RAT ALT EKSTREMİTE İSKEMİ-REPERFÜZYON MODELİNDE İSKEMİK ÖNKOŞULLAMA VE ARDKOŞULLAMANIN ERKEN DÖNEM ETKİLERİ

Year 2011, Volume: 12 Issue: 3, 21 - 27, 01.12.2011

Tuğrul Ünsal Güneş Muharrem İsmail Dabak Tünay Kurtoğlu Aslıhan Karul Erdem Ali Özkısacık Berent Dişcigil

Abstract

AMAÇ: Bu çalışmanın amacı kardiyovasküler cerrahide önemli bir morbidite ve mortalite nedeni olan iskemireperfüzyon (İR) hasarına karşı iskemik ön ve ard koşullama ile bunların birlikte kullanılmasının koruyucu etkilerinin rat alt ekstremitesinde oluşturulan deneysel modelde araştırılmasıdır. GEREÇ ve YÖNTEM: Erkek Wistar-Albino ratları 5 gruba (n=40) ayırıldı. Sham grubunda (n=8) laparotomi yapıldı ancak iskemi oluşturulmadı. Kontrol (İR) grubunda (n=8) infrarenal aort 120 dk. süreyle klemplenerek oklüde edilmek yoluyla iskemi ve ardından 60 dk. süreyle reperfüzyon oluşturuldu. İskemik ard koşullama grubunda (n=8) reperfüzyon döneminin başlangıcında 5 dk. süreyle iskemi ve takiben 5 dk. reperfüzyon oluşturuldu ve bu işlem üç kez tekrar edildikten sonra reperfüzyon, kontrol grubunda olduğu şekilde tamamlandı. İskemik ön koşullama grubunda (n=8) uzun süreli iskemiden önce üç kez tekrarlanacak şekilde 10 dk.'lık iskemiyi takip eden 10 dk. süreli reperfüzyon oluşturulduktan sonra İR meydana getirildi. İskemik ön koşullama ile birlikte iskemik ard koşullama grubunda (n=8) İR periyodu öncesinde üç siklus şeklinde 10 dk. iskemi ve 10 dk. reperfüzyon oluşturuldu ve reperfüzyon döneminin başlangıcında işlem üç siklus şeklinde 5 dk.iskemi ve 5 dk. reperfüzyon olarak tekrarlandı. Reperfüzyon periyodunun sonunda ratların sol gastroknemius adalelerinden malondialdehit (MDA) ve nitrik oksit (NO) ölçümleri için doku örneği ve serum sitokin (TNF-α, IL 1-β, IL-6) değerlerinin belirlenmesi için kan örneği alındı.BULGULAR: Çalışma gruplarında MDA düzeylerinin kontrol grubuna göre daha düşük düzeyde ve ard koşullama grubunda sonuçların istatistiksel olarak anlamlı olduğu görülmüştür. İstatistiksel olarak anlamlı olmasa da en düşük TNF-α düzeyi ard koşullama grubunda saptanmıştır. İnterlökin 1-β düzeylerinin tüm gruplarda kontrol grubuna göre anlamlı olarak daha düşük olduğu gözlemlenmiştir. Ön koşullama ve ön koşullama ile birlikte ard koşullama gruplarında MDA düzeyleri kontrol grubuna göre düşük olmasına rağmen anlamlı bulunmamıştır.SONUÇ: İskemik koşullama modellerinde kontrol grubuna göre istatistiksel olarak anlamlı olmamasına rağmen erken dönemde doku hasarının daha az olduğunu gözlemledik. İskemik koşullamanın etkilerinin aydınlatılması için daha ileri çalışmalara ihtiyaç olduğunu düşünmekteyiz

Keywords

İskemi-reperfüzyon, ön koşullama, ard koşullama, sitokin

Effects of Ischemic Preconditioning and Postconditioning in a Rat Model of Lower Extremity Ischemia-Reperfusion in Early Phase

Abstract

PURPOSE: Ischemia-reperfusion injury is a major cause of morbidity and mortality in cardiovascular surgery. The aim of this study was to evaluate the protective effects of ischemic pre- and post conditioning and concomittant application of these two methods in a rat model of lower extremity ischemia-reperfusion (IR). MATERIALS and METHODS: Male Wistar-Albino rats (n=40) were divided into five groups. Sham group (n=8) underwent laparotomy but no ischemia. The infrarenal aorta was cross-clamped for 120 min. followed by 60 min. reperfusion in control (IR) (n=8) group. In ischemic post conditioning group (n=8) a period of ischemia for 5 min. followed by reperfusion for 5 min.repeated three times was formed at the beggining of repefusion. Three cycles of 10 min. of ischemia followed by 10 min of reperfusion was formed before IR in the ischemic preconditioning (n=8) group. In an other group synchronous application of pre- and post conditioning (n=8) was achieved. At the end of the reperfusion period, biopsy of the left gastrocnemius muscle was performed for determination of tissue malondialdehyte (MDA) and nitric oxide (NO) levels and blood samples were withdrawn for analysing serum cytokine (TNF-α, IL 1-β, IL-6) levels. RESULTS: Tissue MDA levels were observed to be lower as compared with the control group and it was significantly reduced in the post-conditioning group. TNF-α level was lowest in the post-conditioning group. Interleukin 1-β was found to be significantly lower in all groups as compared with the control group. In the preconditioning and synchronous pre- and postconditioning groups MDA levels were lower than the level in control group. CONCLUSION: Although the results were not statistically significant it was observed that there was a tendency towards reduced tissue injury in models of ischemic conditioning. These findings suugested that further studies are needed in order to clarify the effects of ischemic conditioning.

Keywords

Ischemia-reperfusion, preconditioning

References

• Konukoglu D, Hatemi HH, Arikan S, Demir M, Akcay T. Radioiodine treatment and oxidative stress in thyroidectomised patients for differentiated thyroid cancers. Pharmacol Res 1998; 4:311-5.
• Cortas NK, Wakid NW. Determination of inorganic nitrate in serum and urine by a kinetic cadmium- reduction method. Clin Chem. 1990; 36:1440-3.
• Hausenloy DJ, Yellon DM. The therapeutic potential of ischemic conditioning: an update. Nat Rev Cardiol Jun 21. Doi:10.1038/nrcardio.2011.85
• Murry CE, Jennings RB, Reimer KA. Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. Circulation 1986;74:1124-36.
• Mousney RA, Pang CY, Boyd JB, Forrest C. Augmentation of skeletal muscle survival in the latissimus dorsi porcine model using acute ischemic preconditioning. J Otolaryngol 1992; 21:315-20.
• Pasupathy S, Homer-Vanniasinkham S. Surgical implications of ischemic preconditioning. Arch Surg ; 140:405-9. Saita Y, Yokoyama K, Itoman M. Protective effect of ischaemic preconditioning against ischaemia-induced reperfusion injury of skeletal muscle: how many preconditioning cycles are appropriate? Brit J Plast Surg 2002; 55:241-5.
• Badhwar A, Bihari A, Dungey AA. Protective mechanisms during ischemic tolerance in skeletal muscle. Free Rad Biol Med 2004; 36:371-9.
• Küntscher MV, Schrimbek EU, Menke H: Ischemic preconditioning by brief extremity ischemia before flap ischemia in a rat model. Plastic and Reconst Surg ;109: 2398-404.
• Kin H, Zhao Z-Q, Sun H-Y, et al. Postconditioning attenuates myocardial ischemia-reperfusion injury by inhibiting events in the early minutes of reperfusion. Cardiovasc Res 2004;62:74-85.
• Hausenloy DJ, Lecour S, Yellon DM. Reperfusion injury salvage kinase and survivor activating factor enhancement prosurvival signaling pathways in ischemic postconditioning: two sides of the same coin. Antioxid Redox Signal. 2011; 14:893-907.
• Luna-Ortiz P, Torrres JC, Pastelin G, Martínez-Rosas M. Myocardial postconditioning: anaesthetic considerations. Arch Cardiol Mex 2011;81:33-46.
• Knudsen AR, Kannerup AS, Dich R, Kruhİffer M, Funch-Jensen P, Grİnbaek H, Mortensen FV. Expression of genes involved in rat liver angiogenesis after ischaemia and reperfusion: effects of ischaemic pre- and post-conditioning. HPB (Oxford). 2010;12 :554-60.
• Zhang F, Oswald T, Holt J. Regulation of inducible nitric oxide synthase in ischemic preconditioning of muscle flap in a rat model. Ann Plast Surg ;52:609-13. Zhang WX, Yin W, Zhang L, Wang LH, Bao L, Tuo HF, Zhou LF, Wa ng CC. Preconditioning and postconditioning reduce hepatic ischamia-reperfusion injury in rats. Hepatobiliary Pancreat Dis Int 2009; :586-90. injury and cardioprotective mechanisms: Role of mitochondria and reactive oxygen species. World J Cardiol 2011; 3:186-200.
• Hasturk A, Atalay B, Calısaneller T, Ozdemir O, Oruckaptan H, Altınors N. Analaysis of serum pro- inflammatory cytokine levels after rat spinal cor ischemia-reperfusion injury and correlation with tisuue damage. Turkish Neurosurgery 2009;19:353-359.
• Kellog EW. Superoxide, hydrogen peroxide and singlet Garcia-Dorado D, Barba I, Inserte J. Twenty-five years of preconditioning: are we ready for ischemia? From coronary occlusion to systems biology and back. Cardiovasc Res 2011;91:378-81. oxygen in lipid peroxidation by a xanthine system. J Biol Chem 1975; 250:8812.
• Slater TF. Free radical mechanisms in tissue injury. Biochem J 1984; 222: 1.
• Ignarro LJ. Biological actions and properties of endothelium-derived nitric oxide formed and released from artery and vein. Circ Res 1989;65:1.
• Yuan Y, Granger HJ. Flow modulates coronary venular permeability by a nitric oxide-related mechanism. Am J Physiol 1992;263:H641.
• Kubes P, Granger DN. Nitric oxide modulates microvascular permeability. Am J Physiol ;262:H611. Johnson III G, Tsao PS, Lefer AM. Cardioprotective effects of authentic nitric oxide in myocardial ischemia with reperfusion. Crit Carre Med 1991;19:244-52.
• Lefer DJ, Nakanishi K, Johnston WE, et al. Anti- neutrophil and myocardial protecting action of SPM- , a novel nitric oxide (NO) donor, following acute myocardial ischemia and reperfusion in dogs. Circulation 1991;88:2337-50. YAZIŞMA ADRESİ
• Doç. Dr. Muharrem İsmail BADAK Adnan Menderes Üniversitesi Tıp Fakültesi, Kalp ve Damar Cerrahisi AD, AYDIN, TÜRKİYE Telefon E-Posta : [e-mail] Geliş Tarihi Kabul Tarihi : 22.09.2011