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Relationship of leptin administration with production of reactive oxygen species, sperm DNA fragmentation, sperm parameters and hormone profile in the adult rat

  • Reproductive Medicine
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Abstract

Purpose

Leptin, an adipose tissue-derived hormone, plays an important role in energy homeostasis and metabolism, and in the neuroendocrine and reproductive systems. The function of leptin in male reproduction is unclear; however, it is known to affect sex hormones, sperm motility and its parameters. Leptin induces mitochondrial superoxide production in aortic endothelia and may increase oxidative stress and abnormal sperm production in leptin-treated rats. This study aims to evaluate whether exogenous leptin affects sperm parameters, hormone profiles, and the production of reactive oxygen species (ROS) in adult rats.

Methods

A total of 65 Sprague–Dawley rats were divided into three treated groups and a control group. Treated rats received daily intraperitoneal injections of 5, 10 and 30 μg/kg of leptin administered for a duration of 7, 15, and 42 days. Control rats were given 0.1 mL of 0.9 % normal saline for the same period. One day after final drug administration, we evaluated serum specimens for follicle-stimulating hormone (FSH), leutinizing hormone (LH), free testosterone (FT), and total testosterone (TT) levels. Samples from the rat epididymis were also evaluated for sperm parameters and motility characteristics by a Computer-Aided Semen Analysis (CASA) system. Samples were treated with 2′,7′-dichlorofluorescein-diacetate (DCFH-DA) and analyzed using flow cytometry and TUNEL to determine the impact of leptin administration on sperm DNA fragmentation.

Results

According to CASA, significant differences in all sperm parameters in leptin-treated rats and their age-matched controls were detected, except for TM, ALH and BCF. Serum FSH and LH levels were significantly higher in rats that received 10 and 30 μg/kg of leptin compared to those treated with 5 μg/kg of leptin in the same group and control rats (P < 0.05). ROS and sperm DNA fragmentation was significantly higher in rats injected with 10 and 30 μg/kg of leptin for 7 and 15 days compared with rats treated with 5 μg/kg of leptin and the control group (P < 0.05) for the same time period. However, at day 42 of treatment, ROS and sperm DNA fragmentation levels significantly decreased in all groups (P < 0.05).

Conclusion

According to these results, leptin can possibly affect male infertility by ROS induction or hormone profile modulation.

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References

  1. Zhang F, Basinski MB, Beals JM, Briggs SL, Churgay LM, Clawson DK, DiMarchi RD, Furman TC, Hale JF, Hsiung HM, Schoner BE, Smith DP, Zhang XY, Wery JP, Schevitz RW (1997) Crystal structure of the obese protein leptin–E100. Nature 387:206–209

    Article  PubMed  CAS  Google Scholar 

  2. Dallongeville J, Fruchart JC, Auwerx J (1998) Leptin, a pleiotropic hormone: physiology, pharmacology, and strategies for discovery of leptin modulators. J Med Chem 41(27):5337–5352

    Article  PubMed  CAS  Google Scholar 

  3. Cunningham MJ, Clifton DK, Steiner RA (1999) Leptin’s actions on the reproductive axis: perspectives and mechanisms. Biol Reprod 60(2):216–222

    Article  PubMed  CAS  Google Scholar 

  4. Quinton ND, Smith RF, Clayton PE, Gill MS, Shalet S, Justice SK (1999) Leptin binding activity changeswith age: the link between leptin and puberty. Clin Endocrinol Metab 84:2336–2341

    Article  CAS  Google Scholar 

  5. Wauters M, Considine RV, van Gaal LF (2000) Humanleptin: from adipocyte hormone to an endocrine mediator. Eur J Endocrinol 143:293–311

    Article  PubMed  CAS  Google Scholar 

  6. Tena-Sempere M, Barreiro ML, Lage M, Dieguez C, Casanueva FF (2007) Role of leptin and ghrelin in the regulation of gonadal function. Expert Rev Endocrinol Metab 2:239–249

    Article  CAS  Google Scholar 

  7. Camina JP, Lage M, Menendez C, Grana M, Garcia-Devesa J, Dieguez C, Casanueva FF (2002) Evidence of free leptin in human seminal plasma. Endocrine 17:169–174

    Article  PubMed  CAS  Google Scholar 

  8. Tartaglia LA, Dembski M, Weng X, Deng N, Culpepper J, Devos R, Richards GJ, Campfield LA, Clark FT, Deeds J, Muir C, Sanker S, Moriarty A, Moore KJ, Smutko JS, Mays GG, Woolf EA, Monroe CA, Tepper RI (1995) Identification and expression cloning of a leptin receptor, OB-R. Cell 83:1263–1271

    Article  PubMed  CAS  Google Scholar 

  9. Tena-Sempere M, Manna PR, Zhang FP, Pinilla L, González LC, Diéguez C, Huhtaniemi I, Aguilar E (2001) Molecular mechanisms of leptin action in adult rat testis: potential targets for leptin-induced inhibition of steroidogenesis and pattern of leptin receptor messenger ribonucleic acid expression. J Endocrinol 170(2):413–423

    Article  PubMed  CAS  Google Scholar 

  10. De Ambrogi M, Spinaci M, Galeati G, Tamanini C (2007) Leptin receptor in boar spermatozoa. Int J Androl 30(5):458–561

    Article  PubMed  Google Scholar 

  11. Soyupek S, Armağan A, Serel TA, Hoşcan MB, Perk H, Karaöz E, Candir O (2005) Leptin expression in the testicular tissue of fertile and infertile men. Arch Androl 51(3):239–246

    Article  PubMed  CAS  Google Scholar 

  12. Jope T, Lammert A, Kratzsch J, Paasch U, Glander HJ (2003) Leptin and leptin receptor in human seminal plasma and in human spermatozoa. Int J Androl 26:335–341

    Article  PubMed  CAS  Google Scholar 

  13. Caprio M, Isidori AM, Carta AR, Moretti C, Dufan ML, Fabbri A (1999) Expression of functional leptin receptors in rodent Leydig cells. Endocrinology 140:4939–4947

    Article  PubMed  CAS  Google Scholar 

  14. Aquila S, Gentile M, Middea M, Catalano S, Morelli C, Pezzi V et al (2005) Leptin secretion by human ejaculated spermatozoa. J Clin Endocrinol Metab 90:4753–4761

    Article  PubMed  CAS  Google Scholar 

  15. Lackey BR, Gray SL, Henricks DM (2002) Measurement of leptin and insulin-like growth factor-I in seminal plasma from different species. Physiol Res 51:309–311

    PubMed  CAS  Google Scholar 

  16. Yu WH, Kimura M, Walczewska A, Karanth S, Mccann SM (1997) Role of leptin in hypothalamic-pituitary function. Proc Natl Acad Sci 94:1023–1028

    Article  PubMed  CAS  Google Scholar 

  17. Mah PM, Wittert GA (2010) Obesity and testicular function. Mol Cell Endocrinol 316:180–186

    Article  PubMed  CAS  Google Scholar 

  18. Banks WA, Mclay RN, Kastin AJ, Sarmicnto U, Scully S (1999) Passage of leptin across the blood-testis barrier. Am J Physiol 276:1099–1104

    Google Scholar 

  19. Ishikawa T, Fujioka H, Ishimura T (2007) Expression of leptin and leptin receptor in the testis of fertile and infertile patients. Andrologia 39:22–27

    Article  PubMed  CAS  Google Scholar 

  20. El-Hefnawy T, Ioffe S, Dym M (2000) Expression of the leptin receptor during germ cell development in the mouse testis. Endocrinology 141:2624–2630

    Article  PubMed  CAS  Google Scholar 

  21. Frisch RE, McArthur JW (1974) Menstrual cycles: fatness as a determinant of minimum weight for height necessary for their maintenance or onset. Science 185:949–951

    Article  PubMed  CAS  Google Scholar 

  22. Ronnekleiv OK, Ojeda SR, McCann SM (1978) Undernutrition, puberty and the development of estrogen positive feedback in the female rat. Biol Reprod 19:414–424

    Article  PubMed  CAS  Google Scholar 

  23. Yu WH, Kimura M, Walczewska A, Karanth S, Mccann SM (1997) Role of leptin in hypothalamic–pituitary function. Physiology 94:1023–1028

    CAS  Google Scholar 

  24. Tena-Sempere M, Pinilla L, Zhang FP, Gonzales LC, Huhtaniemi I, Casanueva FF, Dieguez C, Aguilar E (2001) Developmental and hormonal regulation of leptin receptor (Ob-R) messenger ribonucleic acid expression in rat testis. Biol Reprod 64:634–643

    Article  PubMed  CAS  Google Scholar 

  25. Glander HJ, Lammert A, Paasch U, Glasow A, Kratzsch J (2002) Leptin exists in tubuli seminiferi and in seminal plasma. Andrologia 34:1–7

    Article  Google Scholar 

  26. Glander HJ, Kratzsch J (2000) Progressive motility of human spermatozoa is negatively correlated with leptin concentration in seminal plasma but not with leptin in serum. Hum Reprod 15:108

    Google Scholar 

  27. Aquila S, Gentile M, Middea M, Catalano S, Morelli C, Pezzi V (2005) Leptin secretion by human ejaculated spermatozoa. J Clin Endocrinol Metab 90:4753–4761

    Article  PubMed  CAS  Google Scholar 

  28. Chebab FF, Mounzih K, Lu R, Lim ME (1997) Early onset of reproductive function in normal female mice treated with leptin. Science 275:88–90

    Article  Google Scholar 

  29. Grosfeld A, Andre J, Hauguel-De Mouzon S, Berra E, Pouyssegur J, Guerre-Millo M (2002) Hypoxia-inducible factor 1 transactivates the human leptin gene promoter. J Biol Chem 277:42953–42957

    Article  PubMed  CAS  Google Scholar 

  30. Lee G, Proenca R, Montez J, Carroll K, Darvishzade J, Lee J, Friedman J (1996) Abnormal splicing of the leptin receptor in diabetic mice. Nature 379:632–635

    Article  PubMed  CAS  Google Scholar 

  31. Glander HJ, Lammert A, Paasch U, Glasow A, Kratzsch J (2002) Lepin exits in tubuli seminiferi and in seminal plasma. Andrologia 34:227–233

    Article  PubMed  CAS  Google Scholar 

  32. Camina JP, Lage M, Menendez C, Grana M, Garcia-Devesa J, Dieguez C (2002) Evidence of free leptin in human seminal plasma. Endocrine 17:169–174

    Article  PubMed  CAS  Google Scholar 

  33. Jope T, Lammert A, Kratzsch J, Paasch U, Glander HJ (2003) Leptin and leptin receptor in human seminal plasma and in human spermatozoa. Int J Androl 26:335–341

    Article  PubMed  CAS  Google Scholar 

  34. Lackey BR, Gray SL, Henricks DM (2002) Measurement of leptin and insulin-like growth factor-I in seminal Plasma from different species. Physiol Res 51:309–311

    PubMed  CAS  Google Scholar 

  35. Aitken RJ, West KM (1990) Analysis of the relationship between reactive oxygen species production and leucocyte infiltration in fractions of human semen separated on Percoll gradients. Int J Androl 13:433–451

    Article  PubMed  CAS  Google Scholar 

  36. Ollero M, Gil-Guzman E, Lopez MC, Sharma RK, Agarwal A, Larson KL (2001) Characterization of subsets of human spermatozoa at different stages of maturation: implications in the diagnosis and treatment of male infertility. Hum Reprod 16:1912–1921

    Article  PubMed  CAS  Google Scholar 

  37. Agarwal A, Prabakaran SA, Said TM (2005) Prevention of oxidative stress injury to sperm. J Androl 26:654–660

    Article  PubMed  CAS  Google Scholar 

  38. Aitken RJ, Baker MA, Sawyer D (2003) Oxidative stress in the male germ line and its role in the aetiology of male infertility and genetic disease. Reprod Biomed Online 7:65–70

    Article  PubMed  CAS  Google Scholar 

  39. Agarwal A, Nandipati KC, Sharma RK, Zippe CD, Raina R (2006) Role of oxidative stress in the pathophysiological mechanism of erectile dysfunction. J Androl 27:335–347

    Article  PubMed  CAS  Google Scholar 

  40. Davi G, Falco A (2005) Oxidant stress, inflammation and atherogenesis. Lupus 14:760–764

    Article  PubMed  CAS  Google Scholar 

  41. Dandona P, Aljada A, Chaudhuri A, Mohanty P, Garg R (2005) Metabolic syndrome: a comprehensive perspective based on interactions between obesity, diabetes, and inflammation. Circulation 111:1448–1454

    Article  PubMed  Google Scholar 

  42. Zak LJ, Cosgrove JR, Aherne FX, Foxcroft GR (1997) Pattern of feed intake and associated metabolic and endocrine changes differentially affect post weaning fertility in primiparous lactating sows. J Anim Sci 75:208–211

    PubMed  CAS  Google Scholar 

  43. Campfield LA, Smith FJ, Burn P (1996) The OB protein (leptin) pathway. A link between adipose tissue mass and central neural networks. Horm Metab Res 28:619–631

    Article  PubMed  CAS  Google Scholar 

  44. Huang L, Cai L (2000) Leptin: a multifunctional hormone. Cell Res 10:81–92

    Article  PubMed  CAS  Google Scholar 

  45. Moschos S, Chan JL, Mantzoros CS (2002) Leptin and reproduction: a review. Fertil Steril 77:433–444

    Article  PubMed  Google Scholar 

  46. Banks WA, McLay R, Kastin AJ, Sarmiento U, Scully S (1999) Passage of leptin across the blood–testis barrier. Am J Physiol Endocrinol Metab 276:1099–1104

    Google Scholar 

  47. Steiman N, Gamzu R, Yogev L, Botchan A, Schreiber L, Yavetz H (2001) Serum leptin concentrations are higher in azoospermic than in normozoospermic men. Fertil Steril 75:821–822

    Article  Google Scholar 

  48. Li HW, Chiu PC, Cheung MP, Yeung WS, WS O (2008) Effect of leptin on motility, capacitation and acrosome reaction of human spermatozoa. Int J Androl 32:687–694

    Article  PubMed  Google Scholar 

  49. Cheung C, Lok S, Weigle D, Finn P, Clifton D, Steiner R (1996) Distribution of leptin receptor(OB-R) mRNA in the central nervous system of the rat. In: 26th Annual Meeting Society for Neuroscience, Washington DC (Abstract)

  50. Barash IA, Cheung CC, Weigle DS, Ren H, Kabigting EB, Kuijper J (1996) Leptin is a metabolic signal to the reproductive system. Endocrinology 137:3144–3147

    Article  PubMed  CAS  Google Scholar 

  51. Haron MN, D’Souza UJ, Jaafar H, Zakaria R, Jeet Singh H (2009) Exogenous leptin administration decreases sperm count and increases the fraction of abnormal sperm in adult rats. Fertil Steril 93:322–324

    Article  PubMed  Google Scholar 

  52. Van Den Saffele JK, Goemaere S, De Bacquer D, Kaufman JM (1999) Serum leptin levels in healthy ageing men: are decreased serum testosterones and increased adiposity in elderly men the consequence of leptin deficiency? Clin Endocrinol (Oxf) 51:81–88

    Article  Google Scholar 

  53. Giagulli VA, Kaufman JM, Vermeulen A (1994) Pathogenesis of the decreased androgen levels in obese men. J Clin Endocrinol Metab 79:97–1000

    Article  Google Scholar 

  54. Wabitsch M, Ballauff A, Holl R, Blum W, Heinze E, Remschmidt H (2001) Serum leptin, gonadotropin and testosterone concentrations in male patients with anorexia nervosa weight gain. J Clin Endocrinol Metab 86:2982–2988

    Article  PubMed  CAS  Google Scholar 

  55. Luukkaa V, Pesonen U, Huhtaniemi I, Lehtonen A, Tilvis R, Tuomilehto J, Koulu M, Huupponen R (1998) Inverse correlation between serum testosterone and leptin in men. J Clin Endocrinol Metab 83:3243–3246

    Article  PubMed  CAS  Google Scholar 

  56. Agarwal A, Makker K, Sharma R (2008) Clinical relevance of oxidative stress in male factor infertility: an update. Am J Reprod Immunol 59:2–11

    Article  PubMed  CAS  Google Scholar 

  57. Aitken RJ, Keith T, Jones Sarah A (2012) Robertson. Reactive oxygen species and sperm function- in sickness and health. J, Androl

    Google Scholar 

  58. Konukoglu D, Serin O, Turhan MS (2006) Plasma leptin and its relationship with lipid peroxidation and nitric oxide in obese female patients with or without hypertension. Arch Med Res 37:602–606

    Article  PubMed  CAS  Google Scholar 

  59. Yamagishi SI, Edelstein D, Du XL (2001) Leptin induces mitochondrial superoxide production and monocyte chemoattractant protein-1 expression in aortic endothelial cells by increasing fatty acid oxidation via protein kinase A. J Biol Chem 276:25096–25100

    Article  PubMed  CAS  Google Scholar 

  60. Shiraishi K, Naito K (2007) Effects of 4-hydroxy-2-nonenal, a marker of oxidative stress, on spermatogenesis and expression of p53 protein in male infertility. J Urol 178:1012–1017

    Article  PubMed  CAS  Google Scholar 

  61. Lackey BR, Gray SL, Henricks DM (2002) Measurement of leptin and insulin-like growth factor-I in seminal plasma from different species. Physiol Res 51:309–311

    PubMed  CAS  Google Scholar 

  62. Chen SC, Kochan JP, Campfield LA, Burn P, Smeyne RJm (1999) 1 Splice variants of the OB receptor gene are differentially expressed in brain and peripheral tissues of mice. J Recep Signal Transduct Res 19:245–266

    Article  CAS  Google Scholar 

  63. Karl PI, Katz R, Daum F, Fisher SE (1991) Mercaptopurine and spermatogenesis in the young rat. Dig Dis Sci 36:1569–1573

    Article  PubMed  CAS  Google Scholar 

  64. Gil-Guzman E, Ollero M, Lopez MC, Sharma RK, Alvarez JG, Thomas AJ Jr et al (2001) Differential production of reactive oxygen species by subsets of human spermatozoa at different stages of maturation. Hum Reprod 16:1922–1930

    Article  PubMed  CAS  Google Scholar 

  65. Gomez E, Irvine DS, Aitkin RJ (1998) Evaluation of a spectrophotometric assay for the measurement of malondialdehyde and 4-hydroxyalkenals in human spermatozoa: relationships with semen quality and sperm function. Int J Androl 21:81–94

    Article  PubMed  CAS  Google Scholar 

  66. World Health Organization (1999) Laboratory manual for the examination of human semen and sperm-cervical mucus interaction, 4th edn. Cambridge University Press, New York

    Google Scholar 

  67. Agarwal A, Said TM (2003) Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum Reprod Update 9:331–345

    Article  PubMed  CAS  Google Scholar 

  68. Von Sobbe HU, Koebnick C, Jenne L, Kiesewetter F (2003) Leptin concentrations in semen are correlated with serum leptin and elevated in hypergonadotrophic hypogonadism. Andrologia 35:233–237

    Article  Google Scholar 

  69. Jorsaraei GhA, Shibahara H, Ayustawati M, HiranoYu Suzuki T, Tahmasbpour Marzony E, Zainalzadeh M, Suzuki M (2010) The Leptin concentrations in seminal plasma of men and its relationship to semen parameters. Iran J Reprod Med 8:95–100

    CAS  Google Scholar 

  70. Li HW, Chiu PC, Cheung MP, Yeung WS, WS O (2008) Effect of leptin on motility, capacitation and acrosome reaction of human spermatozoa. Int J Androl 32:687–694

    Article  PubMed  Google Scholar 

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Acknowledgments

This thesis was funded by a grant from Royan Institute, Tehran, Iran. The authors would like to express their appreciation to Dr. Salmanyazdi, Fariba Soleimaniomid, Ehsan Janzamin and Dr. Elham Amirchaghmaghi for their assistance and advice throughout this project, in addition to their helpful comments on the edition of this article.

Conflict of interest

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Authors and Affiliations

  1. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, 16635-148, Tehran, Iran

    Shima Abbasihormozi, Abdolhossein Shahverdi & Azam Kouhkan

  2. School of Veterinary Medicine, Ilam University, Ilam, Iran

    Shima Abbasihormozi & Javad Cheraghi

  3. Animal core Facility, Reproductive Biomedicine Research Center, Royan Institute for Animal Biotechnology, ACECR, Tehran, Iran

    Abolfazl Kheimeh

  4. Epidemiology and Reproductive Health Department, Reproductive Epidemiology Research Center, Royan Institute for Reproductive Biomedicine, ACECR, 16635-148, Tehran, Iran

    Ali Asghar Akhlaghi

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  1. Shima Abbasihormozi
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  2. Abdolhossein Shahverdi
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  3. Azam Kouhkan
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Correspondence to Azam Kouhkan.

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Abbasihormozi, S., Shahverdi, A., Kouhkan, A. et al. Relationship of leptin administration with production of reactive oxygen species, sperm DNA fragmentation, sperm parameters and hormone profile in the adult rat. Arch Gynecol Obstet 287, 1241–1249 (2013). https://doi.org/10.1007/s00404-012-2675-x

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