Antiperoxidative and anti-apoptotic effects of lycopene and ellagic acid on cyclophosphamide-induced testicular lipid peroxidation and apoptosis

Gaffari Türk; Ali Osman Çeribaşi; Fatih Sakin; Mustafa Sönmez; Ahmet Ateşşahin

doi:10.1071/RD09078

RESEARCH ARTICLE

Next Contents Vol 22(4)

Antiperoxidative and anti-apoptotic effects of lycopene and ellagic acid on cyclophosphamide-induced testicular lipid peroxidation and apoptosis

Gaffari Türk ^A ^E , Ali Osman Çeribaşi ^B , Fatih Sakin ^C , Mustafa Sönmez ^A and Ahmet Ateşşahin ^D

+ Author Affiliations

- Author Affiliations

^A Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Fırat University, 23119, Elazığ, Turkey.

^B Department of Pathology, Faculty of Veterinary Medicine, Fırat University, 23119, Elazığ, Turkey.

^C Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Mustafa Kemal University, 31040, Hatay, Turkey.

^D Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Fırat University, 23119, Elazığ, Turkey.

^E Corresponding author. Email: gturk@firat.edu.tr

Reproduction, Fertility and Development 22(4) 587-596 https://doi.org/10.1071/RD09078
Submitted: 28 March 2009 Accepted: 20 October 2009 Published: 9 March 2010

Abstract

The present study was conducted to investigate the possible protective effects of lycopene (LC) and ellagic acid (EA) on cyclophosphamide (CP)-induced testicular and spermatozoal toxicity associated with the oxidative stress and apoptosis in male rats. Forty-eight healthy adult male Sprague-Dawley rats were divided into six groups of eight rats each. The control group was treated with placebo; the LC, EA and CP groups were given LC (10 mg kg^–1), EA (2 mg kg^–1) and CP (15 mg kg^–1), respectively, alone; the CP+LC group was treated with a combination of CP (15 mg kg^–1) and LC (10 mg kg^–1); and the CP+EA group was treated with a combination of CP (15 mg kg^–1) and EA (2 mg kg^–1). All treatments were maintained for 8 weeks. At the end of the treatment period, bodyweight and the weight of the reproductive organs, sperm concentration and motility, testicular tissue lipid peroxidation, anti-oxidant enzyme activity and apoptosis (i.e. Bax and Bcl-2 proteins) were determined. Administration of CP resulted in significant decreases in epididymal sperm concentration and motility and significant increases in malondialdehyde levels. Although CP significantly increased the number of Bax-positive (apoptotic) cells, it had no effect on the number of Bcl-2-positive (anti-apoptotic) cells compared with the control group. However, combined treatment of rats with LC or EA in addition to CP prevented the development of CP-induced lipid peroxidation and sperm and testicular damage. In conclusion, CP-induced lipid peroxidation leads to structural and functional damage, as well as apoptosis, in spermatogenic cells of rats. Both LC and EA protect against the development of these detrimental effects.

Additional keywords: sperm characteristics.

Acknowledgements

The authors acknowledge the financial support of The Scientific and Technological Research Council of Turkey (TÜBİTAK; project no. 106O123. In addition, the authors express their gratitude to Professor Dr Mehmet Çalıcıoğlu (Department of Food Hygiene, Faculty of Veterinary Medicine, Fırat University, Elazığ, Turkey) for help with the English expression.

References

Aebi H. (1983). Catalase. In ‘Methods in Enzymatic Analysis’. (Ed. H. U. Bergmeyer.) pp. 276–286. (Academic Press: New York.)

Agarwal A., Cocuzza M., Abdelrazik H., and Sharma R. K. (2008a). Oxidative stress measurement in patients with male or female factor infertility. In ‘Handbook of Chemiluminescent Methods in Oxidative Stress Assessment’. (Eds I. Popov and G. Lewin.) pp. 195–218. (Transworld Research Network: Kerala.)

Agarwal, A. , Gupta, S. , Sekhon, L. , and Shah, R. (2008b). Redox considerations in female reproductive function and assisted reproduction: from molecular mechanisms to health implications. Antioxid. Redox Signal. 10, 1375–1404.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | Bennett J. P., and Vickery B. H. (1970). Rats and mice. In ‘Reproduction and Breeding Techniques for Laboratory Animals’. (Ed. E. S. E. Hafez.) pp. 299–315. (Lea and Febiger: Philadelphia.)

Cai, L. , Hales, B. F. , and Robaire, B. (1997). Induction of apoptosis in germ cells of adult male rats after exposure to cyclophosphamide. Biol. Reprod. 56, 1490–1497.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | Dollery C. (1999). ‘Therapeutic Drugs.’ (Churchill Livingstone: Edinburgh.)

Elangovan, N. , Chiou, T.-J. , Tzeng, W.-F. , and Chu, S.-T. (2006). Cyclophosphamide treatment causes impairment of sperm and its fertilizing ability in mice. Toxicology 222, 60–70.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | Kandi Coşkun B., and Çobanoğlu B. (2005). Bazal hücreli karsinoma ile skuamoz hücreli karsinomanın histokimyasal özelliklerinin Bax, Bcl-2 ve Ki-67 ile belirlenmesi. Türkderm 39, 185–188. [In Turkish]

Lawrence, R. A. , and Burk, R. F. (1976). Glutathione peroxidase activity in selenium-deficient rat liver. Biochem. Biophys. Res. Commun. 71, 952–958.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Lowry, O. H. , Rosebrough, N. J. , Farr, A. L. , and Randall, R. J. (1951). Protein measurement with folin phenol reagent. J. Biol. Chem. 193, 265–275.
| PubMed | CAS |

Maheshwari, A. , Misro, M. M. , Aggarwal, A. , Sharma, R. K. , and Nandan, D. (2009). Pathways involved in testicular germ cell apoptosis induced by H2O2 in vitro. FEBS J. 276, 870–881.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Matés, J. M. (2000). Effects of antioxidant enzymes in the molecular control of reactive oxygen species toxicology. Toxicology 153, 83–104.
| Crossref | GoogleScholarGoogle Scholar | PubMed |

Pande, M. , and Flora, S. J. S. (2002). Lead induced oxidative damage and its response to combined administration of α-lipoic acid and succimers in rats. Toxicology 177, 187–196.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Papoutsi, Z. , Kassi, E. , Tsiapara, A. , Fokialakis, N. , Chrousos, G. P. , and Moutsatsou, P. (2005). Evaluation of estrogenic/antiestrogenic activity of ellagic acid via the estrogen receptor subtypes ERα and ERβ. J. Agric. Food Chem. 53, 7715–7720.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Pari, L. , and Sivasankari, R. (2008). Effect of ellagic acid on cyclosporine A-induced oxidative damage in the liver of rats. Fundam. Clin. Pharmacol. 22, 395–401.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Placer, Z. A. , Cushman, L. L. , and Johnson, B. C. (1966). Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal. Biochem. 16, 359–364.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Salvemini, F. , Franze, A. , Iervolino, A. , Filosa, S. , Salzano, S. , and Ursini, M. V. (1999). Enhanced glutathione levels and oxidoresistance mediated by increased glucose-6-phosphate dehydrogenase expression. J. Biol. Chem. 274, 2750–2757.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Satoh, K. , Ohyama, K. , Nakagomi, Y. , Ohta, M. , Shimura, Y. , Sano, T. , Ishikawa, H. , Amemiya, S. , and Nakazawa, S. (2002). Effects of growth hormone on testicular dysfunction induced by cyclophosphamide (CP) in GH-deficient rats. Endocr. J. 49, 611–619.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Sedlak, J. , and Lindsay, R. H. (1968). Estimation of total, protein-bound and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal. Biochem. 25, 192–205.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Seeram, N. P. , Adams, L. S. , Henning, S. M. , Niu, Y. , Zhang, Y. , Nair, M. G. , and Heber, D. (2005). In vitro antiproliferative, apoptotic and antioxidant activities of punicalagin, ellagic acid and a total pomegranate tannin extract are enhanced in combination with other polyphenols as found in pomegranate juice. J. Nutr. Biochem. 16, 360–367.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Selvakumar, E. , Prahalathan, C. , Mythili, Y. , and Varalakshmi, P. (2005a). Beneficial effects of dl-α-lipoic acid on cyclophosphamide-induced oxidative stress in mitochondrial fractions of rat testis. Chem. Biol. Interact. 152, 59–66.
| Crossref | dl-α-lipoic acid on cyclophosphamide-induced oxidative stress in mitochondrial fractions of rat testis.&journal=Chem. Biol. Interact.&volume=152&pages=59-66&publication_year=2005&author=E%2E%20Selvakumar&hl=en&doi=10.1016/J.CBI.2005.01.009" target="_blank" rel="nofollow noopener noreferrer" class="reftools">GoogleScholarGoogle Scholar | PubMed | CAS |

Selvakumar, E. , Prahalathan, C. , Mythili, Y. , and Varalakshmi, P. (2005b). Mitigation of oxidative stress in cyclophosphamide-challenged hepatic tissue by dl-α-lipoic acid. Mol. Cell. Biochem. 272, 179–185.
| Crossref | dl-α-lipoic acid.&journal=Mol. Cell. Biochem.&volume=272&pages=179-185&publication_year=2005&author=E%2E%20Selvakumar&hl=en&doi=10.1007/S11010-005-7322-4" target="_blank" rel="nofollow noopener noreferrer" class="reftools">GoogleScholarGoogle Scholar | PubMed | CAS |

Selvakumar, E. , Prahalathan, C. , Sudharsan, P. T. , and Varalakshmi, P. (2006). Chemoprotective effect of lipoic acid against cyclophosphamide-induced changes in the rat sperm. Toxicology 217, 71–78.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Sönmez, M. , Türk, G. , and Yüce, A. (2005). The effect of ascorbic acid supplementation on sperm quality, lipid peroxidation and testosterone levels of male wistar rats. Theriogenology 63, 2063–2072.
| Crossref | GoogleScholarGoogle Scholar | PubMed |

Soong, Y.-Y. , and Barlow, P. J. (2004). Antioxidant activity and phenolic content of selected fruit seeds. Food Chem. 88, 411–417.
| Crossref | GoogleScholarGoogle Scholar | CAS |

Tripathi, D. N. , and Jena, G. B. (2008). Astaxanthin inhibits cytotoxic and genotoxic effects of cyclophosphamide in mice germ cells. Toxicology 248, 96–103.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Türk, G. , Ateşşahin, A. , Sönmez, M. , Yüce, A. , and Çeribaşı, A. O. (2007). Lycopene protects against cyclosporine A-induced testicular toxicity in rats. Theriogenology 67, 778–785.
| Crossref | GoogleScholarGoogle Scholar | PubMed |

Türk, G. , Ateşşahin, A. , Sönmez, M. , Çeribaşı, A. O. , and Yüce, A. (2008). Improvement of cisplatin-induced injuries to sperm quality, the oxidant–antioxidant system, and the histologic structure of the rat testis by ellagic acid. Fertil. Steril. 89(Suppl. 1), 1474–1481.
| Crossref | GoogleScholarGoogle Scholar | PubMed |

Vaisheva, F. , Delbes, G. , Hales, B. F. , and Robaire, B. (2007). Effects of the chemotherapeutic agents for non-Hodgkin lymphoma, cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP), on the male rat reproductive system and progeny outcome. J. Androl. 28, 578–587.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Yılmaz, S. , Ateşşahin, A. , Şahna, E. , Karahan, İ. , and Özer, S. (2006). Protective effect of lycopene on adriamycin-induced cardiotoxicity and nephrotoxicity. Toxicology 218, 164–171.
| Crossref | GoogleScholarGoogle Scholar | PubMed |

Yu, Y.-M. , Chang, W.-C. , Wu, C.-H. , and Chiang, S.-Y. (2005). Reduction of oxidative stress and apoptosis in hyperlipidemic rabbits by ellagic acid. J. Nutr. Biochem. 16, 675–681.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |

Yüce, A. , Ateşşahin, A. , Çeribaşı, A. O. , and Aksakal, M. (2007). Ellagic acid prevents cisplatin-induced oxidative stress in liver and heart tissue of rats. Basic Clin. Pharmacol. Toxicol. 101, 345–349.
| Crossref | GoogleScholarGoogle Scholar | PubMed |

Antiperoxidative and anti-apoptotic effects of lycopene and ellagic acid on cyclophosphamide-induced testicular lipid peroxidation and apoptosis

Abstract

Acknowledgements

Subscriber Login