N-acetyl cysteine attenuates ferroptosis mediated lung injury induced by lower limb ischaemia/reperfusion

Öz

Aim: This study aimed to analyze the effect of N-acetyl cysteine pretreatment on the prevention of ferroptosis mediated lung injury induced by lower limb ischemia and reperfusion. Material and Methods: Eighteen male Sprague-Dawley rats weighing 350-400 g were randomized into three groups. The animals received N-acetyl cysteine 150 mg/kg or normal saline 0.1 ml/kg intraperitoneally before the ischemic period. In the control and study groups, I/R injury was induced by clamping the aorta infrarenal for 2 hours, followed by 4 hours of reperfusion. The third group underwent sham surgery. After sacrifice, the lungs of the animals were extracted for both histopathological and biochemical analysis. Results: There was a significant difference between the control and study animals regarding tissue malondialdehyde (MDA), and glutathione (GSH) levels. In the control group, the MDA levels were increased and the GSH levels were increased significantly compared to the sham group that revealed a ferroptosis mediated lung injury. However, N-acetyl cysteine decreased the levels of MDA and increased the levels of GSH revealing a protective effect. The Prussian blue (free iron stain) staining which was used to examine iron deposition revealed a reduced deposition of iron in the N-acetyl cysteine group. Conclusion: The results of the present study suggest a protective effect of N-acetyl cysteine on ferroptosis mediated lung injury induced by lower limb ischemia-reperfusion in a rat model.

Anahtar Kelimeler

• ferroptosis
• N-ACETYL CYSTEINE
• ischemia/reperfusion

N-asetil sistein alt ekstremite iskemi/ reperfüzyonu tarafından indüklenen feroptosise bağlı akciğer hasarını azaltır

Öz

Amaç: Bu çalışma, N-asetil sistein ön tedavisinin, alt ekstremite iskemisi ve reperfüzyonun neden olduğu ferroptoza bağlı akciğer hasarının önlenmesi üzerindeki etkisini analiz etmeyi amaçlamıştır. Gereç ve Yöntemler: 350-400 g ağırlığında on sekiz erkek Sprague-Dawley sıçanı üç gruba randomize edildi. İskemik dönemden önce intraperitonal olarak 150 mg / kg N-Asetil Sistein veya normal salin 0.1 ml / kg verildi. Kontrol ve çalışma gruplarında, aort infrarenal düzeyde klemlenerek 2 saat boyunca I/R ve sonrasında 4 saat reperfüzyon indüklendi. Üçüncü gruba sham grubu olarak kullanıldı. Hayvanlarının hayatı sonlandırıldıktan sonra, histopatolojik ve biyokimyasal analiz için akciğerleri çıkarıldı. Bulgular: Doku Malondialdehit (MDA) ve glutatyon (GSH) seviyeleri arasında istatistiksel olarak anlamlı fark tespit edildi. Kontrol grubunda, MDA düzeyleri ve GSH düzeyleri, ferroptoza bağlı akciğer hasarı gösteren sham grubuna kıyasla önemli ölçüde arttı. Bununla birlikte, NAC alan grupta MDA seviyesi düşerken koruyucu bir etki olarak GSH seviyeleri yükseldi. Demir birikimini göstermek için kullanılan Prusya mavisi (serbest demir lekesi) boyaması, N-asetil sistein grubunda demir birikiminin azaldığını ortaya çıkarmıştır. Sonuç: Bu çalışmanın sonuçunda, bir sıçan modelinde alt ekstremite iskemisi-reperfüzyonunun neden olduğu ferroptozise bağlı akciğer hasarı üzerine N-asetil sisteinin koruyucu bir etkisi olduğunu düşündürmektedir.

Anahtar Kelimeler

• Feroptozis
• İskemi/ reperfüzyon
• n-asetil sistein

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: 192-9.
3. 3. Dixon SJ, Lemberg KM, Lamprecht MR et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell 2012; 149: 1060–72.
4. 4. Cao JY, Dixon SJ. Mechanisms of ferroptosis. Cell Mol Life Sci 2016; 73: 2195–209.
5. 5. Basit F, van Oppen LM, Schöckel L et al. Mitochondrial complex I inhibition triggers a mitophagy-dependent ROS increase leading to necroptosis and ferroptosis in melanoma cells. Cell Death Dis 2017; 8: 2716.
6. 6. Xie Y, Hou W, Song X et al. Ferroptosis: process and function. Cell Death Differ 2016; 23: 369–79.
7. 7. Angeli JPF, Shah R, Pratt DA, Conrad M. Ferroptosis inhibition: mechanisms and opportunities. Trends Pharmacol Sci 2017; 38: 489–98.
8. 8. Masaldan S, Bush AI, Devos D, Rolland AS, Moreau C. Striking while the iron is hot: Iron metabolism and ferroptosis in neurodegeneration. Free Radic Biol Med 2019; 133: 221–33.

9. 9. Liu P, Feng Y, Li H, Chen X, Wang G, Xu S, Li Y, Zhao L. Ferrostatin-1 alleviates lipopolysaccharide-induced acute lung injury via inhibiting ferroptosis. Cell Mol Biol Lett 2020; 25: 10.
10. 10. Li Y, Cao Y, Xiao J, Shang J, Tan Q, Ping F, Huang W, Wu F, Zhang H, Zhang X. Inhibitor of apoptosis-stimulating protein of p53 inhibits ferroptosis and alleviates intestinal ischemia/reperfusion-induced acute lung injury. Cell Death Differ 2020; 18: 2635-50.
11. 11. Vale JA, Meredith TJ, Crome P, Helliwell M, Volans GN, Widdop B, Goulding R. Intravenous N-acetylcysteine: the treatment of choice in paracetamol poisoning? Br Med J 1979; 2: 1435-6.
12. 12. Rendell R, Fairhall S, Graham S, Rutter S, Auton P, Smith A, Perrott R, Jugg B. Assessment of N-acetylcysteine as a therapy for phosgene-induced acute lung injury. Toxicol Lett 2018; 290: 145-52.
13. 13. Kalogeris T, Baines CP, Krenz M, Korthuis RJ. Cell biology of ischemia/ reperfusion injury. Int Rev Cell Mol Biol 2012; 298: 229–317.
14. 14. Ellman GL. Tissue sulphydryl groups. Arch Biochem Biophys 1959; 82: 70–7.
15. 15. Akgun S, Tekeli A, Isbir SC, Civelek A, Ak K, Sirvanci S, Arbak S, Yaylim I, Arsan S. FK506 to prevent lung injury after hindlimb ischemia and reperfusion in a rat model: an electron microscopic study. Surg Today 2004; 34: 678–84.
16. 16. Liu H, Drew P, Cheng Y, Visner GA. Pirfenidone inhibits inflammatory responses and ameliorates allograft injury in a rat lung transplant model. J Thorac Cardiovasc Surg 2005; 130: 852-8.
17. 17. Zimmerman BJ and Granger DN. Reperfusion injury. Surg Clin North Am 1992; 72: 65-83.
18. 18. Katseni K, Chalkias A, Kotsis T et al. The effect of perioperative ischemia and reperfusion on multiorgan dysfunction following abdominal aortic aneurysm repair. Biomed Res Int 2015; 2015: 598980.
19. 19. Kalogeris T, Bao Y, Korthuis RJ. Mitochondrial reactive oxygen species: a double edged sword in ischemia/reperfusion vs preconditioning. Redox Biol 2014; 2: 702-14.
20. 20. Imai H, Matsuoka M, Kumagai T, Sakamoto T, and Koumura T, “Lipid peroxidation-dependent cell death regulated by GPx4 and ferroptosis,” in Apoptotic and Nonapoptotic Cell Death, S. Nagata and H. Nakano, Eds., vol. 403 of Current Topics in Microbiology and Immunology, pp. 143– 170, Springer, Cham, 2017.
21. 21. Karuppagounder SS, Alin L, Chen Y et al. N-Acetylcysteine targets 5 Lipoxygenase- Derived, Toxic Lipids and Can Synergize With Prostaglandin E2 to inhibit Ferroptosis and Improve Outcomes Following Hemorrhagic Stroke in Mice. Ann Neurol 2018; 84: 854–872.
22. 22. Kao SJ, Wang D, Lin HI, Chen HI. n-Acetylcysteine abrogates acute lung injury induced by endotoxin. Clin Exp Pharmacol 2006; 33: 33–40.
23. 23. Guo DW, Wang CY, Shih HC. N-acetylcysteine and atorvastatin alleviates lung injury due to ischemia-reperfusion injury in rats. J Chin Med Assoc 2019; 82: 909-14.
24. 24. Kalimeris K, Briassoulis P, Ntzouvani A et al. N-acetylcysteine ameliorates liver injury in a rat model of intestinal ischemia reperfusion. J Surg Res 2016; 206: 263–72.