholesterol efflux mediated by cyclodextrins: demonstration of kinetic pools and mechanism of efflux. J. Biol. Chem. 1996; 271: 16,026- 16,034.
Neufeld, E.B., Cooney, A.M., Pitha, J., Dawidowicz, E.A., Dwyer N.K., Pentchev, P.G., Mackie, E.J.B.: Intracellular trafficking of cholesterol monitored with a cyclodextrin. J. Biochem: Chem. 1996:271:21604-21613.
Voet, D., Voet, J.G.: Lipids and Membranes. In Biochemistry. 2nd edition. John Wiley and Sons, Inc. New York. USA. 1995; p. 290.
Irie, T., Otagiri, M., Sunada, M., Uekama, K., Ohtani, Y., Yamada, Y., Sugiyama, Y.: Cyclodextrin-induced hemolysis and shape changes of human erythrocytes in vitro. J. Pharmacobiodyn. 1982; 5: 741-744.
Shiotani, K., Uehata, K., Irie, T., Uekama, K., Thomson, D.O., Stella, V.J.: Differential effects of sulphate and sulfobutyl ether of |3-cyclodextrins on erythrocyte membranes in vitro. Pharm. Res. 1995; 12: 78-84
Perk, K., Frei, Y.F., Herz, A.: Osmotic fragility of red blood cells of young and mature domestic and laboratory animals. Am. J. Vet. Res. 1964; 25: 1241-1248.
Soliman, M.K., Amrousi, S.E.: Erythrocyte fragility of healthy fowl, dog, sheep, cattle, buffolo, horse, and camel blood. Vet. Rec. 1966; 78: 429-430.
Jikuya, T., Tsutsui, T., Shigeta, 0., Sankai, Y., Mitsui, T.: Species differences in erythrocytemechanical fragility: comparison of human, bovine and ovine cells. ASAIO. J. 1998; 44: M452-455.
Schalm, O.W., Jain, N.C., Carroll, E.S.: Materials and methods for study of the blood. In Veterinary Hematology. 3rd edition. Lea and Febiger, Philadelphia. 1975; 122-151.
Guyton, A.C.: Red blood cells, anemia and polycythemia. In Textbook of medical physiology. 7th edition. W.B. Saunders Company, London. 1986; 43-50.
Parpart, A.K., Loreriz. P.B., Parpart, E.R., Gregg, J.R., Chase, A.M.: The osmotic resistance (fragility) of human red cells. J. Clin. Invest. 1947; 26: 636-643.
Jodal, I., Nanasi, P., Szejtli, J.: Investigation of the haemolytic effect of the cyclodextrin derives. In Advances in inclusion science, Proceedings of the Fourth International Symposium on Cyclodextrins, Munich, West Germany. Edited by Huber, 0., Szejtli, J. Kluwer Academic Publishers, Boston. April 20-22, 1988:421-425.
Macarak, E.J., Kumor, K., Weisz, P.B.: Sulfation and haemolytic activity of cyclodextrin. Biochem. Pharm. 1991; 42: 1502-1503. 26.Greatorex, J.C.: Observation on the haematology of calves and various breeds of adult dairy cattle. Brit. Vet. J. 1957; 113: 469- 479.
Atger, V.M., Moya, M.L, Stoudt, G.W., Rodrigueza, W.V., Phillips, M.C., Rothblat, G.H.: Cyclodextrins as catalysts for the removal of cholesterol from macrophage foam cells. J. Clin. Invest. 1997; 99: 773-780.