Orhan AKMAN, EMİN ŞENGÜL, SEÇKİN ÖZKANLAR, Serpil CAN, AKİF KOÇ, Ali KARADENİZ, Nejdet ŞİMŞEK

The Melatonin Attenuates Alloxan Induced Post-Diabetic Testicular Damage and Oxidative Effects in Rats

Bu çalışmanın amacı, ratlarda alloxan ile oluşturulan diyabet sonrası gözlenen testiküler hasar üzerine melatoninin (MLT) koruyucu etkisini incelemekti. Araştırmada kullanılan 48 adet Sprague Dawley ırkı erkek rat 4 gruba ayrıldı (n=12). Kontrol grubu olan Grup Cye 45 gün süreyle %0.9 serum fizyoloj ik verildi. Grup DMye (Diabetes Mellitus) tek doz alloxan (120 mg/kg) intraperitoneal (IP) olarak enjekte edildi. Grup MLTye 45 gün süreyle 10 mg/kg/gün dozda MLT IP olarak enjekte edildi. Grup DM+MLTye alloxan enjeksiyonunu takiben 45 gün süreyle 10 mg/kg/gün dozda MLT IP olarak uygulandı. Her gruptan 6 hayvana araştırma başladıktan 15 gün sonra, diğer 6 hayvana ise 45 gün sonra ötenazi uygulandı. Kan glikoz seviyesi, oksidan ve antioksidan durumları, sperm parametreleri ve histopatoloj ik ile 8-hydroxy-2′-deoxyguanosine (8-OHdG)-immunopozitif hücre incelemeleri için testis dokusu ve kan örnekleri alındı. Diyabetik ratlara yapılan MLT uygulamasının, grup DMde görülen malondialdehyde (MDA) seviyesi ile 8-OHdG yoğunluğundaki artışı ve glutathion (GSH) ile superoxide dismutase (SOD) seviyelerindeki azalmayı önlediği belirlendi. Alloxan uygulaması grup DMde sperm yoğunluğu, motil sperm sayısı, prematüre akrozom reaksiyonu (AR) ve calcium ionophore A23187 ile indüklenen AR oranlarını deprese ederken, ölü ve/veya anormal sperm oranını değiştirmedi. Ayrıca, histopatoloj ik ve immunohistokimyasal muayeneler, diyabetik ratlara yapılan MLT uygulamasının histoloj ik yapı ve DNA hasarında iyileştirici bir etki gösterdiğini ortaya çıkardı. Sonuç olarak, MLT uygulamasının diyabetik ratlarda oksidatif stresi azaltarak testiküler hasar oluşumunu önlemesine rağmen, sperm parametrelerinde oluşan bozulmaları engelleyemediği gözlendi.

Melatonin Ratlarda Alloxan İle İndüklenen Diyabet Sonrası Oluşan Testiküler Hasarı ve Oksidatif Etkileri Hafifletir

The aim of this study was to examine the protective effect of melatonin (MLT) on the alloxan-induced post-diabetic testicular damage in rats. Forty-eight Sprague Dawley male rats were divided into four groups (n=12). Group C received 0.9% saline for 45 days, as control. Group DM (Diabetes Mellitus) was injected by 120 mg/kg single dose alloxan intraperitoneally (IP). Group MLT was treated by 10 mg/ kg/d MLT IP for 45 d. Group DM+MLT was treated by 10 mg/kg/d MLT IP for 45 days following alloxan injection. Six animals on days 15 and the remaining 6 on days 45 were sacrificed in each group after the initiation of the study. Testis tissues and blood samples were collected to investigate blood glucose, oxidant and antioxidant statues, sperm parameters, and histopathological and 8-hydroxy-2′-deoxyguanosine (8-OHdG)-immunopositive cell examinations. MLT treatment of diabetic rats prevented the increases in malondialdehyde (MDA) levels and 8-OHdG concentrations, and the decreases in the GSH and SOD levels observed in DM groups. Alloxan administration depressed the sperm concentration, the number of progressively motile spermatozoa, premature acrosome reaction (AR) and calcium ionophore A23187-induced AR rates, without any change in dead and/or abnormal sperm rates in DM group. Histopathological and immunohistochemical observations also revealed that MLT-treated diabetic rats had an enhanced histological architecture and DNA damage in testis. These findings suggest that MLT treatment prevents the testicular damage by declining the oxidative stress, but it did not recover the depressed sperm parameters in diabetic rats.

PDF

___

1. WHO: Diabetes: The Cost of Diabetes. Fact Sheet no. 236, 2002.
2. Karunakaran U, Park KG: A systematic review of oxidative stress and safety of antioxidants in diabetes: Focus on islets and their defense. Diabetes Metab J, 37, 106-112, 2013. DOI: 10.4093/dmj.2013.37.2.106
3. Jain GC, Jangir RN: Modulation of diabetes-mellitus-induced male reproductive dysfunctions in experimental animal models with medicinal plants. Pharmacogn Rev, 8, 113-121, 2014. DOI: 10.4103/0973-7847.134245
4. Bal R, Türk G, Tuzcu M, Yilmaz O, Ozercan I, Kuloglu T, Gür S, Nedzvetsky VS, Tykhomyrov AA, Andrievsky GV: Protective effects of nanostructures of hydrated C 60 fullerene on reproductive function in streptozotocin-diabetic male rats. Toxicology, 282, 69-81, 2011. DOI: 10.1016/j.tox.2010.12.003
5. Çayır K, Karadeniz A, Şimşek N, Yıldırım S, Karakuş E, Kara A, Akkoyun HT, Şengül E: Pomegranate seed extract attenuates chemo- therapy-induced acute nephrotoxicity and hepatotoxicity in rats. J Med Food, 14, 1254-1262, 2011. DOI: 10.1089/j mf.2010.0286
6. Karadeniz A, Simsek N, Karakus E, Yildirim S, Kara A, Can I, Kisa F, Emre H, Turkeli M: Royal jelly modulates oxidative stress and apoptosis in liver and kidneys of rats treated with cisplatin. Oxid Med Cell Longev, 2011, 2011. DOI: 10.1155/2011/981793
7. Karakus E, Karadeniz A, Simsek N, Can I, Kara A, Yildirim S, Kalkan Y, Kisa F: Protective effect of Panax ginseng against serum bio- chemical changes and apoptosis in liver of rats treated with carbon tetrachloride (CCl 4). J Hazard Mater, 195, 208-213, 2011. DOI: 10.1016/j. j hazmat.2011.08.027
8. Simsek N, Kaya M, Kara A, Can I, Karadeniz A, Kalkan Y: Effects of melatonin on islet neogenesis and beta cell apoptosis in streptozotocin- induced diabetic rats: an immunohistochemical study. Domest Anim Endocrinol, 43, 47-57, 2012. DOI: 10.1016/j.domaniend.2012.02.002
9. Valavanidis A, V lachogianni T, Fiotakis C: 8-hydroxy-2′ - deoxyguanosine (8-OHdG): A critical biomarker of oxidative stress and carcinogenesis. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev, 27, 120-139, 2009. DOI: 10.1080/10590500902885684
10. Alp BF, Kesik V, Malkoç E, Yigit N, Saldir M, Babacan O, Akgül EÖ, Poyrazoglu Y, Korkmazer N, Gulgun M: The effect of melatonin on procarbazine induced testicular toxicity on rats. Syst Biol Reprod Med, 60, 323-328, 2014. DOI: 10.3109/19396368.2014.930212
11. Armagan A, Uz E, Yilmaz HR, Soyupek S, Oksay T, Ozcelik N: Effects of melatonin on lipid peroxidation and antioxidant enzymes in streptozotocin-induced diabetic rat testis. Asian J Androl, 8, 595-600, 2006. DOI: 10.1111/j.1745-7262.2006.00177.x
12. Guneli E, Tugyan K, Ozturk H, Gumustekin M, Cilaker S, Uysal N: Effect of melatonin on testicular damage in streptozotocin-induced diabetes rats. Eur Surg Res, 40, 354-360, 2008. DOI: 10.1159/000118032
13. Kim S, Lee I, Baek H, Shin I, Moon C, Yun W, Nam K, Kim H, Kim J: Melatonin prevents gentamicin‐induced testicular toxicity and oxidative stress in rats. Andrologia, 46, 1032-1040, 2014. DOI: 10.1111/and.12191
14. Koksal M, Oğuz E, Baba F, Eren MA, Ciftci H, Demir M, Kurcer Z, Take G, Aral F, Ocak A: Effects of melatonin on testis histology, oxidative stress and spermatogenesis after experimental testis ischemia- reperfusion in rats. Eur Rev Med Pharmacol Sci, 16, 582-588, 2012.
15. Baydas G, Canatan H, Turkoglu A: Comparative analysis of the protective effects of melatonin and vitamin E on streptozocin-induced diabetes mellitus. J Pineal Res, 32, 225-230, 2002. DOI: 10.1034/j.1600- 079X.2002.01856.x
16. Champney TH, Holtorf AP, Craft CM, Reiter RJ: Hormonal modulation of pineal melatonin synthesis in rats and Syrian hamsters: Effects of streptozotocin-induced diabetes and insulin injections. Comp Biochem Physiol A Comp Physiol, 83, 391-395, 1986. DOI: 10.1016/0300- 9629(86)90594-3
17. Gurpinar T, Ekerbicer N, Uysal N, Barut T, Tarakci F, Tuglu MI: The histologic evaluation of atorvastatin and melatonin treatment on oxidative stress and apoptosis of diabetic rat pancreas. Kafkas Univ Vet Fak Derg, 16, 547-552, 2010. DOI: 10.9775/kvfd.2009.1122
18. Aksoy N, Vural H, Sabuncu T, Aksoy S: Effects of melatonin on oxidativeantioxidative status of tissues in streptozotocin‐induced diabetic rats. Cell Biochem Funct, 21, 121-125, 2003. DOI: 10.1002/cbf.1006
19. Sudnikovich EJ, Maksimchik YZ, Zabrodskaya SV, Kubyshin VL, Lapshina EA, Bryszewska M, Reiter RJ, Zavodnik IB: Melatonin attenuates metabolic disorders due to streptozotocin-induced diabetes in rats. Eur J Pharmacol, 569, 180-187, 2007. DOI: 10.1016/j.ej phar.2007.05.018
20. Ohkawa H, Ohishi N, Yagi K: Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem, 95, 351-358, 1979. DOI: 10.1016/0003-2697(79)90738-3
21. Tietze F: Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: Applications to mammalian blood and other tissues. Anal Biochem, 27, 502-522, 1969.
22. Sun Y, Oberley LW, Li Y: A simple method for clinical assay of superoxide dismutase. Clin Chem, 34, 497-500, 1988.
23. Akman O, Aksoy M: Assessment of sperm quality by hypo-osmotic swelling test (HOSt) combined by supravital staining in mice. J Anim Res, 18, 1-6, 2008.
24. Türk G, Ateşşahin A, Sönmez M, Yüce A, Çeribaşi AO: Lycopene protects against cyclosporine A-induced testicular toxicity in rats. Theriogenology, 67, 778-785, 2007. DOI: 10.1016/j.theriogenology.2006.10.013
25. Larson JL, Miller DJ: Simple histochemical stain for acrosomes on sperm from several species. Mol Reprod Dev, 52, 445-449, 1999. DOI: 10.1002/(SICI)1098-2795(199904)52:4_____445::AID-MRD14>3.0.CO;2-6
26. Johnsen S: Testicular biopsy score count - A method for registration of spermatogenesis in human testes: Normal values and results in 335 hypogonadal males. Hormones, 1, 2-25, 1970. DOI: 10.1159/000178170
27. Allagui MS, Feriani A, Bouoni Z, Alimi H, Murat JC, El Feki A: Protective effects of vitamins (C and E) and melatonin co-administration on hematological and hepatic functions and oxidative stress in alloxan- induced diabetic rats. J Physiol Biochem, 70, 713-723, 2014. DOI: 10.1007/ s13105-014-0340-5
28. Kadhim HM, Ismail SH, Hussein KI, Bakir IH, Sahib AS, Khalaf BH, Hussain SAR: Effects of melatonin and zinc on lipid profile and renal function in type 2 diabetic patients poorly controlled with metformin. J Pineal Res, 41, 189-193, 2006.
29. She M, Laudon M, Yin W: Melatonin receptors in diabetes: A potential new therapeutical target? Eur J Pharmacol, 744, 220-223, 2014. DOI: 10.1016/j.ej phar.2014.08.012
30. Rochette L, Zeller M, Cottin Y, Vergely C: Diabetes, oxidative stress and therapeutic strategies. Biochim Biophys Acta, 1840, 2709-2729, 2014. DOI: 10.1016/j.bbagen.2014.05.017
31. Islam MS, Loots DT: Experimental rodent models of type 2 diabetes: A review. Methods Find Exp Clin Pharmacol, 31, 249-261, 2009. DOI: 10.1358/mf.2009.31.4.1362513
32. Lenzen S: The mechanisms of alloxan-and streptozotocin-induced diabetes. Diabetologia, 51, 216-226, 2008.
33. Rajendran P, Nandakumar N, Rengarajan T, Palaniswami R, Gnanadhas EN, Lakshminarasaiah U, Gopas J, Nishigaki I: Antioxidants and human diseases. Clin Chim Acta, 436, 332-347, 2014. DOI: 10.1016/j. cca.2014.06.004
34. Walczak-Jedrzejowska R, Wolski JK, Slowikowska-Hilczer J: The role of oxidative stress and antioxidants in male fertility. Cent European J Urol, 66, 60-67, 2013. DOI: 10.5173/cej u.2013.01.art19
35. Ojewale A, Olaniyan O, Faduyile F, Odukanmi O, Oguntola J, Dare B: Testiculo protective effects of ethanolic roots extract of Pseudocedrela kotschyi on alloxan induced testicular damage in diabetic rats. Br J Med Med Res, 4, 548-563, 2014.
36. Shah NA, Khan MR: Antidiabetic effect of Sida cordata in alloxan induced diabetic rats. Biomed Res Int, 2014, 2014. DOI: 10.1155/2014/671294
37. Abbasi Z, Tabatabaei SRF, Mazaheri Y, Barati F, Morovvati H: Effects of sesame oil on the reproductive parameters of diabetes mellitus- induced male rats. World J Mens Health, 31, 141-149, 2013. DOI: 10.5534/ wj mh.2013.31.2.141
38. Nakae D, Akai H, Kishida H, Kusuoka O, Tsutsumi M, Konishi Y: Age and organ dependent spontaneous generation of nuclear 8-hydroxy- deoxyguanosine in male Fischer 344 rats. Lab Invest, 80, 249-261, 2000.
39. Sakai Y, Aminaka M, Takata A, Kudou Y, Yamauchi H, Aizawa Y, Sakagami H: Oxidative stress in mature rat testis and its developmental changes. Dev Growth Differ, 52, 657-663, 2010. DOI: 10.1111/j.1440- 169X.2010.01201.x
40. Singh S, Malini T, Rengarajan S, Balasubramanian K: Impact of experimental diabetes and insulin replacement on epididymal secretory products and sperm maturation in albino rats. J Cell Biochem, 108, 1094- 1101, 2009. DOI: 10.1002/jcb.22337
41. Khaki A, Fathiazad F, Nouri M, Khaki A, Maleki NA, Khamnei HJ, Ahmadi P: Beneficial effects of quercetin on sperm parameters in streptozotocin-induced diabetic male rats. Phytother Res, 24, 1285-1291, 2010. DOI: 10.1002/ptr.3100
42. Suresh S, Prakash S: Effect of Mucuna pruriens (Linn.) on sexual behavior and sperm parameters in streptozotocin-induced diabetic male rat. J Sex Med, 9, 3066-3078, 2012. DOI: 10.1111/j.1743-6109.2010.01831.x
43. Navarro‐Casado L, Juncos-Tobarra M, Cháfer-Rudilla M, Onzoño LÍ, Blázquez-Cabrera J, Miralles-García J: Effect of experimental diabetes and STZ on male fertility capacity. Study in rats. J Androl, 31, 584-592, 2010. DOI: 10.2164/jandrol.108.007260
44. Shrilatha B, Muralidhara: Early oxidative stress in testis and epididymal sperm in streptozotocin-induced diabetic mice: Its progression and genotoxic consequences. Reprod Toxicol, 23, 578-587, 2007. DOI: 10.1016/j.reprotox.2007.02.001
45. 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, 28, 342-347, 2000. DOI: 10.1007/s002400000134
46. Ghosh D, Das UB, Misro M: Protective role of α-tocopherol-succinate (provitamin-E) in cyclophosphamide induced testicular gametogenic and steroidogenic disorders: A correlative approach to oxidative stress. Free Radic Res, 36, 1209-1218, 2002.
47. Scarano W, Messias A, Oliva S, Klinefelter G, Kempinas W: Sexual behaviour, sperm quantity and quality after short‐term streptozotocin‐ induced hyperglycaemia in rats. Int J Androl, 29, 482-488, 2006. DOI: 10.1111/j.1365-2605.2006.00682.x
48. Tanaka M, Nakaya S, Kumai T, Watanabe M, Matsumoto N, Kobayashi S: Impaired testicular function in rats with diet-induced hypercholesterolemia and/or streptozotocin-induced diabetes mellitus. Endocr Res, 27, 109-117, 2001.
49. Hsu PC, Liu MY, Hsu CC, Chen LY, Guo YL: Effects of vitamin E and/ or C on reactive oxygen species-related lead toxicity in the rat sperm. Toxicology, 128, 169-179, 1998. DOI: 10.1016/S0300-483X(98)00068-7
50. Cummins JM, Wakayama T, Yanagimachi R: Fate of microinjected sperm components in the mouse oocyte and embryo. Zygote, 5, 301- 308, 1997. DOI: 10.1017/S0967199400003889
51. Lopes S, Jurisicova A, Sun JG, Casper RF: Reactive oxygen species: Potential cause for DNA fragmentation in human spermatozoa. Hum Reprod, 13, 896-900, 1998. DOI: 10.1093/humrep/13.4.896
52. Roberts S: Book Veterinary Obstetrics and Genital Diseases (Theriogenology), 2nd ed., 726-755, Self Published, Ithaca, New York, 1971.
53. Aitken RJ, Nixon B: Sperm capacitation: A distant landscape glimpsed but unexplored. Mol Hum Reprod, 19, 785-793, 2013. DOI: 10.1093/molehr/gat067
54. Go K, Wolf D: Albumin-mediated changes in sperm sterol content during capacitation. Biol Reprod, 32, 145-153, 1985. DOI: 10.1095/ biolreprod32.1.145
55. Serin İ, Aksoy M, Ceylan A: Cholesterol-loaded cyclodextrin inhibits premature acrosomal reactions in liquid-stored rabbit spermatozoa. Anim Reprod Sci, 123, 106-111, 2011. DOI: 10.1016/j . anireprosci.2010.10.014
56. Lombardo F, Sansone A, Romanelli F, Paoli D, Gandini L, Lenzi A: The role of antioxidant therapy in the treatment of male infertility: An overview. Asian J Androl, 13, 690-697, 2011. DOI: 10.1038/aja.2010.183
57. Comhaire F, Christophe A, Zalata A, Dhooge W, Mahmoud A, Depuydt C: The effects of combined conventional treatment, oral antioxidants and essential fatty acids on sperm biology in subfertile men. Prostaglandins Leukot Essent Fatty Acids, 63, 159-165, 2000. DOI: 10.1054/plef.2000.0174
58. Griveau J, Le Lannou D: Effects of antioxidants on human sperm preparationtechniques. Int J Androl, 17, 225-231, 1994. DOI: 10.1111/ j.1365-2605.1994.tb01247.x
59. Wheeler MB, Seidel GE: Zona pellucida penetration assay for capacitation of bovine sperm. Gamete Res, 18, 237-250, 1987. DOI: 10.1002/mrd.1120180305
60. Kurcer Z, Hekimoglu A, Aral F, Baba F, Sahna E: Effect of melatonin on epididymal sperm quality after testicular ischemia/reperfusion in rats. Fertil Steril, 93, 1545-1549, 2010. DOI: 10.1016/j.fertnstert.2009.01.146
61. Lee K, Lee I, Kim S, Moon C, Park S, Shin D, Park S, Kim H, Kim J: Melatonin attenuates doxorubicin‐induced testicular toxicity in rats. Andrologia, 44, 796-803, 2012. DOI: 10.1111/j.1439-0272.2011.01269.x
62. Sönmez M, Yüce A, Türk G: The protective effects of melatonin and vitamin E on antioxidant enzyme activities and epididymal sperm characteristics of homocysteine treated male rats. Reprod Toxicol, 23, 226- 231, 2007.
63. Wu HJ, Liu C, Duan WX, Xu SC, He MD, Chen CH, Wang Y, Zhou Z, Yu ZP, Zhang L: Melatonin ameliorates bisphenol A-induced DNA damage in the germ cells of adult male rats. Mutat Res, 752, 57-67, 2013. DOI: 10.1016/j.mrgentox.2013.01.005
64. Gwayi N, Bernard R: The effects of melatonin on sperm motility in vitro in Wistar rats. Andrologia, 34, 391-396, 2002. DOI: 10.1046/ j.1439-0272.2002.00522.x