Clinica Chimica Acra, 203 (1991) 413-416 0 1991 Elsevier Science Publishers B.V. All rights reserved 0009-8981/91/$03.50
413
CCA 05188
Letter
to the Editor
Changes in lipid peroxidation and antioxidant enzyme activity of human red blood cells after myocardial infarction (Received 5 December 1990, revision received 8 October 1991; accepted 15 October 1991)
Dear Editor, If lipid peroxides are increasingly produced in pathological processes the end effect might be thrombosis or cell necrosis [1,2]. Ischemic-reperfusion has been thoroughly studied as a potential source of oxygen radicals. The fundamental importance of oxygen radicals in this processes is also demonstrated in an indirect way by the effectioness of antioxidant therapy [3-71. We have further examined the mechanisms of reperfusion injury and now here to report how lipid peroxidation and antioxidant enzyme activity change 8 h after acute myocardial infarction. Activities of the antioxidant enzymes, superoxide dismutase (SOD; EC 1.15.1.1), catalase (Case; EC 1.11.1.6), glutathione peroxidase (GP-ase; EC 1.11.1.9) and the lipid peroxidation (LPI products were measured in haemolysates of red blood cells (RBCs) harvested from blood taken from the antecubital vein. The methods for these assays were: (i) SOD assay based on the inhibition of epinephrine-adrenochrome transformation [8,9]; (ii) catalase through the rate of H,O, cleavage/unit of time at 240 nm (9,101; (iii) activity of glutathione peroxidase with the help of cumen hydroperoxide co-substrate [ll-141; (iv) LP as total-thiobarbituric acid active material, and expressed as a malondialdehyde (MDA) value, using malondialdehyde diethylacetale for calibration 111,151; (v) protein content by the method of Lowry et al. [16]. Data were analysed for statistical significance with the Student’s t test. The values given are mean f SD. The results are given in Table I. Activities of all the antioxidant enzymes tested decreased. in RBC haemolysates from blood taken 8 h after acute myocardial infarct. While the changes in C-ase and SOD were significant the decrease in GP-ase was not. The LP value showed a pronounced increase [7]. The observed changes in GP-ase activity presented here in the initial period of cardiac infarct similar to those published by Gromadinska et al. 1171.It is intriguing that other observed the increase in GP-ase activity in RBCs after cardiac infarct [18]. It remains a question whether the above mentioned differences may come from the differences in the methods applied in measuring of GP-ase.
414 TABLE I RBC lipid peroxidation and antioxidant (mean f SD1 enzyme activities in myocardial infarction Erythrocyte haemolysate
LP nmol MDA/ml
Myocardial infarct SD *
101.3 2.5.6
2.30 0.61
< 0.01 67.4 15.9
> 0.05 2.57 0.26
group (n = 10) P Control group SD* (n=ll)
GP-ase U/ml haemoi.
SOD U/ml haemol.
Catalase BIJ/ml haemol
493.0 135.0
0.32
< 0.001 716.5 92.6
1.05
< 0.001 2.05 0.68
BU = Bergmeyer unit
Sjgnificant increase in RBC LP after myocardia~ infarct may also have a role in the inhibition of SOD and C-ase activities which results further H,O, accumulation and in a circulus viciosus which triggers again a decrease in enzyme activity. The H,O, accumulation shifting the arachidonic acid metabolism away from the prostacyclin parthway. Aranka L.&lo I A. Gent-Gy6rgyi
‘, B&a Matkovics 2, Sz. Ilona Varge 2, Tibor Wittman 3 and Tamas Fazekas 3 Me&al University, Department of Pediatrics, Szeged,
’ Biological Isotope Labortory, ‘Al ’ University of Szeged and s A. Szent-Gyiirgi Medical U~.~ersi~, 1st De~rt~eat of Internal Medicine, Szeged, Hungary
1 De Maestro RF. An approach to free radicals in medicine and biology. Acta Physiol Stand 1980; Suppl. 492:153-168. 2 McCord J, Fridovich I. The biology and pathology of oxygen radicals. Ann Intern Med 1978;89:122127. 3 Larner AJ, Conway MA. Free radicals in acute myocardial infarction. Quart J Med New Ser 1989:70:205-212, 4 Stewart JR, Blackwell WH, Crute SL, Laughlin V, Hess ML. Greenfield LJ, Prevention of myocardial ischemia reperfusion injury with oxygen free radical scavangers. Surg. Forum 1982;33:317-320. 5 Meerson FZ, Kagan VE, Kozlov VP, Belkiss LM, Arkhipenko YV. The role of lipid peroxidation in pathogenesis of ischaemic damage and the antioxidant protection of the heart. Basic Res Cardioi 1982;77:465-485. 6 Rdth E, T&ok B, Kelemen D, Pollak 2s. Effect of antioxidant treatment during healing of experimental myocardial infarction. In: Matkovics B, Karmazsin L, Kalrisz H. Radicals, Ions and the Tissue Damage, Akademiai Kiado, Budapest 1990;213-220. 7 Loeper J, Groy J, Rozensztajn L, Bedu 0, Misson P. Lipid peroxidation and protective enzymes during myocardial infarction. Clin Chim Acta 1991;196:I 19-126. 8 Misra HP, Fridovic I. The role of superoxide anion in the autoxidant of epinephrine and a simple assay for superoxide of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 1972;247:3170-3175.
415 9 Matkovics B, Novak R, Hoang Due Hahn, Szabd L, Varga SzI, Zalesna G. A comparative study on some more important experimental animal peroxide metabolism enzymes. Comp Biochem Physiol 1977;568:31-34. 10 Beers RF, Jr, Sizer TW. Spectrophotometry for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 1952;195:133-140. 11 Matkovics B, Szabo L, Varga SzI. Determination of enzyme activities in lipid peroxidation and glutathione pathways. Laboratoriumi Diagnosztika 1988;15:248-250 (in Hungarian with English summary). 12 Little C, O’Brien PJ. An intracellular GSH-peroxidase with a lipid peroxide substrate. BBRC 1968;31:145-150. 13 Chiu DTY, Stults FH, Tappal AL. Purification and properties of rat lung soluble glutathione peroxidase. BBA 1976;445:558-566. 14 Sedlak I, Lindsay RH. Estimation of total, protein-bound and non-protein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem 1968;25:192-205. 15 Placer ZA, Cushman L, Johson BC. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical system. Anal Biochem -1966;16:359-364. 16 Lowry OH, Rosebrough NJ, Farr AL, Randall RN. Protein measurement with the Folin-phenol reagent. J Biol Chem 1951;193:265-275. 17 Gromadzinsky J, Sklodovska M. Erythrocyte glutathione peroxidase and myocardial infarction. JAMA 1990;263:949-950. 18 Kok JF, Hofman A, Wittman JCM. Decreased selenium levels in acute myocardial infarction JAMA 1989;261:1161-1164.
Correspondence Hungary.
to: A. Lbszlo, A. Szent-Gyorgyi, Medical University, Department of Pediatrics, Szeged,
413
CCA 05188
Letter
to the Editor
Changes in lipid peroxidation and antioxidant enzyme activity of human red blood cells after myocardial infarction (Received 5 December 1990, revision received 8 October 1991; accepted 15 October 1991)
Dear Editor, If lipid peroxides are increasingly produced in pathological processes the end effect might be thrombosis or cell necrosis [1,2]. Ischemic-reperfusion has been thoroughly studied as a potential source of oxygen radicals. The fundamental importance of oxygen radicals in this processes is also demonstrated in an indirect way by the effectioness of antioxidant therapy [3-71. We have further examined the mechanisms of reperfusion injury and now here to report how lipid peroxidation and antioxidant enzyme activity change 8 h after acute myocardial infarction. Activities of the antioxidant enzymes, superoxide dismutase (SOD; EC 1.15.1.1), catalase (Case; EC 1.11.1.6), glutathione peroxidase (GP-ase; EC 1.11.1.9) and the lipid peroxidation (LPI products were measured in haemolysates of red blood cells (RBCs) harvested from blood taken from the antecubital vein. The methods for these assays were: (i) SOD assay based on the inhibition of epinephrine-adrenochrome transformation [8,9]; (ii) catalase through the rate of H,O, cleavage/unit of time at 240 nm (9,101; (iii) activity of glutathione peroxidase with the help of cumen hydroperoxide co-substrate [ll-141; (iv) LP as total-thiobarbituric acid active material, and expressed as a malondialdehyde (MDA) value, using malondialdehyde diethylacetale for calibration 111,151; (v) protein content by the method of Lowry et al. [16]. Data were analysed for statistical significance with the Student’s t test. The values given are mean f SD. The results are given in Table I. Activities of all the antioxidant enzymes tested decreased. in RBC haemolysates from blood taken 8 h after acute myocardial infarct. While the changes in C-ase and SOD were significant the decrease in GP-ase was not. The LP value showed a pronounced increase [7]. The observed changes in GP-ase activity presented here in the initial period of cardiac infarct similar to those published by Gromadinska et al. 1171.It is intriguing that other observed the increase in GP-ase activity in RBCs after cardiac infarct [18]. It remains a question whether the above mentioned differences may come from the differences in the methods applied in measuring of GP-ase.
414 TABLE I RBC lipid peroxidation and antioxidant (mean f SD1 enzyme activities in myocardial infarction Erythrocyte haemolysate
LP nmol MDA/ml
Myocardial infarct SD *
101.3 2.5.6
2.30 0.61
< 0.01 67.4 15.9
> 0.05 2.57 0.26
group (n = 10) P Control group SD* (n=ll)
GP-ase U/ml haemoi.
SOD U/ml haemol.
Catalase BIJ/ml haemol
493.0 135.0
0.32
< 0.001 716.5 92.6
1.05
< 0.001 2.05 0.68
BU = Bergmeyer unit
Sjgnificant increase in RBC LP after myocardia~ infarct may also have a role in the inhibition of SOD and C-ase activities which results further H,O, accumulation and in a circulus viciosus which triggers again a decrease in enzyme activity. The H,O, accumulation shifting the arachidonic acid metabolism away from the prostacyclin parthway. Aranka L.&lo I A. Gent-Gy6rgyi
‘, B&a Matkovics 2, Sz. Ilona Varge 2, Tibor Wittman 3 and Tamas Fazekas 3 Me&al University, Department of Pediatrics, Szeged,
’ Biological Isotope Labortory, ‘Al ’ University of Szeged and s A. Szent-Gyiirgi Medical U~.~ersi~, 1st De~rt~eat of Internal Medicine, Szeged, Hungary
1 De Maestro RF. An approach to free radicals in medicine and biology. Acta Physiol Stand 1980; Suppl. 492:153-168. 2 McCord J, Fridovich I. The biology and pathology of oxygen radicals. Ann Intern Med 1978;89:122127. 3 Larner AJ, Conway MA. Free radicals in acute myocardial infarction. Quart J Med New Ser 1989:70:205-212, 4 Stewart JR, Blackwell WH, Crute SL, Laughlin V, Hess ML. Greenfield LJ, Prevention of myocardial ischemia reperfusion injury with oxygen free radical scavangers. Surg. Forum 1982;33:317-320. 5 Meerson FZ, Kagan VE, Kozlov VP, Belkiss LM, Arkhipenko YV. The role of lipid peroxidation in pathogenesis of ischaemic damage and the antioxidant protection of the heart. Basic Res Cardioi 1982;77:465-485. 6 Rdth E, T&ok B, Kelemen D, Pollak 2s. Effect of antioxidant treatment during healing of experimental myocardial infarction. In: Matkovics B, Karmazsin L, Kalrisz H. Radicals, Ions and the Tissue Damage, Akademiai Kiado, Budapest 1990;213-220. 7 Loeper J, Groy J, Rozensztajn L, Bedu 0, Misson P. Lipid peroxidation and protective enzymes during myocardial infarction. Clin Chim Acta 1991;196:I 19-126. 8 Misra HP, Fridovic I. The role of superoxide anion in the autoxidant of epinephrine and a simple assay for superoxide of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 1972;247:3170-3175.
415 9 Matkovics B, Novak R, Hoang Due Hahn, Szabd L, Varga SzI, Zalesna G. A comparative study on some more important experimental animal peroxide metabolism enzymes. Comp Biochem Physiol 1977;568:31-34. 10 Beers RF, Jr, Sizer TW. Spectrophotometry for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 1952;195:133-140. 11 Matkovics B, Szabo L, Varga SzI. Determination of enzyme activities in lipid peroxidation and glutathione pathways. Laboratoriumi Diagnosztika 1988;15:248-250 (in Hungarian with English summary). 12 Little C, O’Brien PJ. An intracellular GSH-peroxidase with a lipid peroxide substrate. BBRC 1968;31:145-150. 13 Chiu DTY, Stults FH, Tappal AL. Purification and properties of rat lung soluble glutathione peroxidase. BBA 1976;445:558-566. 14 Sedlak I, Lindsay RH. Estimation of total, protein-bound and non-protein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem 1968;25:192-205. 15 Placer ZA, Cushman L, Johson BC. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical system. Anal Biochem -1966;16:359-364. 16 Lowry OH, Rosebrough NJ, Farr AL, Randall RN. Protein measurement with the Folin-phenol reagent. J Biol Chem 1951;193:265-275. 17 Gromadzinsky J, Sklodovska M. Erythrocyte glutathione peroxidase and myocardial infarction. JAMA 1990;263:949-950. 18 Kok JF, Hofman A, Wittman JCM. Decreased selenium levels in acute myocardial infarction JAMA 1989;261:1161-1164.
Correspondence Hungary.
to: A. Lbszlo, A. Szent-Gyorgyi, Medical University, Department of Pediatrics, Szeged,
