Diyabetik Sıçanların Testis Dokusunda VEGF ve Bcl - 2 Ekspresyon Düzeylerinin İmmünohistokimya ve Western Blot Yöntemleri İle İncelenmes

Year 2016, Volume: 43 Issue: 4, 527 - 533, 25.12.2016

Sevgi İrtegün Engin Deveci

Abstract

Amaç: Bu çalışmada diyabetik sıçanların tetis
dokusunda anjiogenezi ve vasküler geçirgenliği stimule eden vasküler
endotel büyüme faktörünün (VEGF) ve apoptozis regülasyonunda önemli bir
rol oynayan B-cell lymphoma-2’ nin (Bcl-2) ekspresyon düzeylerini
araştırmayı amaçladık.

Yöntemler: Bu çalışmada 20 adet erişkin Wistar albino
sıçan kullanıldı. Sıçanlar kontrol ve Diabetes mellitus (DM) olmak üzere
2 eşit gruba ayrıldı. DM oluşturmak için sitrat tamponunda çözülmüş tek
doz Streptozotosin (STZ) (55mg/kg) intraperitoneal olarak verildi. %10’
luk formaldehit solüsyonuna atılan testis dokuları rutin parafin
takiplerinden sonra histopatolojik olarak incelendi. VEGF ve Bcl-2
protein düzeyleri immünohistokimya ve Western Blot yöntemleriyle
ölçüldü.

Bulgular: Diyabetik gruba ait spermatik hücrelerin
bazılarında dejenerasyon, çekirdekte küçülme ve yer yer piknosis
gözlendi. Diyabet sonucu Sertoli hücrelerinde yapısal değişiklikler
saptanırken, tubuller arasındaki damarlarda dilatasyon ve hemoraji
gözlendi. Diyabetin etkisi ile testis dokusunda VEGF ekspresyonunun
bloke olduğu, Bcl-2 ekspresyonunun ise azaldığı saptandı.

Sonuç: Diyabetik testis dokularında VEGF
ekspresyonunun bloke olmasının yetersiz anjiogenez ve vasküler
permeabiliteye neden olabileceği olasıdır. Ayrıca diyabetik testis
dokularında anti-apoptotik protein olan Bcl-2 ekspresyonundaki azalmanın
diyabetin testiküler dokuda apotozis regülasyonunun bozulmasına neden
olabileceği düşünülmektedir.

Keywords

Diabetes mellitus, testis, VEGF

References

• 1. Mealey BL, Ocampo GL. Diabetes mellitus and periodontal disease. Periodontol 2000 2007;44:127-53.
• 2. American Diabetes Association. Diagnosis and classification of diabetes mellitus. Diabetes Care 2008;31:55-60.
• 3. Zorlu M, Helvacı A, Kıskaç M, et al. Silent myocardial ischemia and related risk factors in patients with type 2 diabetes mellitus. Dicle Med J 2010;37:140-4.
• 4. Cai L, Chen SL, Evans T, et al. Apoptotic germcell death and testicular damage in experimental diabetes: prevention by endothelin antagonism. Uro Res 2000;28:342-7.
• 5. Shrilatha B. Early oxidative stress in testis and epididymal sperm in streptozotocin induced diabetic mice: its progression and enotoxic consequences. Repro Toxicol 2007; 23: 578-587.
• 6. Agbaje IM, Rogers DA, McVicar CM, et al. Insulin dependant diabetes mellitus: implications for male reproductive function. Hum eprod 2007;22:1871-7.
• 7. Kehler DS, Stammers AN, Susser SE, et al. Cardiovascular complications of type 2 diabetes in youth. Biochem Cell Biol 2014;7:1-15.
• 8. Leung DW, Cachianes G, Kuang WJ, et al. Vascular endothelial growth factor is a secreted angiogenic mitogen. Science 1989;246:1306-9.
• 9. Neufeld G, Cohen T, Gengrinovitch S, Poltorak Z. Vascular endothelial growth factor (VEGF) and its receptors. FASEB J 1999;13:9-22.
• 10. Caires KC, de Avila J, McLean DJ. Vascular endothelial growth factor regulates germ cell survival during establishment of permatogenesis in the bovine testis. Reproduction 2009;138:667-77.
• 11. M Kanter, C. Aktas, M. Erboga. Curcumin attenuates testicular damage, apoptotic germ cell death, and oxidative stress in streptozotocin induced diabetic rats. Mol Nutr Food Res 2013;57:1578-85.
• 12. Rashid K, Sil PC. Curcumin ameliorates testicular damage in diabetic rats by suppressing cellular stressmediated mitochondria and endoplasmic reticulum dependent apoptotic death. Biochim Biophys Acta 2015;1852:70-82.
• 13. Jang JH, Surh YJ: Potentiation of cellular antioxidant capacity by Bcl2: implications for its antiapoptotic function. Biochem harmacol 2003;66:1371-9.
• 14. Masiello P, Broca C, GrossR, et al. Experimental NIDDM: development of a new model in adult rats administered streptozotocin and nicotinamide. Diabetes 1998;47:224-9.
• 15. Szkudelski T. The mechanism of alloxan and streptozotocin action in B cells of the rat pancreas. Physiol Res 2001;50:537-46
• 16. 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;11:891-4.
• 17. Öztürk F, Gül M, Ağkadir M, Yağmurca M. Deneysel diyabetin sıçan testislerinde meydana getirdiği histolojik değişiklikler. T Klin J Med Sci 2002;22:173-8.
• 18. Cameron DF, Murray FT, Drylie DD. Interstitial compartment pathology and spermatogenic disruption in testes from impotent diabetic men. Anat Rec 1985;213:53-62.
• 19. Steger RW, Rabe MB. The effect of diabetes mellitus on endocrine and reproductive function. Proc Soc Exp Biol Med 1997;214:1-11.
• 20. Anderson JE, Thliveris JA. Morphometry and cytochemistry of Leydig cells in experimental diabetes. Am J Anat 1987;180:41-8.
• 21. Ebisch IMW, Thomas CMG, Wetzels AMM, et al. Review of the role of the plasminogen activator system and vascular endothelial growth factor in subfertility. Fertil Steril 2008;90:2340-50.
• 22. Reddy N, Kasukurthi KB, Mahla RS, et al. Expression of vascular endothelial growth factor (VEGF) transcript and protein in the testis of several vertebrates, including endangered species. Theriogenology 2012;77:608-14.
• 23. Yoshida A, Anand Apte B, Zetter BR. Differential endothelial migration and proliferation to basic fibroblast growth factor and vascular endothelial growth factor. Growth Factors 1996;13:57-64.
• 24. Sisman AR, Kiray M, Camsari UM, et al. Potential Novel Biomarkers for Diabetic Testicular Damage in Streptozotocin-Induced Diabetic Rats: Nerve Growth Factor Beta and Vascular Endothelial Growth Factor. Disease marker, http://dx.doi.org/10.1155 2014/108106.
• 25. Adler A. Obesity and target organ damage: diabetes. Int J Obes 2002;26:11-14.
• 26. Waisundara VY, Hsu A, Huang D, Tan BKH. Scutellaria baicalensis: enhances the anti diabetic activity of metformin in streptozotocin induced diabeticWistar rats. Am J Chinese Med 2008;36:517-40.
• 27. Long L, Wang J, Lu X, et al. Protective Effects of Scutellarin on Type II Diabetes Mellitus Induced Testicular Damages Rela
• ted to Reactive Oxygen Species/Bcl2/Bax and Reactive Oxygen Species/Microcirculation/Staving Pathway in Diabetic Rat. J Diabetes Res Volume 2015, http://dx.doi.org/10.1155/2015/252530 28. Cai L, Li W, Wang G, et al. Hyperglycemia induced apoptosis in mouse myocardium: mitochondrial cytochrome C-mediated caspase 3 activation pathway. Diabetes 2002;51:1938-48.
• 29. Amin AH, ElMissiry MA, Othman AI. Melatonin ameliorates metabolic risk factors, modulates apoptotic proteins, and protects the rat heart against diabetes induced apoptosis. Eur J pharmacol 2015;747:166-73.