HİPOKSİ İLE OLUŞTURULAN MYOKARDİYAL HASAR ÜZERİNE MELATONİNİN ETKİSİ

Abstract

Sunulan çalışmada hipoksik hasarın etyopatogenezinde yer alan serbest radikallerin, lipid peroksidasyon ürünlerinin ve antioksidan enzimlerin rolünü değerlendirmek amaçlanmıştır. Çalışmada deney hayvanı olarak tavşan kullanıldı. Tavşanlar; Grup I: Hipoksi grubu (n=9), Grup II: Hipoksi+Melatonin grubu (n=9), Grup III: Kontrol grubu (n=7) olmak üzere üç gruba ayrıldılar. Hipoksi oluşturulmadan önce bazal serum troponin I, kreatin kinaz–MB (CKMB), laktat dehidrojenaz (LDH), plazma glutatyon peroksidaz (GSH-Px), süperoksit dismutaz (SOD), malondialdehit (MDA) ve nitrik oksit (NO) düzeyleri için kan alındı. Grup I, II’ deki deney hayvanları bir fanus içerisinde 5 lt/10 dk akımla %10 oksijen-azot karışımına maruz bırakılarak hipoksi oluşturuldu. İkinci grupta hipoksi oluşturulduktan sonra 5 gün süreyle melatonin (10 mg/kg/gün intraperitoneal) verildi. Tedavi sonunda serum CKMB, LDH ve troponin I, plazma NO, MDA, SOD ve GSH-Px düzeyleri çalışıldı. Tavşanlar sakrifiye edilip myokardiyal NO, MDA, SOD, GSH-Px düzeyleri çalışıldı ve histopatolojik olarak myokard dokusu Hemotoksilen-Eozin boyama ile değerlendirildi. Hipoksi oluşturulan birinci grup, tavşanlarda serum CKMB, LDH, troponin I düzeylerinde artış, plazma SOD’de, GSH-Px’da azalma ve histopatolojik olarak miyokard liflerinde şişme, intertisiyel ödem, disorganizasyon ve nekrozdan oluşan ağır kardiyomyopati tesbit edildi. Hipoksi+ Melatonin uygulanan grupta; plazma SOD düzeylerinde anlamlı artış, doku NO ve MDA düzeylerinde anlamlı düşme tesbit edildi. Histopatolojik incelemede, melatonin grubunda; miyokard liflerinde şişme, intertisiyel ödem ve disorganizasyondan oluşan hafif-orta kardiyomyopati tesbit edildi. Sonuç olarak; melatoninin antioksidan etkinlik sağlayarak hipoksiye bağlı kardiyak hasarı önleyebileceği kanısına varıldı

Keywords

• Hipoksi, myokardiyal hasar, malondialdehit, melatonin

Effects of Melatonin on the Myocardial Damage, Induced by Hypoxia

Abstract

The aim of this study was to evaluate the role of antioxidant enyzmes, lipid peroxidation products and free radicals which involve in the etiopathogenesis of the hypoxic damage. The experimental animals were assigned to 5 groups. Group 1: Hypoxia group (n=9), Group II: Hypoxia + melatonin group (n=9), Group III: Control group (n=7). Blood was taken for analysing the levels of basal serum troponin I, creatine kinase-MB (CKMB), lactate dehydrogenase(LDH), plasma glutation peroxidase (GSH-Px), superoxide dismutase (SOD), malondialdehyde (MDA) and nitric oxide (NO) before inducing the hypoxia. In the experimental animals of groups I, II, the hypoxia was induced by leaving the animals to breathe in belljar, containing 10 per cent oxygen-nitrogen mixture, for 10 minutes at 5 liter velocity. After inducing the hypoxia melatonin (10 mg/kg/daily & intraperitoneal) was administered to the animals of second group during 5 days. The levels of serum CKMB, LDH and troponin I, plasma NO, MDA, SOD and GSHPx were analysed at the end of the treatment. After sacrificing the experimental animals, tle levels of myocardial NO, MDA, SOD, GSH-Px were analysed and myocardial tissue was examined histopathologically with haematoxylin - eosin stain. The levels of serum-CMKB, LDH and troponin I were found increased; whereas the levels of plasma SOD and GSH-Px were found decreased in the hypoxia-induced group and a heavy cardiomyopathy associated with swelling in myocardial flaments, interstitial edema, disorganization and necrosis was determined. There was a meaningful increase in the levels of plasma SOD but a meaningful decrease in the levels of tissue NO and MDA and a mild and intermediate cardiomyopathy associated with swelling, interstitial edema and disorganization was shown histopathologically in the second group (hypoxia and melatonin). It is shown that the melatonin can protect the hypoxic cardiac damage by being activated as an antioxidant

Keywords

• Hypoxia, myocardial damage, malondialdehyde, melatonin

References

1. 1. Haider KH, Stimson WH. Cardiac myofibrillar proteins: biochemical markers to estimate myocardial injury. Mol Cell Biochem 1999,194:31-39.
2. 2. Dhalla NS, Elmoselhi AB, Hata T, Makino N. Status of myocardial antioxidants in ischemia-reperfusion injury. Cardivasc Res.2000,47:446-456.
3. 3. Das UN. Free radicals , cytokines and nitric oxide in cardiac failure and myocardial infarction. Mol Cell Biochem 2000,215:145- 152.
4. 4. Halliwell B. Free radicals and antioxidants: a personal view.Nutr Rev. 1994;52:253-265.
5. 5. Reiter RJ, Tan DX, Osuna C, Gitto E. Actions of melatonin in the reduction of oxidative stress. A review. J Biomed Sci 2000,7:444- 458.
6. 6. Reiter RJ, Carnerio RC. Melatonin in relation to cellular antioxidative defense mechanisms. Horm Metab Res 1997,29:363-372.
7. 7. Duncker DJ, Verdouw PD. Has melatonin a future as a cardioprotective agent? Cardiovasc Drugs Ther 2001,15:205-207.
8. 8. Rumsey WL, Abbott B, Bertelsen D et al. Adaptation to hypoxia alters enegy metabolism in rat heart. Am J Physiol 1999,276:71-80.

9. 9. Paglia DE, Valentina WN. Studies on the quantitavive and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 1967,70:158-169.
10. 10. Sun Y, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clin Chem 1988,3413:497-500.
11. 11. Jain SK. Evidence for membrane lipid peroxidation during the in vivo aging of human erytrocytes. Biochem Biophys Acta 1988,937:205-210.
12. 12. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissue by thiobarbutiric acid relations. Analytical Biochemistry 1979, 95:351-358.
13. 13. Davidge ST, Stranko CP, Roberts JM. Urine but not plasma nitric oxide metabolites are decreased in women with preeclampsia. Am J Obstet Gynecol 1996,174:1008-1013.
14. 14. Moshage H, Kok B, Huizenga JR, Jansen PL. Nitrite and nitrate determinations in plasma: a critical evaluation.Clin Chem 1995,41:892-896.
15. 15. Morishima I, Matsui H, Mukawa H, Hayashi K, Toki Y, Okumura K, Ito T, Hayakawa T. Melatonin, a pineal hormone with antioxidant property, protects against adriamycin cardiomyopathy in rats. Life Sci 1998,63:511-521.
16. 16. Foex P. Myocardial ischaemia. Clin Anaesth 1999,13:321-334.
17. 17. Jaffe AS, Landt Y, Parvin CA, et al. Comparative sensitivity of cardiac troponin I and lactate dehydrogenase isoenzymes for diagnosing acute myocardial infarction. Clinical Chemistry 1996, 42:1770-1776.
18. 18. Chiu A, Chan WK, Cheng SH, Leung CK, Choi CH. Troponin-I, myoglobin, and mass concentration of creatine kinase-MB in acute myocardial infarction. J Med 1999,92: 711- 718.
19. 19. Vorderwinkler KP, Mair J, Puschendorf B, Hempel A, Schlüter K.D, Piper H.M. Cardiac troponin I increases in parallel to cardiac troponin T ,creatine kinase and lactate dehydrogenase in effluents from isolated perfused rat hearts after hypoxiareoxygenation-induced myocardial injury. Clinica Chimica Acta 1996,251:113-117.X
20. 20. Remppis A, Scheffold T, Greten J et al. Intracellular compartmentation of troponin T: release kinetics after global ischemia and calcium paradox in the isolated perfused rat heart. J Mol Cell Cardiol 1995,27: 793-803.
21. 21. Asayama J, Yamahara Y, Otha B, et al. Release kinetics of cardiac troponin T in coronary effluent from isolated rat hearts during hypoxia and reoxygenation. Basic Res Cardiol 1992,87:428-436.
22. 22. Bleier J, Vorderwinkler KP, Falkensammer J e t a l . D i f f e r e n t i n t r a c e l l u l a r compartmentations of cardiac troponins and myosin heavy chains: a causal connection to their different early release after myocardial damage. Clin Chem 1998,44(9):1912-1918.
23. 23. Ferrari R, Ceconi C, Curello S, et al. Role of oxygen free radicals in ischemic and reperfused myocardium. J Clin Nutr 1991,53: 215-222.
24. 24. Rao PS, Cohen MV, Mueller HS. Production of free radicals and lipid peroxides in early experimental myocardial ischemia. J Mol Cell Cardiol 1983,15:713-716.
25. 25. Depre C, Fierain L, Hue L. Activation of nitric oxide synthase by ischaemia in the perfused heart. Cardiovasc Res 1997,33: 82– 87.
26. 26. Wang PH, Zweier JL. Measurement of nitric oxide and peroxynitrite generation in the postischemic heart: evidence for peroxynitrite-mediated reperfusion injury. J Biol Chem 1996,2 71: 29223–29230.
27. 27. Guarnieri C, Flamigni F, Caldarera CM. Role of oxygen in the cellular damage induced by re-oxygenation of hypoxic heart.J Mol Cell Cardiol 1980 ,12: 797-808.
28. 28. Kihlström M, Kainulainen H, Salminen A. Enzymatic and nonenzymatic lipid peroxidation capacities and antioxidants in hypoxic and reoxygenated rat myocardium. Exp Mol Pathol 1989;50:230-238.
29. 29. Arduini A, Mezzetti A, Porreca E et al. Effect of ischemia and reperfusion on antioxidant enzymes and mitochondrial inner membrane proteins in perfused rat heart. Biochim Biophys Acta 1988,970:113-121.
30. 30. Yuan SM. Cytochemistry and ultrastructure of canine myocardium undergoing global ischemia and reperfusion injury. J Med Sci 1999,15(1):1-7.
31. 31. Reiter R, Tang L, Garcia JJ, Munoz-Hoyos A. Pharmacological actions of melatonin in oxygen radical pathophysıology. Life Sci 1997,60:2255-2271
32. 32. Kaneko S, Okumuro K, Numaguchi Y, et al. Melatonin scavenges hydroxyl radical and protects isolated rat heart from ischemic reperfusion injury. Life Sci 2000,67:101- 112.
33. 33. Gilad E, Cuzzocrea S, Zingarelli B, Salzman AL, Szabo C. Melatonin is a scavenger of peroxynitrite. Life Sci 1997,60:169-174.
34. 34. Szarszoi O, Asemu G, Vanecek J, Ostadal B, Kolar F.Effects of melatonin on ischemia and reperfusion injury of the rat heart. Cardiovasc Drugs Ther 2001,15: 251-257.
35. 35. Sahna E, Acet A, Ozer MK, Olmez E. Myocardial ischemia-reperfusion in rats: reduction of infarct size by either sup ple me nta l p hys i olo gic al or pharmacological doses of melatonin. J Pineal Res 2002,33: 234-238.