Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

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

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 http://dx.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.


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.Shah R.2008bRedox considerations in female reproductive function and assisted reproduction: from molecular mechanisms to health implications.Antioxid. Redox Signal.1013751404doi:10.1089/ARS.2007.1964Pubmed Abstract

Agarwal A.Makker K.Sharma R.2008cClinical relevance of oxidative stress in male factor infertility: an update.Am. J. Reprod. Immunol.59211Pubmed Abstract

Aguilar-Mahecha A.Hales B. F.Robaire B.2005Effects of acute and chronic cyclophosphamide treatment on meiotic progression and the induction of DNA double-strand breaks in rat spermatocytes.Biol. Reprod.7212971304doi:10.1095/BIOLREPROD.104.038620Pubmed Abstract

Ahmadi A.Hosseinimehr S. J.Naghshvar F.Hajir E.Ghahremani M.2008Chemoprotective effects of hesperidin against genotoxicity induced by cyclophosphamide in mice bone marrow cells.Arch. Pharm. Res.31794797doi:10.1007/S12272-001-1228-ZPubmed Abstract

Aitken R. J.McLaughlin E. A.2007Molecular mechanisms of sperm capacitation: progesterone-induced secondary calcium oscillations reflect the attainment of a capacitated state.Soc. Reprod. Fertil. Suppl.63273293Pubmed Abstract

Ateşşahin A.Karahan İ.Türk G.Gür S.Yılmaz S.Çeribaşı A. O.2006aProtective role of lycopene on cisplatin-induced changes in sperm characteristics, testicular damage and oxidative stress in rats.Reprod. Toxicol.214247doi:10.1016/J.REPROTOX.2005.05.003Pubmed Abstract

Ateşşahin A.Türk G.Karahan İ.Yılmaz S.Çeribaşı A. O.Bulmuş Ö.2006bLycopene prevents adriamycin-induced testicular toxicity in rats.Fertil. Steril.85Suppl. 112161222doi:10.1016/J.FERTNSTERT.2005.11.035Pubmed Abstract

Ateşşahin A.Çeribaşı A. O.Yüce A.Bulmuş Ö.Çıkım G.2007Role of ellagic acid against cisplatin-induced nephrotoxicity and oxidative stress in rats.Basic Clin. Pharmacol. Toxicol.100121126Pubmed Abstract

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.Robaire B.1997Induction of apoptosis in germ cells of adult male rats after exposure to cyclophosphamide.Biol. Reprod.5614901497doi:10.1095/BIOLREPROD56.6.1490Pubmed Abstract

Chamorro-Cevallos G.Garduno-Siciliano L.Barron B. L.Madrigal-Bujaidar E.Cruz-Vega D. E.Pages N.2008Chemoprotective effect of spirulina (Arthrospira) against cyclophosphamide-induced mutagenicity in mice.Food Chem. Toxicol.46567574doi:10.1016/J.FCT.2007.08.039Pubmed Abstract

Codrington A. M.Hales B. F.Robaire B.2007Chronic cyclophosphamide exposure alters the profile of rat sperm nuclear matrix proteins.Biol. Reprod.77303311doi:10.1095/BIOLREPROD.107.060244Pubmed Abstract

Dollery C. (1999). ‘Therapeutic Drugs.’ (Churchill Livingstone: Edinburgh.)

Elangovan N.Chiou T.-J.Tzeng W.-F.Chu S.-T.2006Cyclophosphamide treatment causes impairment of sperm and its fertilizing ability in mice.Toxicology2226070doi:10.1016/J.TOX.2006.01.027Pubmed Abstract

Flohe L.Otting F.1984Superoxide dismutase assay.Methods Enzymol.19593104doi:10.1016/S0076-6879(84)05013-8

Fukushima T.Yamamoto T.Kikkawa R.Hamada Y.Komiyama M.Mori C.Horii I.2005Effects of male reproductive toxicants on gene expression in rat testes.J. Toxicol. Sci.30195206doi:10.2131/JTS.30.195Pubmed Abstract

Ghosh D.Das U. B.Misro M.2002Protective role of α-tocopherol-succinate (provitamin-E) in cyclophosphamide induced testicular gametogenic and steroidogenic disorders: a correlative approach to oxidative stres.Free Radic. Res.3612091218doi:10.1080/1071576021000016472Pubmed Abstract

Halliwell B.1991Reactive oxygen species in living systems: source, biochemistry and role in human disease.Am. J. Med.91Suppl.S14S22doi:10.1016/0002-9343(91)90279-7Pubmed Abstract

Halliwell B.Gutteridge M. C.1992Biologically relevant metal ion-dependent hydroxyl radical generation.FEBS Lett.307108112doi:10.1016/0014-5793(92)80911-YPubmed Abstract

He D. W.Li X. L.Yue L. Q.Wei G. H.Lin T.Liu J. H.2006Effect of cyclophosphamide on spermatogonial stem cells.Zhonghua Nan Ke Xue12387390, 393Pubmed Abstract

Hou M.Chrysis D.Nurmio M.Parvinen M.Eksborg S.Söder O.Jahnukainen K.2005Doxorubicin induces apoptosis in germ line stem cells in the immature rat testis and amifostine cannot protect against this cytotoxicity.Cancer Res.65999910005doi:10.1158/0008-5472.CAN-05-2004Pubmed Abstract

Howell S. J.Shalet S. M.2005Spermatogenesis after cancer treatment: damage and recovery.J. Natl Cancer Inst. Monogr.20051217doi:10.1093/JNCIMONOGRAPHS/LGI003Pubmed Abstract

İlbey Y. Ö.Özbek E.Şimşek A.Ötünçtemur A.Çekmen M.Somay A.2009Potential chemoprotective effect of melatonin in cyclophosphamide- and cisplatin-induced testicular damage in rats.Fertil. Steril.9211241132doi:10.1016/J.FERTNSTERT.2008.07.1758Pubmed Abstract

Jahnukainen K.Chrysis D.Hou M.Parvinen M.Eksborg S.Söder O.2004Increased apoptosis occurring during the first wave of spermatogenesis is stage-specific and primarily affects midpachytene spermatocytes in the rat testis.Biol. Reprod.70290296doi:10.1095/BIOLREPROD.103.018390Pubmed Abstract

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.Burk R. F.1976Glutathione peroxidase activity in selenium-deficient rat liver.Biochem. Biophys. Res. Commun.71952958doi:10.1016/0006-291X(76)90747-6Pubmed Abstract

Lowry O. H.Rosebrough N. J.Farr A. L.Randall R. J.1951Protein measurement with folin phenol reagent.J. Biol. Chem.193265275Pubmed Abstract

Maheshwari A.Misro M. M.Aggarwal A.Sharma R. K.Nandan D.2009Pathways involved in testicular germ cell apoptosis induced by H2O2 in vitro.FEBS J.276870881doi:10.1111/J.1742-4658.2008.06831.XPubmed Abstract

Matés J. M.2000Effects of antioxidant enzymes in the molecular control of reactive oxygen species toxicology.Toxicology15383104doi:10.1016/S0300-483X(00)00306-1Pubmed Abstract

Pande M.Flora S. J. S.2002Lead induced oxidative damage and its response to combined administration of α-lipoic acid and succimers in rats.Toxicology177187196doi:10.1016/S0300-483X(02)00223-8Pubmed Abstract

Papoutsi Z.Kassi E.Tsiapara A.Fokialakis N.Chrousos G. P.Moutsatsou P.2005Evaluation of estrogenic/antiestrogenic activity of ellagic acid via the estrogen receptor subtypes ERα and ERβ.J. Agric. Food Chem.5377157720doi:10.1021/JF0510539Pubmed Abstract

Pari L.Sivasankari R.2008Effect of ellagic acid on cyclosporine A-induced oxidative damage in the liver of rats.Fundam. Clin. Pharmacol.22395401doi:10.1111/J.1472-8206.2008.00609.XPubmed Abstract

Placer Z. A.Cushman L. L.Johnson B. C.1966Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems.Anal. Biochem.16359364doi:10.1016/0003-2697(66)90167-9Pubmed Abstract

Salvemini F.Franze A.Iervolino A.Filosa S.Salzano S.Ursini M. V.1999Enhanced glutathione levels and oxidoresistance mediated by increased glucose-6-phosphate dehydrogenase expression.J. Biol. Chem.27427502757doi:10.1074/JBC.274.5.2750Pubmed Abstract

Satoh K.Ohyama K.Nakagomi Y.Ohta M.Shimura Y.Sano T.Ishikawa H.Amemiya S.Nakazawa S.2002Effects of growth hormone on testicular dysfunction induced by cyclophosphamide (CP) in GH-deficient rats.Endocr. J.49611619doi:10.1507/ENDOCRJ.49.611Pubmed Abstract

Sedlak J.Lindsay R. H.1968Estimation of total, protein-bound and nonprotein sulfhydryl groups in tissue with Ellman’s reagent.Anal. Biochem.25192205doi:10.1016/0003-2697(68)90092-4Pubmed Abstract

Seeram N. P.Adams L. S.Henning S. M.Niu Y.Zhang Y.Nair M. G.Heber D.2005In 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.16360367doi:10.1016/J.JNUTBIO.2005.01.006Pubmed Abstract

Selvakumar E.Prahalathan C.Mythili Y.Varalakshmi P.2005aBeneficial effects of dl-α-lipoic acid on cyclophosphamide-induced oxidative stress in mitochondrial fractions of rat testis.Chem. Biol. Interact.1525966doi:10.1016/J.CBI.2005.01.009Pubmed Abstract

Selvakumar E.Prahalathan C.Mythili Y.Varalakshmi P.2005bMitigation of oxidative stress in cyclophosphamide-challenged hepatic tissue by dl-α-lipoic acid.Mol. Cell. Biochem.272179185doi:10.1007/S11010-005-7322-4Pubmed Abstract

Selvakumar E.Prahalathan C.Sudharsan P. T.Varalakshmi P.2006Chemoprotective effect of lipoic acid against cyclophosphamide-induced changes in the rat sperm.Toxicology2177178doi:10.1016/J.TOX.2005.08.020Pubmed Abstract

Sinha Hikim A. P.Swerdloff R. S.1999Hormonal and genetic control of germ cell apoptosis in the testis.Rev. Reprod.43847doi:10.1530/ROR.0.0040038Pubmed Abstract

Sönmez M.Türk G.Yüce A.2005The effect of ascorbic acid supplementation on sperm quality, lipid peroxidation and testosterone levels of male wistar rats.Theriogenology6320632072doi:10.1016/J.THERIOGENOLOGY.2004.10.003Pubmed Abstract

Soong Y.-Y.Barlow P. J.2004Antioxidant activity and phenolic content of selected fruit seeds.Food Chem.88411417doi:10.1016/J.FOODCHEM.2004.02.003

Stahl W.Sies H.2003Antioxidant activity of carotenoids.Mol. Aspects Med.24345351doi:10.1016/S0098-2997(03)00030-XPubmed Abstract

Tripathi D. N.Jena G. B.2008Astaxanthin inhibits cytotoxic and genotoxic effects of cyclophosphamide in mice germ cells.Toxicology24896103doi:10.1016/J.TOX.2008.03.015Pubmed Abstract

Türk G.Ateşşahin A.Sönmez M.Yüce A.Çeribaşı A. O.2007Lycopene protects against cyclosporine A-induced testicular toxicity in rats.Theriogenology67778785doi:10.1016/J.THERIOGENOLOGY.2006.10.013Pubmed Abstract

Türk G.Ateşşahin A.Sönmez M.Çeribaşı A. O.Yüce A.2008Improvement of cisplatin-induced injuries to sperm quality, the oxidant–antioxidant system, and the histologic structure of the rat testis by ellagic acid.Fertil. Steril.89Suppl. 114741481doi:10.1016/J.FERTNSTERT.2007.04.059Pubmed Abstract

Vaisheva F.Delbes G.Hales B. F.Robaire B.2007Effects 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.28578587doi:10.2164/JANDROL.106.002428Pubmed Abstract

Yılmaz S.Ateşşahin A.Şahna E.Karahan İ.Özer S.2006Protective effect of lycopene on adriamycin-induced cardiotoxicity and nephrotoxicity.Toxicology218164171doi:10.1016/J.TOX.2005.10.015Pubmed Abstract

Yu Y.-M.Chang W.-C.Wu C.-H.Chiang S.-Y.2005Reduction of oxidative stress and apoptosis in hyperlipidemic rabbits by ellagic acid.J. Nutr. Biochem.16675681doi:10.1016/J.JNUTBIO.2005.03.013Pubmed Abstract

Yüce A.Ateşşahin A.Çeribaşı A. O.Aksakal M.2007Ellagic acid prevents cisplatin-induced oxidative stress in liver and heart tissue of rats.Basic Clin. Pharmacol. Toxicol.101345349doi:10.1111/J.1742-7843.2007.00129.XPubmed Abstract

Zhang X.Yamamoto N.Soramoto S.Takenaka I.2001Cisplatin-induced germ cell apoptosis in mouse testes.Arch. Androl.464349doi:10.1080/01485010150211146Pubmed Abstract



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