Medical Journal of Mashhad university of Medical Sciences

Medical Journal of Mashhad university of Medical Sciences

Resveratrol and L-carnitine affects protamine gene expression and sperm parameters in an azoospermic rat model

Document Type : Research Paper

Authors
Hanane Hafezi 1
Akbar Vahdati 1
Mohsen Frozanfar 1
Mehrdad Shariati 2
1 Department of Biology, Shiraz Branch, Islamic Azad University, Shiraz, Iran
2 Department of Biology, Kazerun Branch, Islamic Azad University, Kazerun, Iran
10.22038/mjms.2021.19852
Abstract
The anticancer agent, busulfan, has been reported to adversely affect the male reproductive system such as damaging the sperm parameters. It was shown that L-carnitine and resveratrol can reduce busulfan side effects. This study aimed to evaluate the effects of these agents on the testicular sperm parameters and the expression of the protamine gene in the azoospermic rat model. A total of 78 Sprague Dawley male rats were included in this study and divided into six groups of thirteen rats: control, L-carnitine plus resveratrol (L-ca + RES), busulfan (BUS), busulfan plus L-carnitine (BUS + L-ca), busulfan plus resveratrol (BUS + RES), and busulfan plus L-carnitine plus RES (BUS + L-ca + RES). After 48 days, the sperm parameters and protamine expression were investigated. The results showed significant improvement in the sperm criteria including the increase in sperm motility, viability, count and the expression level of protamine as well as the decrease in abnormal morphology and immature sperm percentage after treatment with L-ca and RES compared to BUS treatment. In conclusion, treatment with L-ca, RES and their combination after administration of BUS may be useful to reduce the side effects of BUS on male fertility by improving the sperm quality. Moreover, our data indicated that the combination of L-ca + BUS + RES was more effective in regulating the cytotoxic effects of busulfan. Hence, we can consider the L-ca and RES as beneficial agents to attenuate the adverse effects of the BUS.
  1. Iwamoto T, Hiraku Y, Oikawa S, Mizutani H, Kojima M, Kawanishi S. DNA intrastrand cross‐link at the 5′‐GA‐3′ sequence formed by busulfan and its role in the cytotoxic effect. Cancer science. 2004;95(5):454-8.
  2. Houot M, Poinsignon V, Mercier L, Valade C, Desmaris R, Lemare F, et al. Physico-chemical stability of busulfan in injectable solutions in various administration packages. Drugs in R&D. 2013;13(1):87-94.
  3. Jung S-W, Kim H-J, Lee B-H, Choi S-H, Kim H-S, Choi Y-K, et al. Effects of Korean Red Ginseng extract on busulfan-induced dysfunction of the male reproductive system. Journal of ginseng research. 2015;39(3):243-9.
  4. Ghasemi M, Bahadori M, Faghani M, Nasiri E, Soleimani Rad J. Buserelin inhibits apoptosis in male germ cells induced by busulfan in mouse testis. J Iran Anat Sci. 2009;7:45-54.
  5. Mertins SD, Myers TG, Holbeck SL, Medina-Perez W, Wang E, Kohlhagen G, et al. In vitro evaluation of dimethane sulfonate analogues with potential alkylating activity and selective renal cell carcinoma cytotoxicity. Molecular cancer therapeutics. 2004;3(7):849-60.
  6. ABIDEMI AA. Protective effect of Tahitian Noni juice on the reproductive functions of Male Wistar Rats traeted with cyclophosphamide. Iranian Journal of Pharmacology and Therapeutics. 2011;10(1):39-0.
  7. Li B, He X, Zhuang M, Niu B, Wu C, Mu H, et al. Melatonin ameliorates busulfan-induced spermatogonial stem cell oxidative apoptosis in mouse testes. Antioxidants & redox signaling. 2018;28(5):385-400.
  8. Tunc O, Thompson J, Tremellen K. Improvement in sperm DNA quality using an oral antioxidant therapy. Reproductive biomedicine online. 2009;18(6):761-8.
  9. De Iuliis GN, Thomson LK, Mitchell LA, Finnie JM, Koppers AJ, Hedges A, et al. DNA damage in human spermatozoa is highly correlated with the efficiency of chromatin remodeling and the formation of 8-hydroxy-2′-deoxyguanosine, a marker of oxidative stress. Biology of reproduction. 2009;81(3):517-24.
  10. Shamsi M, Kumar R, Dada R. Evaluation of nuclear DNA damage in human spermatozoa in men opting for assisted reproduction. Indian Journal of Medical Research. 2008;127(2).
  11. Tanaka H, Baba T. Gene expression in spermiogenesis. Cellular and Molecular Life Sciences CMLS. 2005;62(3):344-54.
  12. Nashtaei MS, Nekoonam S, Naji M, Bakhshalizadeh S, Amidi F. Cryoprotective effect of resveratrol on DNA damage and crucial human sperm messenger RNAs, possibly through 5′ AMP-activated protein kinase activation. Cell and tissue banking. 2018;19(1):87-95.
  13. Agarwal A, Said TM. Carnitines and male infertility. Reproductive biomedicine online. 2004;8(4):376-84.
  14. Ng CM, Blackman MR, Wang C, Swerdloff RS. The role of carnitine in the male reproductive system. Annals of the New York Academy of Sciences. 2004;1033(1):177-88.
  15. Abdelrazik H, Agarwal A. L-carnitine and assisted reproduction. Arch Med Sci. 2009;5:43-7.
  16. Kanter M, Topcu-Tarladacalisir Y, Parlar S. Antiapoptotic effect of L-carnitine on testicular irradiation in rats. Journal of molecular histology. 2010;41(2-3):121-8.
  17. Shin S, Jeon JH, Park D, Jang M-J, Choi JH, Choi B-H, et al. trans-Resveratrol relaxes the corpus cavernosum ex vivo and enhances testosterone levels and sperm quality in vivo. Archives of pharmacal research. 2008;31(1):83-7.
  18. Frémont L. Biological effects of resveratrol. Life sciences. 2000;66(8):663-73.
  19. Juan ME, Gonzalez-Pons E, Munuera T, Ballester J, Rodriguez-Gil JE, Planas JM. trans-Resveratrol, a natural antioxidant from grapes, increases sperm output in healthy rats. The Journal of nutrition. 2005;135(4):757-60.
  20. Turedi S, Yulug E, Alver A, Kutlu O, Kahraman C. Effects of resveratrol on doxorubicin induced testicular damage in rats. Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie. 2015;67(3):229-35.
  21. Jabbari M, Talebi-Taher M, Ariafar M. L-Carnitine and its Protective Role in Contrast-Induced Renal Injury in Rats. Journal of Ardabil University of Medical Sciences. 2012;12(2):115-21.
  22. Karbalay‐Doust S, Noorafshan A, Ardekani FM, Mirkhani H, Baker G. The reversibility of sperm quality after discontinuing nandrolone decanoate in adult male rats. Asian journal of andrology. 2007;9(2):235-9.
  23. Dehghani F, Hassanpour A, Poost-pasand A, Noorafshan A, Karbalay-Doust S. Protective effects of L-carnitine and homogenized testis tissue on the testis and sperm parameters of busulfan-induced infertile male rats. Iranian journal of reproductive medicine. 2013;11(9):693.
  24. Kanatsu-Shinohara M, Toyokuni S, Morimoto T, Matsui S, Honjo T, Shinohara T. Functional assessment of self-renewal activity of male germline stem cells following cytotoxic damage and serial transplantation. Biology of reproduction. 2003;68(5):1801-7.
  25. Anjamrooz SH, Movahedin M, Mowla SJ, Pour Bairanvand S. Assessment of morphological and functional changes in the mouse testis and epididymal sperms following busulfan treatment. Iranian biomedical journal. 2007;11(1):15-22.
  26. Kawashima A, Osman BA, Takashima M, Kikuchi A, Kohchi S, Satoh E, et al. CABS1 is a novel calcium-binding protein specifically expressed in elongate spermatids of mice. Biology of reproduction. 2009;80(6):1293-304.
  27. Turner TT, Lysiak JJ. Oxidative stress: a common factor in testicular dysfunction. Journal of andrology. 2008;29(5):488-98.
  28. Aliabadi E, Mehranjani MS, Borzoei Z, Talaei-Khozani T, Mirkhani H, Tabesh H. Effects of L-carnitine and L-acetyl-carnitine on testicular sperm motility and chromatin quality. Iranian journal of reproductive medicine. 2012;10(2):77.
  29. HOSSEINI AN, Khaki A, Akbari G, GHAFFARI NM. The effect of busulfan on body weight, testis weight and MDA enzymes in male rats. 2014.
  30. Mirhoseini M, Saki G, Hemadi M, Khodadadi A, Asl JM. Melatonin and testicular damage in busulfan treated mice. Iranian Red Crescent Medical Journal. 2014;16(2).
  31. Abd‐Elrazek A, Ahmed‐Farid O. Protective effect of L‐carnitine and L‐arginine against busulfan‐induced oligospermia in adult rat. Andrologia. 2018;50(1):e12806.
  32. Vahdati A, Fathi AR, Nasimi P, Saki G. Busulfan induces apoptotic and cytotoxic effects on testis and epididymal sperm of adult male mouse following low dose treatment. Int J Bio. 2015;5:70-8.
  33. Wu C, Zhang Y, Shen Q, Zhou Z, Liu W, Hua J. Resveratrol changes spermatogonial stem cells (SSCs) activity and ameliorates their loss in busulfan-induced infertile mouse. Oncotarget. 2016;7(50):82085.
  34. Avdatek F, Birdane Y, Türkmen R, Demirel H. Ameliorative effect of resveratrol on testicular oxidative stress, spermatological parameters and DNA damage in glyphosate‐based herbicide‐exposed rats. Andrologia. 2018;50(7):e13036.
  35. Dokmeci D. Oxidative stress, male infertility and the role of carnitines. Folia medica. 2005;47(1):26-30.
  36. Ourique GM, Pês TS, Saccol EM, Finamor IA, Glanzner WG, Baldisserotto B, et al. Resveratrol prevents oxidative damage and loss of sperm motility induced by long-term treatment with valproic acid in Wistar rats. Experimental and Toxicologic Pathology. 2016;68(8):435-43.
  37. AbuMadighem A, Solomon R, Stepanovsky A, Kapelushnik J, Shi Q, Meese E, et al. Development of spermatogenesis in vitro in three-dimensional culture from spermatogonial cells of busulfan-treated immature mice. International journal of molecular sciences. 2018;19(12):3804.
  38. Ni K, Spiess AN, Schuppe HC, Steger K. The impact of sperm protamine deficiency and sperm DNA damage on human male fertility: a systematic review and meta‐analysis. Andrology. 2016;4(5):789-99.
  39. Rogenhofer N, Dansranjavin T, Schorsch M, Spiess A, Wang H, von Schönfeldt V, et al. The sperm protamine mRNA ratio as a clinical parameter to estimate the fertilizing potential of men taking part in an ART programme. Human Reproduction. 2013;28(4):969-78.
  40. Shi J-Z, Zhang S-S, Zhang Z, Liang Q, Shi Y, Hua J-L, et al. Expressions of sperm-specific genes in carnitine-cultured testis sperm of obstructive azoospermia patients. Zhonghua nan ke xue= National journal of andrology. 2010;16(6):504-9.
Medical Journal of Mashhad university of Medical Sciences
Volume 64, Issue 5
March and April 2022
Pages 4186-4195