Effect of Melatonin and Vitamin C on Damage and Expression of Endothelial NOS in Testes of Chronic Alcoholic Rats

Abstract

The aim of this study was to determine the alterations in testes caused by alcohol consumption and the effects of melatonin and vitamin C on these changes in rats. Twenty-four adult male Wistar rats weighting 200-250g were used in this study. Rats were divided into four groups. The first group served as control (n:6). The second group received ethanol treatment for 28 days (n:6). The third group was given ethanol and supplemented (intraperitoneally) (n:6). The fourth group was given ethanol and supplemented with 4 mg/kg/day melatonin (intraperitoneally) examinations revealed testicular edema and dilatation and congestion of vessels in the ethanol-fed group. In some seminiferous tubules we observed atrophy, desquamation of seminiferous epithelium into the tubular vacuolization. In melatonin and vitamin C treated groups, there was a partial recovery but the damage remained in a great extent. eNOS immunoreactivity was (++) in Leydig cells, (+) in some spermatogenic cells immunoreactivity was increased in degenerated cells. In melatonin and vitamin C treated groups, no significant change in eNOS immunoreactivity was observed. Present results indicated that chronic alcohol consumption caused damage to testicles and especially melatonin and vitamin C administration produced in some degree provided protection against alcoholinduced damage

Keywords

• Chronic alcohol consumption, testes, eNOS, melatonin, vitamin C

MELATONİN VE C VİTAMİNİ’NİN KRONİK ALKOLİK SIÇANLARIN TESTİS DOKUSUNDAKİ HASAR VE eNOS İMMUNREAKTİVİTESİ ÜZERİNE ETKİLERİ

Abstract

Bu çalışmada alkolün testiste oluşturduğu hasar ve eNOS ekspresyonu üzerine melatonin ve C vitamininin koruyucu etkilerinin belirlenmesi amaçlandı. Çalışmada 24 adet 200-250 gr ağırlığında Wistar cinsi ergin erkek sıçan kullanıldı. Sıçanlar rasgele olarak dört gruba ayrıldı. Birinci grup kontrol (n:6); ikinci grup deney süresince (28 gün) alkol içeren sıvı diyet alan (n:6); üçüncü grup deney boyunca alkol içeren sıvı diyet ile birlikte 40mg/kg/gün intraperitoneal C vitamini alan (n:6); dördüncü grup deney süresince alkol içeren sıvı diyet ile birlikte 4mg/kg/gün intraperitoneal melatonin alan sıçanlardan (n:6) oluşturuldu. Deney süresince alkol içeren sıvı diyet alan gruptaki sıçanların testis dokularında ödem, damarlarda dilatasyon ve konjesyon tespit edildi. Bazı seminifer tübüllerde atrofi, spermatogenetik seriye ait hücrelerde lümen içine dökülme, dejenerasyon ve seminifer tübül epitelinde vakuolizasyon gözlendi. Melatonin ve C vitamini uygulanan gruplarda ise nispeten bazı bulgularda düzelme gözlenmesine rağmen hasar devam etmekteydi. eNOS immunoreaktivitesi yapılan boyamada intertisyel Leydig hücrelerinde (++), bazı spermatogenetik seriye ait hücrelerde (+) ve tunika albugineada (+) boyanma ayırt edildi. Dejenere olan hücrelerde immunureaktivitesinde artış saptandı. Melatonin ve C vitamini uygulanan gruplarda boyanmada değişiklik saptanmadı. Sonuç olarak kronik alkol tüketiminin testiste dokusunda hasara yol açtığı, ancak bu hasarın melatonin ve C vitamini verilmesi ile kısmen düzeldiği belirlenmiştir

Keywords

• Kronik alkol tüketimi, testis, eNOS, melatonin, C vitamini

References

1. Maneesh M, Dutta S, Chakrabarti A, Vasudevan DM. Alcohol abuseduration- dependent decrease in plasma testosterone and antioxidants inmales. Indian J Physiol Pharmacol 2006, 50 (3): 291–296.
2. Cicero TJ, Meyer ER, Bell RD. Effects of ethanol on the hypothalamic- pituitary- luteinizing hormone axis and testicular steroidogenesis. Pharmacol Exp Ther 1979; 208: 210–15.
3. Adams ML, Little PJ, Bell B, Cicero TJ. Alcohol affects rat testicular interstitial fl uid volume and testicular secretion of testosterone and betaendorphin. J Pharmacol Exp Ther 1991; 258:1008–14.
4. Giannessi F, Giambelluca MA, Grasso L, Scavuzzo MC, Ruffoli R. Curcumin protects Leydig cells of mice from damage induced by chronic alcohol administration. Med Sci Monit 2008; 14(11):237-42.
5. Quintans LN, Castro GD, Castro JA. Oxidation of ethanol to acetaldehyde and free radicals by rat testicular microsomes. Arch Toxicol 2005; 79:25–30.
6. Lloyd RV, Jin L, Qian X, Zhang S, Scheithauer BW. Nitric oxide synthase in the human pituitary gland. Am J Pathol 1995; 146:86-94.
7. Burnett AL, Ricker DD, Chamness SL, et al. Localization of nitric oxide synthase in the reproductive organs of the male rat. Biol Reprod 1995; 52:1-7.
8. Chamness SL, Ricker DD, Crone JK, et al. The effect of androgen on nitric oxide synthase in the male reproductive tract of the rat. Fertil Steril 1995; 63:111-1107.

9. Ceccatelli S, Hulting AL, Zhang X, et al. Nitric oxide synthase in the rat anterior pituitary gland and the role of nitric oxide in regulation of luteinizing hormone secretion. Proc Natl Acad Sci USA 1993; 90:11292-11296.
10. Reiter RJ, Tan DX, Mayo JC, et al. Pharmacological actions of melatonin in oxygen radical pathophysiology. Acta Biochim Pol 2003; 50:1129–46.
11. Rodriguez C, Mayo JC, Sainz RM, et al. Regulation of antioxidant enzymes: a significant role for melatonin. J Pineal Res 2004; 36:1–9.
12. Ozturk A, Baltacı AK, Mogulkoc R, Ozturk B. The administration on reperfusion damage in experimental Neuro Endocrinol Lett 2003; 24:170–2.
13. Abasiyanik A, Dagdonderen L. Beneficial effects of melatonin compared with allopurinol in experimental testicular torsion. J Ped Surg 2004; 39:1238–41.
14. Duru FI, Noronha CC, Akinwande AI, Okanlawon AO. Effects of torsion, detorsion and melatonin on testicular malondialdehyde level. West Afr J Med 2007, 26:312–5.
15. Halliwell B, Wasil M, Grootveld M. Biologically significant scavenging of the myeloperoxidase-derived hypochlorous acid by ascorbic acid. FEBS Lett 1987; 213:15–17. oxidant
16. Uzbay IT, Kayaalp SO. A modified liquid diet of chronic ethanol administration: validation by ethanol withdrawal syndrome in rats. Phar- macol Res 1995; 31:37–42.
17. Sonmez MF, Colakoglu N, Kukner A, Ozan E, Dabak DO. Immunolocalization of TGF-beta2 in the rat thymus during late stages of prenatal development. Acta Histochem. 2009;111(1):68 -73.
18. Lieber CS. Hepatic and other medical disor- ders of alcoholism: from pathogenesis to treat- ment. J Stud Alcohol 1998; 59:9–25.
19. Herman M, Kang SS, Lee S, James P, Rivier C. Systemic administration of alcohol to adult rats inhibits leydig cell activity: time course of effect and role of nitric oxide. Alcohol Clin Exp Res 2006; 30(9):1479–1491.
20. Gianoulakis C. Characterization of the effects of acute ethanol administration on the release of beta-endorphin peptides by the rat hypo- thalamus. European Journal of Pharmacology 1990; 180:21–29.
21. SIES H. Oxidative stress: Oxidants and antioxidants. Experimental Physiology 1997; 82:291–295.
22. Emanuele NV, Lapaglı N, Steıner J, et al. Peripubertal Endocrine 2001; 14(2): 213–9. Etoh Exposure.
23. Martinez M, Macera S, de Assis GF, et al. Structural evaluation of the effects of chronic ethanol ingestion on the testis of Calomys callosus. Tissue Cell 2009; 41(3):199-205.
24. Gavaler JS, Perez HA, Estes L, Van Thiel DH. Morphologic alterations of rat Leydig cells induced by ethanol. Pharmacol Biochem Behav 1983; 1:341-7.
25. Koh PO, Kim MO. Ethanol exposure decreases cell proliferation and increases apoptosis in rat testes. J Vet Med Sci 2006; 8(10):1013-7.
26. Bamac Y, Colak T, Bamac B, et al. Comparative study on apoptosis in the testes of normal and alcoholic rats. Saudi Med J. 2005; 26(6):928-33.
27. Oner-Iyidoğan Y, Gürdöl F, Oner P. The effects of acute melatonin and ethanol treatment on antioxidant enzyme activities in rat testes. Pharmacol Res 2001; 44(2):89-93.
28. Manev H, Uz T, Joo JY. Increased brain damage after stroke or excitotoxic seizures in melatonin-deficient rats. FASEB J 1996; 10:1546–51.
29. Reiter RJ. Antioxidant actions of melatonin. Adv Pharmacol 1997; 38:103–17.
30. Tan DX, Chen LD, Poeggeler B, Manchester LC, Reiter RJ. Melatonin: a potent endogenous hydroxyl radical scavenger. Endocr J 1993; 1:57–60.
31. Cagnacci A, Volpe A. Influence of melatonin and photoperiod on animal and human reproduction. J Endocrinol Invest 1996; 19:382–411.
32. Yu ZH, Chow PH, Pang SF. Identification and characterization of 2[125I]-iodomelatonin binding sites in the rat epididymis. J Pineal Res 1994; 17:195–201.
33. Ellis CC. Inhibition of rat testicular androgen synthesis in vitro by melatonin and serotonin. Endocrinology 1972; 90:17–28.
34. Ng T, Lo LLH. Inhibitory actions of pineal indoles in steroidogenesis in isolated rat leydig cells. J Pineal Res 1988; 5:229–43.
35. Persengiev S, Kehajova J. Inhibitory action of melatonin and structurally related compounds on testosterone production by mouse Leydig cells in vitro. Cell Biochem Funct 1991; 9:281 –6.
36. Röjdmark S, Wikner J, Adner N, Andersson DEH, Wetterberg L. Inhibition of melatonin secretion by ethanol in man. Metabolism 1993; 42:1047–51.
37. Kurcer Z, Oguz E, Ozbilge H, et al. Effect of melatonin on testicular ischemia/reperfusion injury in rats: is this effect related to the proinflammatory cytokines? Fertil Steril 2008; 89:1468–73.
38. Hsu PC, Liu MY, Hsu CC, Chen LY, Guo YL. Effects of vitamin E and/or C on reactive oxygen toxicity in the rat sperm. Toxicology 1998; 128:169–179.
39. Sen Gupta R, Kim J, Gomes C, et al. Effect of ascorbic acid supplementation on testicular steroidogenesis and germ cell death in cadmium-treated male rats. Mol Cell Endocrinol 2004; 30;221(1-2):57-6.
40. McCann SM, Rettori V. The role of nitric oxide in reproduction. Proc Soc Exp Biol Med 1996; 211:7-15.
41. Rettori V, Belova N, Dees WL, et al. Role of nitric oxide in the control of luteinizing hormone-releasing hormone release in vivo and in vitro. Proc Natl Acad Sci USA 1993; 9010130- 10134.