Tuba YALÇIN, Tuncay KULOĞLU, Gaffari TÜRK, İ Enver OZAN

Orijinal Araştırma Diyabetik Rat Testis Dokusundaki Apoptotik Değişiklikler Üzerine Oleuropeinin Etkisi

Diyabet hastanın yaşam kalitesini azaltarak hayat boyu süren, hastalık yapma ve ölüm oranı yüksek kronik metabolik bir hastalıktır. Bu çalışmada, oksidatif stresin önemli bir rol oynadığını ve streptozotosin STZ ile oluşturulan diyabetik sıçan modelinde Diabetes Mellitus'un DM sıçan testis dokusunda meydana getirdiği apoptotik değişiklikler üzerinde Oleuropein'nin OLE etkisinin incelenmesi amaçlanmıştır. Çalışmada, 28 adet Wistar albino cinsi erkek sıçanlar randomize olarak her grupta 7 hayvan olacak şekilde 4 gruba ayrılarak 6 hafta süren bir deney yapıldı. Kontrol grubundaki sıçanlara herhangi bir uygulama yapılmadı. Diğer 2 DM ve DM + OLE gruba 50 mg/kg olacak şekilde tek doz STZ 0,1 M sodyum sitrat tamponunda çözdürülerek intraperitoneal uygulaması ile oluşturuldu. DM + OLE diyabet oluştuktan sonra ve OLE gruplarına OLE 10ml/kg/gün 6 hafta süreyle oral yolla verildi. Deney sonunda sıçanlar dekapite edilerek testis dokuları çıkarıldı. Testis dokularına Hematoksilen&Eozin, Periyodik Asit Schiff PAS ve TdT-mediated nick and labeling TUNEL boyamaları yapıldı. Testis dokusu örneklerinin bir bölümü ise spermatolojik incelemeler için kullanıldı. Histolojik incelemelerde, DM grubunda tübüllerde atrofi, dejenerasyon peritübüler konjesyon ve bazal membranda kalınlaşama mevcuttu. DM + OLE grubunda ise kontrol grubuna yakın histolojik bulgular gözlendi. TUNEL boyamada, diyabetik grupta apoptotik hücre artışı vardı. Diyabetik grup ile kıyaslandığında DM + OLE grubunda apoptotik hücre artışında anlamlı bir azalma belirlendi. Sadece OLE ugulanan gruba ait sıçan testis dokusunda, interstisyel doku ve seminifer tübüllerde dejenerasyon izlendi. DM’nin oluşturduğu testiküler hasara karşı OLE'nin direkt veya dolaylı antioksidan etkisiyle bu hasarı önlemede etkili olabileceği düşünülmektedir

Anahtar Kelimeler:

Histoloji, Diabetes Mellitus, Oleuropein, Testis

The Effect of Oleuropein on the Apoptotic Changes in the Testicular Tissues of Diabetics Rats

Diabetics is a chronic metabolic condition that proceeds for a lifetime and has a high incidence and mortality rate. In this study, it was aimed to investigate the effects of oleuropein OLE on the apoptotic changes in the testicular tissue caused by Diabetes Mellitus DM in diabetic rat model formed with streptozotocin, in which oxidative stress plays a significant role. In this study, 28 Wistar albino type male rats were randomly divided into 4 groups with each group including 7 rats and the experiment was conducted for 6 months. No implementation was conducted on the control group rats. The other 2 groups DM and DM + OLE were formed by the intraperitoneal implementation, which was implemented by thawing a single dose STZ 0.1 M in sodium citrate buffer in a way that the implementation would be 50mg/kg. OLE administration 10 ml/kg/day was administered orally for DM + OLE after the occurrence of diabetics and OLE groups. As a result of the experiment, the rats were decapitated and the testicular tissues were removed. The testicular tissues were dyed with Hematoxylin&Eosin Periodic Acid Schiff PAS and TdT-mediated nick and labeling TUNEL . Some of the testicular tissue samples were used for spermatological examinations. In the histological examination, it was determined that the DM group had tubular atrophy, degenerated peritubular congestion and thickening in basal membrane. In the DM + OLE group, histological findings close to those of the control group were observed. In the TUNEL dying, an apoptotic cell increase was observed in the diabetic group. Compared to the diabetic group, it was determined that there was a significant decrease in the apoptotic cell increase in the DM + OLE group. In the testicular tissues of the group with sole OLE implementation, interstitial tissue, and degeneration in the seminiferous tubules were observed. It was concluded that against the testicular damage caused by DM, the direct or indirect antioxidant effect of OLE could be effective in preventing that damage

Keywords:

Histology, Diabetes Mellitus, Oleuropei, Testes,

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1. Kağa S. Streptozotozin ile Diyabet Oluşturulan Sıçanlarda Papatya Ekstresinin Antidayabetik ve Antioksidatif Etkisinin Araştırılması. Afyonkarahisar: Afyonkarahisar Kocatepe Üniversitesi, Fen Bilimleri Enstitüsü Yüksek Lisans Tezi 2006.
2. Expert Committee on the Diagnosis and Classification of Diabetes Mellitus. Report of the Expert Commitee on the Diagnosis and Classification of Diabetes. Diabetes Care 2003 Jan; 26 Suppl 1: 5-20.
3. Ricci G, Catizone A, Esposito R, Pisanti FA, Vietri MT, Galdieri M. Diabetic rat testes: morphological and functional alterations. Andrologia 2009; 41: 361-8.
4. Altan N. Diabetes Mellitus ve Oksidatif Stres. Türk Biyokimya Dergisi 2006; 31(2); 51-6
5. Visioli F, Bellosta S, Galli C. Oleuropein, the bitter principles of olives, enhances nitric oxide production by mouse macrophages. Life Sci 1998; 62: 541–6.
6. Carluccio MA, Siculella L, Ancora MA, Massaro M, Scoditti E, Storelli C, Visioli F, Distante A, De Caterina R. Olive oil and red wine antioxidant polyphenols inhibit endothelial activation: antiatherogenic properties of mediterranean diet phytochemicals. Arterioscler Thromb Vasc Biol 2003; 23: 622–9.
7. Owen RW, Giacosa A, Hull WE, Haubner R, Würtele G, Spiegelhalder B, Bartsch H. Olive oil consumption and health: the possible role of antioxidants. Lancet Oncol 2000; 1: 107–12.
8. Tripoli E, Giammanco M, Tabacchi G, Di Majo D, Giammanco S, La Guardia M. The phenolic composition of olive oil: structure, biological activity, and beneficial effects on human health. Nutr Res Rev 2005; 18: 98-112.
9. Fredrickson WR, F and S Group, Inc. Method and Composition for Antiviral Therapy with Olive Leaves. U.S. Patent 2000; 6: 117, 884.
10. Andreadou I, Sigala F, Iliodromitis EK, Papaefthimiou M, Sigalas C, Aligiannis N, Savvari P, Gorgoulis V, Papalabros E,Kremastinos DT. Acute doxorubicin cardiotoxicity is successfully treated with the phytochemical oleuropein through suppression of oxidative and nitrosative stress. J Mol Cell Cardiol 2007; 42: 549–58.
11. Andreadou I, Iliodromitis EK, Mikros E, Constantinou M, Agalias A, Magiatis P, Skaltsounis AL, Kamber E, Tsantili-Kakoulidou A, Kremastinos DT. The olive constituent oleuropein exhibits anti-ischemic, antioxidative, and hypolipidemic effects in anesthetized rabbits. J Nutr 2006; 136: 2213–19.
12. Pitkanen OM, Martin JM, Hallman M, Akerblom HK, Sariola H, Andersson SM. Free radical activity during development of insulindependent diabetes mellitus in the rat. Life Sci 1992; 50: 335 -9.
13. Öztürk F, Ağkadir M, Yağmurca U. Deneysel Diyabetin Sıçan Testislerinde Meydana Getirdiği Histolojik Değişiklikler. T Klin J Med Sci 2002; 22: 173–8.
14. Memişoğulları R. Diyabette Serbest Radikallerin Rolü ve Antioksidanların Etkisi Düzce Tıp Fakültesi Dergisi 2005; 3: 30-9.
15. Al-Azzawie HF, Alhamdani MS. Hypoglycemic and 40. antioxidant effect of oleuropein in alloxan-diabetic rabbits. Life Sci 2006; 78:1371- 7.
16. Bircan FS, Balabanli B, Turkozkan N, Ozan G. Effects of taurine on nitric oxide and 3- nitrotyrosine levels in spleen during endotoxemia. Neurochem Res 2011; 36(11): 1978-83.
17. Ozan G, Turkozkan N, Bircan FS, Balabanli B. Effects of taurine on brain 8- hydroxydeoxyguanosine and3-nitrotyrosine levels in endotoxemia. Neurochem Res 2012; 35(2): 665-70
18. Fox JT, Sakamuru S, Huang R, et al. Mekanisms of antioxidant-induced DNA damage. PNAS 2012; 109(30): E2029.
19. Fox JT, Sakamuru S, Huang R, et al. Highthroughput genotoxicity assay identifies antioxidants as inducers of DNA damage response and cell death. PNAS 2012; 109(14): 5423-28.
20. Dündar Y, Aslan R. Hekimlikte Oksidatif Stres ve Antioksidanlar, 1. Baskı, Ankara 2000; 95.
21. Anderson LC. Effects of alloxan diabetes and insulin in vivo on the ratparotid gland. Am J Physiol 1983; 245: 431-7.
22. Lin SH, Wang ZS. Study on the Expresssion of Androgen Receptor in Testis, Epydidymis and Prostate of Adult Rats with Diabetes. Zhonghua Nan Ke Xue 2005; 11(12); 891-4.
23. Ghafari S, Balajadeh BK, Golalipour MJ. Effect of urtica dioica L. (Urticaceae) on testicular tissue in STZ-induceddibetic rats. Pak. J Biol Sci 2011; 14(16): 798-804.
24. Felter HW, Lloyd JL. 3. King’s American Dispensatory. Cincinnati, OH: Ohio Valley Co 1898.
25. Reiter RJ, Tan DX, Osuna C, Gitto E. Actions of melatonin in the reduction of oxidative stress. A review. J Biomed Sci 2000; 7: 444-58.
26. Cai L, Chen S, Evans T, Deng DX, Mukherjee K, Chakrabarti S. Apoptotic germ-cell death and testicular damage in experimental diabetes: prevention by endothelin antagonism. Urol Res 2000; 28: 342-7.
27. Türk AE. Deneysel Diyabetik Rat Testis Dokusundaki Apoptotik Değişiklikler Üzerine Vitamin D ve Vitamin E’nin Etkisinin Araştırılması. Fırat Üniversitesi Tıp Fakültesi Uzmanlık Tezi 2010.
28. Bal R, Türk G, Tuzcu M, Yılmaz Ö, Özercan İ, Kuloğlu T ve ark. Protective effects of nanostructures of hydrated C60 fullerene on reproductive function in streptozotocin-diabetic male rats. Toxicol 2010; 282 (2011): 69–81.
29. Tsounapi P, Saito M, Dimitriadis F, et al. Antioxidant treatment with edaravone or taurine ameliorates diabetes-induced testicular dysfunction in the rat. Moll Cell Biochem 2012; 369: 195-204.