Prevention of lung injury by iloprost following hind limb ischemia and reperfuison model in rats

Öz

Aim: The major part of tissue damage occurs upon reperfusion and is mediated by activated neutrophils that release oxygen free radicals. Following hind-limb ischemia/reperfusion, lung injury due to neutrophil infiltration and oxygen free radicals has been demonstrated. Previous studies have shown that this injury can be prevented pharmacologically. Iloprost is a long acting stable analog of prostacyclin. The aim of this study is to test the effect of iloprost in prevention of lung injury due to lower limb ischemia/reperfusion.

Material and Methods: Through a midline laparotomy infrarenal abdominal aorta was approached and cross-clamped in 20 male Spraque-Dawley rats for 2 hours. At the time of declamping Group I animals (n=8) received iloprost (0,1µg/kg/min) and Group II animals (n=8) received normal saline (0,1ml/kg/min) continously for 4 hours. Third group was the sham group (n=4). The lung tissue assays were performed for measurement of lipid peroxidation end product malondealdehyde and also total glutathione. Lung tissues were also examined histopathologically under light microscopy.

Results: The malondealdehyde levels in the iloprost group were significantly lower than the control group (p<0,05). The glutathione levels did not show any difference between the iloprost and the control groups (p>0,05). Histopathological examination revealed that the structure of the lung tissue was preserved in the iloprost group whereas lung tissue of the control group had evidence of injury.

Conclusion: Our results suggest that iloprost reduces the production of oxygen free radicals and prevents lung injury due to lower limb ischemia reperfusion.

Anahtar Kelimeler

• ischemia,reperfusion,iloprost

Sıçanlarda alt ekstremite iskemi reperfüzyon hasarını takiben gelişen akciğer hasarının iloprost ile önlenmesi

Öz

Amaç:  İskemi ve bunu takip eden reperfüzyon dönemi sonrasında ortaya çıkan doku hasarı

büyük ölçüde reperfüzyon sırasında gerçekleşir. Aktive olmuş nötrofillerden salınan serbest

oksijen radikalleri bu hasarda önemli rol oynarlar. Alt ekstremite iskemi-reperfüzyonu

sonrasında görülen akciğer hasarında nötrofil infiltrasyonunun ve serbest oksijen

radikallerinin önemi gösterilmiştir. Bu hasarı farmakolojik olarak önlemek için birçok çalışma

yapılmıştır. İloprost, uzun etkili bir prostasiklin analoğudur. Çalışmanın amacı, alt ekstremite

iskemi reperfüzyonu sonrası ortaya çıkan akciğer hasarını önlemedeki iloprostun etkisini

araştırmaktır.

Gereç ve Yöntemler: Çalışmada 20 adet Spraque-Dawley cinsi erkek sıçan kullanıldı. Orta hattan

yapılan laparotomi ile abdominal aortaya ulaşıldı. Grup I (n=8) ve Grup II (n=8) de bulunan

deneklerin infrarenal aortalarına 2 saat boyunca kross klemp uygulandı. Klemp kaldırıldığı

sırada Grup I’de bulunan deneklere 0,1 g/kg/dk dozunda iloprost ve Grup II’de bulunan

deneklere 0,1ml/kg/dk serum fizyolojik sürekli infüzyon şeklinde 4 saat boyunca verildi. Grup

III(n=4) sham grubu olarak belirlendi. Denekler 4. saatin sonunda sakrifiye edilerek akciğer

dokuları çıkarıldıve biyokimyasal ve histopatolojik inceleme yapıldı.

Bulgular: İloprost verilen grupta malondealdehyde seviyeleri kontrol grubuna göre

istatistiksel olarak anlamlı derecede düşük bulundu (p<0.05). Glutatyon düzeylerinde ise

kontrol ve iloprost grupları arasında anlamlı fark bulunmadı (p>0.05). Histopatolojik

incelemede, iloprost verilen grupta akciğer dokusunun korunmuş olduğu gözlenirken kontrol

grubunda akciğer doku harabiyeti tespit edilmiştir.

Sonuç: Sonuçlarımız alt ekstremite iskemi-reperfüzyonu sonrasında ortaya çıkan akciğer

hasarının önlenmesinde iloprostun serbest oksijen radikal oluşumunu azaltarak etkili

olduğunu göstermektedir.

Anahtar Kelimeler

• iloprost,iskemi,reperfüzyon,iloprost

Kaynakça

1. 1. Klausner JM, Paterson IS, Valeri R, Hechtman HB. Limb ischemia induced increase in permeability is mediated by leukocytes and leukotrienes. Ann Surg 1988; 208: 755-60.
2. 2. Klausner JM, Paterson IS, Kozbik L, Valeri R, Shepro D, Hetchtman HB. Oxygen free radicals mediate ischemia-induced lung injury. Surgery 1989; 105: 192-99.
3. 3. Gyurkovics E, Aranyi P, Stagnl R et al. Postconditioning of the lower limb protection against reperfusion syndrome. J Surg Res 2011; 169: 139- 47.
4. 4. Akgun S, Tekeli A, Isbir SC et al. FK506 to prevent lung injury after hindlimb ischemia and reperfusion in a rat model: an electron microscopic study. Surg Today 2004; 34: 678-84.
5. 5. Groeneveld AB, Raijmakers PG, Rauwerda JA, Hack CE. The inflammatory response to vascular surgery associated ischemia and reperfusion in man:effect of postoperative pulmonary function. Eur J Endovasc Surg 1997; 14: 351-59.
6. 6. Gaines GC, Welborn MB, Moldawer LL, Huber TS, Harward TR, Seeger JM. Attenuation of skeletal muscle ischemia-reperfusion by tumor necrosis factor. J Vasc Surg 1999; 29: 370-76.
7. 7. Bengisun U, Köksoy C, Bengisun JS, Bayraktaroglu G, Camur A, Aras N. Ischemia and reperfusion of pulmonary hypertension and leukosequestration following lower limb ischemia. Prost Leuko Essen Fatty Acids 1997; 56: 117-20.
8. 8. İsbir CS, Akgün S, Ak K et al. Akut alt ekstremite iskemi/reperfüzyon hasarının akciğer serbest oksijen radikalleri üzerine olan etkisi. TGKDCD 2000; 8: 629-31.

9. 9. Rocker MG. Ischemia/reperfusion, inflammatory responses and acute lung injury. Thorax 1997; 52: 841-42.
10. 10. Raijmakers PGHM, Groeneveld ABJ, Rauwerda JA, Teule GJJ, Hack CE. Acute lung injury after aortic surger: the relation between lung and leg microvascular permeability to indium-labelled transferrin and circulating mediators. Thorax 1997; 52: 866-71.
11. 11. Grant SM, Goa KL. Iloprost. A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic potential in peripheral vascular disease, myocardial ischaemia and extracorporeal circulation procedures. Drugs 1992; 43: 889-924.
12. 12. Wang L, Jin YZ, Zhao QH et al. Hemodynamic and gas exchange effects of inhaled iloprost in patients with COPD and pulmonary hypertension. Int J Chron Obstruct Pulmon Dis. 2017; 12: 3353-60.
13. 13. Pantazi E, Bejaoui M, Folch-Puy E, Adam R, Roselló-Catafau J. Advances in treatment strategies for ischemia reperfusion injury. Expert Opin Pharmacother. 2016; 17: 169-79.
14. 14. Bhattacharyya, A., Chattopadhyay, R., Mitra, S., Crowe, S.E., Oxidative stress: an essential factor in the pathogenesis of gastrointestinal mucosal diseases. Physiol. Rev. 2014; 94: 329–54.
15. 15. Messent M, Griffiths MJD, Evans TW. Pulmonary vascular reactivity and ischemia reperfusion injury in the rat. Clin Sci 1993; 85: 71–75.
16. 16. M. Sacar, V. Ozcan, H. Aybek et al., Vitamin C and iloprost attenuate skeletal muscle injury caused by ischemia-reperfusion of the lower extremities. TGKDCD 2005;13: 374–8.
17. 17. Aytacoglu BN, Sucu N, Tamer L et al., Iloprost for the attenuation of ischaemia/reperfusion injury in a distant organ. Cell Biochem Func 2006; 24: 341–46.
18. 18. Tiryakioglu O, Erkoc K, Tunerir B, Uysal O, Altin HF, Gunes T, Aydin S. The effect of iloprost and N-acetylcysteine on skeletal muscle injury in an acute aortic ischemia-reperfusion model: an experimental study. Biomed Res Int. 2015; 2015: 453748.