MATEM TUNÇDEMİR, Eda BÜYÜKÇOLPAN MİRZATAŞ, HAFİZE UZUN

Deneysel diyabetik nefropatide quercetin'in renoprotektif potansiyeli: antiapoptotik ve antioksidan etkilerin değerlendirilmesi

Amaç: Sıçanlarda diyabetin neden olduğu böbrek hasarına katkıda bulunan apoptoz düzenleyici genlerin ekspresyonunu değerlendirerek güçlü bir biyoflavonoid olan quercetin'in renoprotektif ve antiapoptotik etkilerini araştırmak.Yöntemler: Sıçanlar 4 gruba ayrıldı; Kontrol, STZ-diyabetik, STZ-diyabetik + Quercetin ve Quercetin kontrol. Açlık kan şekeri, vücut ağırlığı ve idrar mikroalbüminüri değerleri ölçüldü. İmmünohistokimya bax, bcl-2, kaspaz-3 antikorları kullanılarak gerçekleştirildi. Apoptoz tespiti için TUNEL yöntemi uygulandı. Homojenize böbrek dokularında ileri oksidasyon protein ürünleri (AOPP), protein karbonil oksidasyon (PCO), malondialdehid (MDA) ve süperoksit dismutaz (SOD) aktivitesi ölçüldü.Bulgular: Tedavi edilmeyen diyabetik gruba kıyasla, quercetin uygulanan diyabetik grupta kan şekeri ve mikroalbüminüri düzeyleri anlamlı olarak azalmıştı (sırasıyla; p=0,020, p=0,003). MDA, AOPP ve PCO seviyeleri anlamlı olarak azaldı (sırasıyla; p=0,001, p=0,0001, p=0,0005), ancak SOD aktivitesinin quercetin uygulanan diyabetik grupta arttığı tespit edildi (p=0,005). Quercetin uygulanan diyabetiklerde, tedavi edilmemiş diyabetik gruba kıyasla bcl-2, bax ve kaspaz-3'ün immün boyanması azaldı. Tedavi edilmeyen diyabetik grubun böbrek tübüllerinde apoptotik hücrelerde belirgin bir artış gözlenirken, quercetin uygulanan diyabetik grupta belirgin bir düşüş gözlendi (p=0,0001). Sonuç: Antioksidan etkileri olan quercetin'in antiapoptotik etkilerinin, diyabetik komplikasyonların etkilerini azaltmada ve yeni komplikasyonları önlemede yararlı olabileceği sonucuna vardık.

Anahtar Kelimeler:

Deneysel diyabetik nefropati, Quercetin, böbrek, apoptoz, oksidatif stres

Renoprotective potential of quercetin in experimental diabetic nephropathy: assesing antiapoptotic and antioxidant effects

Aim: We investigated renoprotective and anti-apoptotic effects of quercetin, a potent bioflavonoid, by evaluating expression of apoptosis-regulatory genes that contribute to the kidney damage caused by diabetes in rats. Methods: Rats were divided into 4 groups; Control, STZ-induced diabetic, STZ-induced diabetic+Quercetin and Quercetin control. Values of fasting blood glucose, body weight and urine microalbuminuria measured. Immunohistochemistry was performed using bax, bcl-2, caspase-3 antibodies. For apoptosis detection, TUNEL method was applied. Advanced oxidation protein products (AOPP), protein carbonyl oxidation (PCO), malondialdehyde (MDA) and superoxide dismutase (SOD) activity were measured in homogenized kidney tissues. Results: Blood glucose and microalbuminuria levels were significantly decreased in quercetin-treated diabetic group compared to the untreated-diabetic group (p=0.020 and p=0.003; respectively). MDA, AOPP and PCO levels were significantly decreased (p=0.001, p=0.0001 and p=0.0005; respectively); however, SOD activity were found to increase in quercetin-treated diabetic group (p=0.005). Immunostaining of bcl-2, bax and caspase-3 was decreased compared to the untreated-diabetic group. Apoptotic cells especially increased in the kidney tubuli of untreated-diabetic group and on the contrary, a significant decrease was observed in the group that received a quercetin treatment (p=0.0001). Conclusion: Our results revealed that antiapoptotic effects of quercetin, which has predominantly antioxidant effects, may be useful in reducing effects of diabetic complications and preventing new complications.

Keywords:

Experimental diabetic nephropathy, Quercetin, kidney, apoptosis, oxidative stress,

PDF

___

1. Tucker BJ, Collins RC, Ziegler MG, Blantz, RC. Disassociation between glomeruler hyperfiltration and extracellular volume in diabetic rats. Kidney Int. 1991;39:1176-83.
2. King AJ. The use of animal models in diabetes research. Br J Pharmacol. 2012;166:877-94.
3. Tunçdemir M, Ozturk M. The effects of ACE inhibitor and angiotensin receptor blocker on clusterin and apoptosis in the kidney tissue of streptozotocin-diabetic rats. J Mol Histol. 2008;39:605-16.
4. Bolzan AD, Bianchi MS. Genotoxicity of streptozotocin. Mutat Res. 2002;512:121-34.
5. Gao Y, Chen ZY, Wang Y, Liu Y, Ma JX, Li YK. Long non‑coding RNA ASncmtRNA‑2 is upregulated in diabetic kidneys and high glucose‑treated mesangial cells. Exp Ther Med. 2017;13:581-87.
6. Ishikawa Y, Kitamura M. Bioflavonoid quercetin inhibits mitosis and apoptosis of glomerular cells in vitro and in vivo. Biochem Biophys Res Commun. 2000;279:629-34.
7. Jiang XS, Chen XM, Wan JM, Gui HB, Ruan XZ, Du XG. Autophagy Protects against Palmitic Acid-Induced Apoptosis in Podocytes in vitro. Sci Rep. 2017;7:42764.
8. Akef H, Kotb N, Abo-Elmatty D, Salem S. Anti-proliferative Effects of Androctonus amoreuxi Scorpion and Cerastes cerastes Snake Venoms on Human Prostate Cancer Cells. J Cancer Prev. 2017;22:40-6.
9. Kahraman A, Erkasap N, Serteser M, Köken T. Protective effect of quercetin on renal ischemia-reperfusion injury in rats. J Nephrol. 2003;16:219-24.
10. de Souza SR, de Miranda Neto MH, Martins Perles JV, Vieira Frez FC, Zignani I, Ramalho FV, et al. Antioxidant effects of the quercetin in the jejunal myenteric innervation of diabetic rats. Front Med (Lausanne). 2017;7:4-8.
11. Elbe H, Vardi N, Esrefoglu M, Ates B, Yologlu S, Taskapan C. Amelioration of streptozotocin-induced diabetic nephropathy by melatonin, quercetin and resveratrol in rats. Hum Exp Toxicol. 2015;34:100-13.
12. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry. 1979;95:351-58.
13. Sun Y, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clin Chem. 1998;34:497-500.
14. Witko-Sarsat V, Friendlander M, Capeillere-Blandin C. Advanced oxidation protein products as a novel marker of oxidative stress in uremia. Kidney Int. 1996;49:1304-13.
15. Akash MS, Rehman K, Chen S. Spice plant Allium cepa: Dietary supplement for treatment of type 2 diabetes mellitus. Nutrition. 2014;30:1128-137.
16. Anwar MM, Meki AR. Oxidative stress in streptozotocin-induced diabetic rats: effects of garlic oil and melatonin. Comp Biochem Physiol A Mol Integr Physiol. 2003;135:539-47.
17. Shetty AK, Rashmia R, Rajanb MGR, Sambaiaha K, Salimatha PV. Antidiabetic influence of quercetin in streptozotocin induced diabetic rats. Nutr Res. 2004;24:373-81.
18. Anjaneyulu M, Chopra K. Quercetin, an anti-oxidant bioflavonoid, attenuates diabetic nephropathy in rats. Clin Exp Pharmacol Physiol. 2004;31:244-48.
19. Kanter M, Aktas C, Erboga M. Protective effects of quercetin against apoptosis and oxidative stress in streptozotocin-induced diabetic rat testis. Food Chem Toxicol. 2012;50:719-25.
20. Adewole S, Caxton-Martins E, Ojewole J. Protective effect of quercetin on the morphology of pancreatic β-cells of streptozotocin-treated diabetic rats. Afr J Tradit Complement Altern Med. 2006;4:64-74.
21. Arya A, Al-Obaidi MM, Shahid N, Bin Noordin MI, Looi CY, Wong WF, et al. Synergistic effect of quercetin and quinic acid by alleviating structural degeneration in the liver, kidney and pancreas tissues of STZ-induced diabetic rats: A mechanistic study. Food Chem Toxicol. 2014;71:183-96.
22. Dias AS, Porawski M, Alonso M, Marroni N, Collado PS, Gonza´lez-Gallego J. Quercetin decreases oxidative stress, NF-kappaB activation, and iNOS overexpression in liver of streptozotocin-induced diabetic rats. J Nutr. 2005;135:2299-304.
23. Obrosova I, Fathallah L, Liu E, Nourooz-Zadeh J. Early oxidative stress in the diabetic kidney: effect of DL-alpha-lipoic acid. Free Radic Biol Med. 2003;2:186-95.
24. Pan HZ, Zhang L, Guo MY, Sui H, Li H, Wu WH, et al. The oxidative stress status in diabetes mellitus and diabetic nephropathy. Acta Diabetol. 2010;47:71-6.
25. Cumaoglu A, Cevik C, Rackova L, Ari N, Karasu C. Effects of antioxidant stobadine on protein carbonylation, advanced oxidation protein products and reductive capacity of liver in streptozotocin-diabetic rats: role of oxidative/nitrosative stress. Biofactors. 2007;30:171-8.
26. Sefi M, Fetoui H, Makni M, Zeghal N. Mitigating effects of antioxidant properties of Artemisia campestris leaf extract on hyperlipidemia, advanced glycation end products and oxidative stress in alloxan-induced diabetic rats. Food Chem Toxicol. 2010;48:1986-93.
27. Shi XY, Hou FF, Niu HX, Wang GB, Xie D, Guo ZJ, et al. Advanced oxidation protein products promote inflammation in diabetic kidney through activation of renal nicotinamide adenine dinucleotide phosphate oxidase. Endocrinology. 2008;149:1829-39.
28. Zhang G, Oldroyd SD, Huang LH, Yang B, Li Y, Ye R, et al. Role of apoptosis and Bcl-2/Bax in the development of tubulointerstitial fibrosis during experimental obstructive nephropathy. Exp Nephrol. 2001;9:71-80.
29. Kim H, Moon JY, Ahn KS, Cho SK. Quercetin induces mitochondrial mediated apoptosis and protective autophagy in human glioblastoma U373MG cells. Oxid Med Cell Longev. 2013;2013:596496.
30. Liang W, Li X, Li C, Liao L, Gao B, Gan H, et al. Quercetin-mediated apoptosis via activation of the mitochondrial-dependent pathway in MG-63 osteosarcoma cells. Mol Med Rep. 2011;4:1017-23.