Quercetin'in Sıçanlarda Alt Ekstremite İskemi Reperfüzyonu Sonrası Akciğer Hasarı Üzerindeki Etkilerinin Değerlendirilmesi

Öz

Özet Amaç: Quercetin antioksidan ve antiinflamatuar etkilere sahiptir. Literatürde daha önce Quercetin'in iskemi reperfüzyon hasarı üzerindeki etkisini araştıran hayvan deneyleri olmasına rağmen, alt ekstremite IR'nin uzak organ üzerindeki etkisini içeren çalışmalar nadirdir. Bu nedenle alt ekstremite iskemi reperfüzyon hasarı olan sıçanlarda Quercetin'in uzak organ (akciğer) üzerindeki koruyucu etkilerini araştırdık. Materyal ve Metod: 18 adet erkek Wistar Albino rat, her grupta 6 adet olmak üzere rastgele 3 gruba ayrıldı; Kontrol (C), İskemi-reperfüzyon (IR), IR-Quercetin, (IR-Q). Ağırlıkları 200-250 gr arasındaydı. İşlemden 30 dakika önce intraperitoneal olarak 20 mg.kg-1 Quercetin verildi. iskemi reperfüzyon gruplarında infrarenal abdominal aorta atravmatik mikrovasküler klemp ile klemplendi. 120 dakika sonra klempi çıkardık ve 120 dakika daha reperfüzyon sağlandı. Reperfüzyon süresi sona erdiğinde akciğerlerden doku örnekleri alındı. Malondialdehit (MDA) düzeyi, süperoksit dismutaz (SOD) ve katalaz (CAT) enzim aktivitesi ve histopatolojik parametreler karşılaştırıldı. Bulgular: İR grubunda MDA düzeyini kontrol grubuna göre daha yüksek bulduk (p<0,0001). IR-Q grubunda IR grubuna göre daha düşük MDA düzeyi bulundu (p=0.012). IR grubundaki SOD ve CAT enzim aktivitesi kontrol grubunda belirgin şekilde daha düşüktü (sırasıyla p<0.0001, p<0.001). IR-Q grubunda IR grubuna göre daha yüksek SOD ve CAT enzim aktiviteleri bulundu (sırasıyla p=0.012, p<0.001). Nötrofil infiltrasyonu/agregasyonu, alveolar duvar kalınlığı ve toplam akciğer hasarı skoru, IR grubunda C grubuna göre belirgin olarak daha yüksekti (sırasıyla p=0,001, p=0,002, p<0,0001). Ayrıca, Quercetin ile tedavi edilen grupta nötrofil infiltrasyonu/agregasyonu, alveolar duvar kalınlığı ve toplam akciğer hasarı skorunda IR grubuna göre istatistiksel olarak anlamlı bir düşüş gözlendi (sırasıyla p=0.023, p=0.022, p=0.002). Sonuç: Ratlarda iskemiden 30 dakika önce 20 mg.kg-1 intraperitoneal olarak uygulanan Quercetin'in lipid peroksidasyonunu ve oksidatif stresi azalttığını ve akciğer histopatolojisinde IR'nin neden olduğu hasarı düzelttiğini belirledik. Quercetin'in IR öncesi uygulandığında akciğer koruyucu etkisi olduğunu düşünüyoruz. Çalışmanın bulguları diğer çalışmalarla desteklendiğinde Quercetin'in IR hasarı üzerindeki koruyucu etkilerinin detaylı olarak ortaya konacağını ve kullanım endikasyonlarının genişleyeceğini düşünüyoruz.

Anahtar Kelimeler

• Quercetin
• Reperfüzyon Hasarı
• Akciğer
• Malondialdehit
• Süperoksit Dismutaz
• Katalaz

Assessment of the Effects of Quercetin on Lung Injury After Hind Limb Ischemia Reperfusion in Rats

Abstract

Background: Quercetin has antioxidant and anti-inflammatory effects. Although there are previous animal experi-ments investigating Quercetin’s effect on ischemia reperfusion (IR) injury in the literature, studies involving effect of lower extremity IR on remote organ are rare.
Materials and Methods: 18 male Wistar Albino rats were randomly divided into 3 groups, 6 in each group as; Cont-rol (C), Ischemia-reperfusion (IR), IR-Quercetin, (IR-Q). Their weights were between 200-250 g. 30 minutes before the procedure20 mg/kgQuercetin was administered via intraperitoneal route. In the IR groups, infrarenal abdomi-nal aorta was clamped by an atraumatic microvascular clamp. After 120 minutes of ischemia and reperfusion was achieved for another 120 minutes. When reperfusion period ended, tissue samples were taken from the lungs. Malondialdehyde (MDA) level, superoxide dismutase (SOD) and catalase (CAT) enzyme activity and histopathological parameters were compared.
Results: We found the MDA level in the IR group higher than the control group (p<0.0001). Lower MDA level was found in the IR-Q group compared tothe IR group (p=0.012). SOD and CAT enzyme activity in the IR group was notably lower in the control group (p<0.0001, p<0.001, respectively). Higher SOD and CAT enzyme activities were found in the IR-Q group compared to the IR group (p=0.012, p<0.001, respectively). Neutrophil infiltra-tion/aggregation, alveolar wall thickness and total lung injury score were notably higher in IR group than in C group (p=0.001, p=0.002, p<0.0001, respectively). In addition, a statistically significant decrease was observed in the Quercetin treated group in neutrophil infiltration/aggregation, alveolar wall thickness and total lung injury score compared to the IR group (p=0.023, p=0.022, p=0.002, respectively).
Conclusions: We determined that intraperitoneally administered Quercetin at a dose of 20 mg/kg30 minutes before ischemia in rats reduces lipid peroxidation, oxidative stress and reduces the damage caused by IR in lung histo-pathology.Study findings suggest that Quercetin has a lung protective effect when administered before IR.

Keywords

• Quercetin
• Reperfusion Injury
• Lung
• Malondialdehyde
• Superoxide Dismutase
• Catalase

References

1. Referans1. Blaisdell FW. The pathophysiology of skeletal muscle ischemia and the reperfusion syndrome: a review. Cardiovasc Surg 2002;10:620–30.
2. Referans2. Earnshaw JJ. Demography and etiology of acute leg ischemia. Semin Vasc Surg 2001;14:86–92.
3. Referans3. Sotoudeh A, Takhtfooladi MA, Jahanshahi A, Asl AHK, Takhtfooladi HA, Khansari M. Effect of N-acetylcysteine on lung injury induced by skeletal muscle ischemia-reperfusion. Histopathlogical study in rat model. Acta Cir Bras 2012;27:168–71.
4. Referans4. Takhtfooladi MA, Jahanshahi A, Jahanshahi G, Sotoudeh A, Takhtfooladi HA, Khansari M. Protective effect of N-acetylcysteine on kidney as a remote organ after skeletal muscle ischemia-reperfusion. Acta Cir Bras. 2012 Sep;27(9):611-5. doi: 10.1590/s0102-86502012000900004. PMID: 22936085.
5. Referans5. Köksal E, Gülçin I, Beyza S, Sarikaya O, Bursal E. In vitro antioxidant activity of silymarin. J Enzyme Inhib Med Chem. 2009 Apr;24(2):395-405. doi: 10.1080/14756360802188081. PMID: 18830883.
6. Referans6. Eltzschig HK, Collard CD. Vascular ischaemia and reperfusion injury. Br Med Bull 2004;70:71-86.
7. Referans7. Pedro C, Susimeire G. Ischemia/reperfusion-induced lung injury prevention: many options, no choices. J Bras Pneumol. 2016;42(1):7-8.
8. Referans8. Takhtfooladi H, Takhtfooladi M, Moayer F, Mobarakeh S. Melatonin attenuates lung injury in a hind limb ischemia-reperfusion rat model. Rev Port Pneumol (2006). 2015 Jan-Feb;21(1):30-5. doi: 10.1016/j.rppnen.2014.01.010. Epub 2015 Jan 20. PMID: 25854133.

9. Referans9. Takhtfooladi MA, Takhtfooladi HA, Sedaghatfar H, Shabani S. Effect of low-level laser therapy on lung injury induced by hindlimb ischemia/reperfusion in rats. Lasers Med Sci. 2015 Aug;30(6):1757-62. doi: 10.1007/s10103-015-1786-6. Epub 2015 Jul 9. PMID: 26155904.
10. Referans10. Kalin P, Gulcin I, Goren AC. Antioxidant activity and polyphenol content of cranberries (Vaccinium macrocarpon). Rec Nat Prod 2015;9:496–502.
11. Referans11. Sehitoglu MH, Han H, Kalin P, Gülçin İ, Ozkan A, Aboul-Enein HY. Pistachio (Pistacia vera L.) gum: a potent inhibitor of reactive oxygen species. J Enzyme Inhib Med Chem. 2015 Apr;30(2):264-9. doi: 10.3109/14756366.2014.915395. Epub 2014 Jun 18. PMID: 24939094.
12. Referans12. Gulcin I. In vitro prooxidant effect of caffeine. J Enzym Inhib Med Chem 2008;23:149–52.
13. Referans13. Cetinkaya Y, Göçer H, Menzek A, Gülçin I. Synthesis and antioxidant properties of (3,4-dihydroxyphenyl)(2,3,4-trihydroxyphenyl)methanone and its derivatives. Arch Pharm (Weinheim). 2012 Apr;345(4):323-34. doi: 10.1002/ardp.201100272. Epub 2011 Dec 7. PMID: 22147548.
14. Referans14. Gülçin I, Elias R, Gepdiremen A, Chea A, Topal F. Antioxidant activity of bisbenzylisoquinoline alkaloids from Stephania rotunda: cepharanthine and fangchinoline. J Enzyme Inhib Med Chem. 2010 Feb;25(1):44-53. doi: 10.3109/14756360902932792. PMID: 20030508.
15. Referans15. Gülçin İ, Tel AZ, Kirecci E. Antioxidant, antimicrobial, antifungal and antiradical activities of Cyclotrichium niveum (Boiss.) Manden and Scheng. Int J Food Propert 2008;11:450–71.
16. Referans16. Sinha R, Srivastava S, Joshi A, Joshi UJ, Govil G. In-vitro anti-proliferative and anti-oxidant activity of galangin, fisetin and quercetin: role of localization and intermolecular interaction in model membrane. Eur J Med Chem. 2014 May 22;79:102-9. doi: 10.1016/j.ejmech.2014.04.002. Epub 2014 Apr 2. PMID: 24727463.
17. Referans17. Burak C, Wolffram S, Zur B, Langguth P, Fimmers R, Alteheld B, et al. Effect of alpha-linolenic acid in combination with the flavonol quercetin on markers of cardiovascular disease risk in healthy, non-obese adults: A randomized, double-blinded placebo-controlled crossover trial. Nutrition. 2019 Feb;58:47-56. doi: 10.1016/j.nut.2018.06.012. Epub 2018 Jul 12. PMID: 30278429.
18. Referans18. Chen YW, Chou HC, Lin ST, Chen YH, Chang YJ, Chen L, et al. Cardioprotective Effects of Quercetin in Cardiomyocyte under Ischemia/Reperfusion Injury. Evid Based Complement Alternat Med. 2013;2013:364519. doi: 10.1155/2013/364519. Epub 2013 Mar 14. PMID: 23573126; PMCID: PMC3612448.
19. Referans19. Gülçin İ, Elmastas M, Aboul-Enein HY. Determination of antioxidant and radical scavenging activity of basil (Ocimum basilicum) assayed by different methodologies. Phytother Res 2007;21:354–61.
20. Referans20. Kılıç Y, Özer A, Tatar T, Zor MH, Kirişçi M, Kartal H, et al. Effect of picroside II on hind limb ischemia reperfusion injury in rats. Drug Des Devel Ther. 2017 Jun 26;11:1917-1925. doi: 10.2147/DDDT.S132401. PMID: 28721011; PMCID: PMC5500556.
21. Referans21. Durak I, Canbolat O, Kavutçu M, Oztürk HS, Yurtarslani Z. Activities of total, cytoplasmic, and mitochondrial superoxide dismutase enzymes in sera and pleural fluids from patients with lung cancer. J Clin Lab Anal. 1996;10(1):17-20. doi: 10.1002/(SICI)1098-2825(1996)10:1<17::AID-JCLA4>3.0.CO;2-I. PMID: 8926562.
22. Referans22. Van Ye TM, Roza AM, Pieper GM, Henderson J Jr, Johnson CP, Adams MB. Inhibition of intestinal lipid peroxidation does not minimize morphologic damage. J Surg Res. 1993 Nov;55(5):553-8. doi: 10.1006/jsre.1993.1183. PMID: 8231175.
23. Referans23. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265-75. PMID: 14907713.
24. Referans24. Aebi H. Catalase. In: H.U. Bergmeyer (Ed): Methods of Enzymatic Analysis, Academic Press, New York and London, 1974; pp.673-677.
25. Referans25. Tuncay A, Sivgin V, Ozdemirkan A, Sezen SC, Boyunaga H, Kucuk A, et al. The Effect of Cerium Oxide on Lung Tissue in Lower Extremity Ischemia Reperfusion Injury in Sevoflurane Administered Rats. Int J Nanomedicine. 2020 Oct 6;15:7481-7489. doi: 10.2147/IJN.S263001. PMID: 33116483; PMCID: PMC7547779.
26. Referans26. Akbas H, Ozden M, Kanko M, Maral H, Bulbul S, Yavuz S, et al. Protective antioxidant effects of carvedilol in a rat model of ischaemia-reperfusion injury. J Int Med Res. 2005 Sep-Oct;33(5):528-36. doi: 10.1177/147323000503300508. PMID: 16222886.
27. Referans27. Abola MTB, Golledge J, Miyata T, Rha SW, Yan BP, Dy TC, et al. Asia-Pacific Consensus Statement on the Management of Peripheral Artery Disease: A Report from the Asian Pacific Society of Atherosclerosis and Vascular Disease Asia-Pacific Peripheral Artery Disease Consensus Statement Project Committee. J Atheroscler Thromb. 2020 Aug 1;27(8):809-907. doi: 10.5551/jat.53660. Epub 2020 Jul 4. Erratum in: J Atheroscler Thromb. 2020 Dec 1;27(12):1374. Erratum in: J Atheroscler Thromb. 2021;28(3):304. PMID: 32624554; PMCID: PMC7458790.
28. Referans28. Harkin DW, Barros D'sa AA, McCallion K, Hoper M, Halliday MI, Campbell FC. Circulating neutrophil priming and systemic inflammation in limb ischaemia-reperfusion injury. Int Angiol. 2001 Mar;20(1):78-89. PMID: 11455935.
29. Referans29. Zahedi M, Ghiasvand R, Feizi A, Asgari G, Darvish L. Does Quercetin Improve Cardiovascular Risk factors and Inflammatory Biomarkers in Women with Type 2 Diabetes: A Double-blind Randomized Controlled Clinical Trial. Int J Prev Med. 2013 Jul;4(7):777-85. PMID: 24049596; PMCID: PMC3775217.
30. Referans30. Birdane FM, Cemek M, Birdane YO, Gülçin I, Büyükokuroğlu ME. Beneficial effects of Foeniculum vulgare on ethanol-induced acute gastric mucosal injury in rats. World J Gastroenterol. 2007 Jan 28;13(4):607-11. doi: 10.3748/wjg.v13.i4.607. PMID: 17278229; PMCID: PMC4065985..
31. Referans31. Pérez JM, Arenas FA, Pradenas GA, Sandoval JM, Vásquez CC. Escherichia coli YqhD exhibits aldehyde reductase activity and protects from the harmful effect of lipid peroxidation-derived aldehydes. J Biol Chem. 2008 Mar 21;283(12):7346-53. doi: 10.1074/jbc.M708846200. Epub 2008 Jan 22. PMID: 18211903.
32. Referans32. Olguner C, Koca U, Kar A, Karci A, Işlekel H, Canyilmaz M, et al. Ischemic preconditioning attenuates the lipid peroxidation and remote lung injury in the rat model of unilateral lower limb ischemia reperfusion. Acta Anaesthesiol Scand. 2006 Feb;50(2):150-5. doi: 10.1111/j.1399-6576.2006.00938.x. PMID: 16430534.
33. Referans33. Draper HH, Hadley M. Malondialdehyde determination as index of lipid peroxidation. Methods Enzymol 1990;86:421–3.
34. Referans34. Henderson PW, Singh SP, Weinstein AL, Nagineni V, Rafii DC, Kadouch D, et al. Therapeutic metabolic inhibition: hydrogen sulfide significantly mitigates skeletal muscle ischemia reperfusion injury in vitro and in vivo. Plast Reconstr Surg. 2010 Dec;126(6):1890-1898. doi: 10.1097/PRS.0b013e3181f446bc. PMID: 21124129.
35. Referans35. Hazafa A, Rehman KU, Jahan N, Jabeen Z. The Role of Polyphenol (Flavonoids) Compounds in the Treatment of Cancer Cells. Nutr Cancer. 2020;72(3):386-397. doi: 10.1080/01635581.2019.1637006. Epub 2019 Jul 9. PMID: 31287738.
36. Referans36. Liu H, Guo X, Chu Y, Lu S. Heart protective effects and mechanism of quercetin preconditioning on anti-myocardial ischemia reperfusion (IR) injuries in rats. Gene. 2014 Jul 15;545(1):149-55. doi: 10.1016/j.gene.2014.04.043. Epub 2014 Apr 24. PMID: 24769323.
37. Referans37. Dok-Go H, Lee KH, Kim HJ, Lee EH, Lee J, Song YS, et al. Neuroprotective effects of antioxidative flavonoids, quercetin, (+)-dihydroquercetin and quercetin 3-methyl ether, isolated from Opuntia ficus-indica var. saboten. Brain Res. 2003 Mar 7;965(1-2):130-6. doi: 10.1016/s0006-8993(02)04150-1. PMID: 12591129.
38. Referans38. Zhang W, Wang Y, Yang Z, Qiu J, Ma J, Zhao Z, et al. Antioxidant treatment with quercetin ameliorates erectile dysfunction in streptozotocin-induced diabetic rats. J Biosci Bioeng. 2011 Sep;112(3):215-8. doi: 10.1016/j.jbiosc.2011.05.013. Epub 2011 Jun 14. PMID: 21664865.
39. Referans39. Sul OJ, Ra SW. Quercetin Prevents LPS-Induced Oxidative Stress and Inflammation by Modulating NOX2/ROS/NF-kB in Lung Epithelial Cells. Molecules. 2021 Nov 17;26(22):6949. doi: 10.3390/molecules26226949. PMID: 34834040; PMCID: PMC8625571.
40. Referans40. Huang R, Zhong T, Wu H. Quercetin protects against lipopolysaccharide-induced acute lung injury in rats through suppression of inflammation and oxidative stress. Arch Med Sci. 2015 Apr 25;11(2):427-32. doi: 10.5114/aoms.2015.50975. Epub 2015 Apr 23. PMID: 25995762; PMCID: PMC4424260.