Assessment of Resveratrol's effects comparatively with zinc in experimental rat testicular damage induced by Cyclophosphamide

Cansın Şirin; Canberk Tomruk; Elif Karadadaş; Burcu Denizlioğlu; Gülinnaz Ercan; Aslı Suner; Özlem Yılmaz

doi:10.19161/etd.1201643

EN TR

Assessment of Resveratrol's effects comparatively with zinc in experimental rat testicular damage induced by Cyclophosphamide

Abstract

Aim: To investigate protective effects of Resveratrol in combination or comparison with zinc in experimental testicular injury induced by Cyclophosphamide is studied for the first time in literature. Materials and Methods: Rats (n=63) were randomly divided into 9 groups. After 21 days of drug administration biochemical and histological analysis were performed. Daily water consumption, body weights and weight of testes were measured. Johnsen’s testicular scoring and sperm morphology were evaluated. Hematoxylin&Eosin, Periodic Acid-Schiff and Masson's Trichrome stainings and iNOS, eNOS and CD34 antibodies were applied histologically. To determine oxidative stress, MDA and CAT values were determined. Statistically, one-way ANOVA with post hoc Tukey HSD test for multiple comparisons was performed via IBM SPSS Version 25.0. Results: Cyclophosphamide caused an increase in testicular MDA levels due to elevated oxidant stress. Testicular MDA levels significantly decreased in zinc and Resveratrol groups which revealed protective effects related to Cyclophosphamide treatment, while no significant improvement was observed for control and saline groups. However, the most significant decrease was observed in MDA for Cyclophosphamide+zinc+Resveratrol group in comparison to Cyclophosphamide. Telocytes, which are lately defined novel cells, were detected in the interstitium encircling seminiferous tubules as a sheath immunohistochemically. Conclusion: Not only Resveratrol and zinc, but also their optimum administration separately protects testes in Cyclophosphamide treatment groups. Clinical adaptations of this in vivo model may lead to novel futuristic ideas in preventing infertility due to cancer chemotherapy.

Keywords

Siklofosfamid’in neden olduğu deneysel sıçan testis hasarında Resveratrol’ün etkilerinin çinko ile karşılaştırmalı olarak değerlendirilmesi

Öz

Amaç: Siklofosfamid'in neden olduğu deneysel testis hasarında Resveratrol’ün olası koruyucu etkilerini çinko ile kombine ya da kıyaslamalı olarak araştırılması amaçlanmıştır. Gereç ve Yöntem: Sıçanlar (n=63) rastgele 9 gruba ayrıldı. 21 günlük ilaç uygulaması sonunda; tüm gruplar biyokimyasal ve histolojik olarak incelendi. Günlük su tüketimleri ile deneklerin vücut ağırlıkları ve testis ağırlıkları ölçüldü. Johnsen'in testis skorlaması ve sperm morfolojileri değerlendirildi. Histolojik olarak Hematoksilen&Eozin, Periyodik Asit-Schiff ve Masson'un Trikrom boyaları ve iNOS, eNOS ve CD34 antikorları uygulandı. Doku oksidatif stres düzeyleri için MDA ve CAT değerleri biyokimyasal olarak saptandı. İstatistiksel değerlendirmelerde çoklu karşılaştırmalar için post hoc Tukey HSD testi ile tek yönlü ANOVA, IBM SPSS Sürüm 25.0 kullanıldı. Bulgular: Siklofosfamid, yüksek oksidan stres nedeniyle testis MDA düzeylerini arttırdı. Siklofosfamid tedavisine bağlı koruyucu etkiler ortaya çıkaran çinko ve Resveratrol gruplarında testis MDA seviyelerinde anlamlı düşüşe karşılık, kontrol ve salin gruplarında anlamlı bir iyileşme gözlenmedi. Ancak MDA'da en belirgin azalma Siklofosfamid’e göre Siklofosfamid+çinko+Resveratrol grubunda saptandı. Son zamanlarda yeni tanımlanan hücreler olan telositlerin interstisyumda kılıf şeklinde seminifer tübülleri çevreledikleri immünohistokimyasal olarak gösterildi. Sonuç: Siklofosfamid tedavisi altında testis dokularını korumada sadece Resveratrol ve çinkonun kombine kullanımı değil, optimum dozlarda ayrı ayrı uygulamaları da değerlidir. Bu in vivo modelin klinik uyarlamaları, kanser tedavisine bağlı potansiyel infertilitenin önlenmesinde yeni fikirlerin gelişmesine gelecekte öncülük edebilir.

Anahtar Kelimeler

References

Abraham DJ, Rotella DP, Burger A. Burger’s medicinal chemistry, drug discovery and development. Volume:5 C. Abraham DJ, editor. Wiley-interscience; 2010.
Caglayan C, Temel Y, Kandemir FM, Yildirim S, Kucukler S. Naringin protects against cyclophosphamideinduced hepatotoxicity and nephrotoxicity through modulation of oxidative stress, inflammation, apoptosis, autophagy, and DNA damage. Environmental Science and Pollution Research. 2018;25(21):20968–84.
Nafees S, Rashid S, Ali N, Hasan SK, Sultana S. Rutin ameliorates cyclophosphamide induced oxidative stress and inflammation in Wistar rats: Role of NFκB/MAPK pathway. Chem Biol Interact. 2015;231:98–107.
Oyagbemi AA, Omobowale TO, Saba AB, Adedara IA, Olowu ER, Akinrinde AS, et al. Gallic acid protects against cyclophosphamide-induced toxicity in testis and epididymis of rats. Andrologia. 2016 May 1;48(4):393–401.
Garolla A, Pizzato C, Ferlin A, Carli MO, Selice R, Foresta C. Progress in the development of childhood cancer therapy. Reproductive Toxicology. 2006;22(2):126–32.
Kenney LB, Laufer MR, Grant FD, Grier H, Diller L. High risk of infertility and long term gonadal damage in males treated with high dose cyclophosphamide for sarcoma during childhood. Cancer. 2001;91(3):613–21.
Benzer F, Kandemir FM, Ozkaraca M, Kucukler S, Caglayan C. Curcumin ameliorates doxorubicin-induced cardiotoxicity by abrogation of inflammation, apoptosis, oxidative DNA damage, and protein oxidation in rats. J Biochem Mol Toxicol. 2018;32(2).
Kandemir FM, Kucukler S, Caglayan C, Gur C, Batil AA, Gülçin İ. Therapeutic effects of silymarin and naringin on methotrexate-induced nephrotoxicity in rats: Biochemical evaluation of anti-inflammatory, antiapoptotic, and antiautophagic properties. J Food Biochem. 2017;41(5).

Kaygusuzoglu E, Caglayan C, Kandemir FM, Yıldırım S, Kucukler S, Kılınc MA, et al. Zingerone ameliorates cisplatin‐induced ovarian and uterine toxicity via suppression of sex hormone imbalances, oxidative stress, inflammation and apoptosis in female wistar rats. Biomedicine & Pharmacotherapy. 2018; 102:517–30.
Kuzu M, Kandemir FM, Yildirim S, Kucukler S, Caglayan C, Turk E. Morin attenuates doxorubicin-induced heart and brain damage by reducing oxidative stress, inflammation and apoptosis. Biomedicine and Pharmacotherapy. 2018;106(June):443–53.
Burns J, Yokota T, Ashihara H, Lean MEJ, Crozier A. Plant foods and herbal sources of resveratrol. J Agric Food Chem. 2002;50(11):3337–40.
Levenson AS, Svoboda KM, Pease KM, Kaiser SA, Chen B, Simons LA, et al. Cancer Research. Cancer Res. 2002 Aug 1;58(20):4519–24.
Moussa C, Hebron M, Huang X, Ahn J, Rissman RA, Aisen PS, et al. Resveratrol regulates neuro-inflammation and induces adaptive immunity in Alzheimer’s disease. J Neuroinflammation. 2017;14(1):1–10.
Narayanan BA, Narayanan NK, Re GG, Nixon DW. Differential expression of genes induced by resveratrol in LNCaP cells: P53-mediated molecular targets. Int J Cancer. 2003 Mar 20;104(2):204–12.
Shi T, Liou LS, Sadhukhan P, Duan ZH, Novick AC, Hissong JG, et al. Effects of resveratrol on gene expression in renal cell carcinoma. Cancer Biol Ther. 2004 Sep;3(9):882–8.
Yang S, Kim J, Oh T, Kim M, Lee S, Woo S, et al. Genome-scale analysis of resveratrol-induced gene expression profile in human ovarian cancer cells using a cDNA microarray. Int J Oncol. 2003 Apr 1;22(4):741–50.
Halliwell B. Dietary polyphenols: Good, bad, or indifferent for your health? Cardiovasc Res. 2007;73(2):341–7.
Eleawa SM, Alkhateeb MA, Alhashem FH, Bın-Jalıah I, Sakr HF, Elrefaey HM, et al. Resveratrol Reverses Cadmium Chloride-induced Testicular Damage and Subfertility by Downregulating p53 and Bax and Upregulating Gonadotropins and Bcl-2 gene Expression. Journal of Reproduction and Development. 2014;60(2):115–27.
Juan ME, González-Pons E, Munuera T, Ballester J, Rodríguez-Gil JE, Planas JM. trans-Resveratrol, a natural antioxidant from grapes, increases sperm output in healthy rats. J Nutr. 2005;135(4):757–60.
Shin S, Jeon JH, Park D, Jang MJ, Choi JH, Choi BH, et al. Trans-Resveratrol Relaxes the Corpus Cavernosum Ex Vivo and Enhances Testosterone Levels and Sperm Quality in Vivo. Arch Pharm Res. 2008;31(1):83–7.
Uguralp S, Usta U, Mizrak B. Resveratrol may reduce apoptosis of rat testicular germ cells after experimental testicular torsion. European Journal of Pediatric Surgery. 2005;15(5):333–6.
Yong-guang Jiang, Tao Peng, Yong Luo, Ming-chuan Li, Yun-hua Lin. Resveratrol reestablishes spermatogenesis after testicular injury in rats caused by 2, 5-hexanedione. Chin Med J (Engl). 2008 Jul 1;121(13):1204–9.
Li H, Xia N, Förstermann U. Cardiovascular effects and molecular targets of resveratrol. Vol. 26, Nitric Oxide - Biology and Chemistry. 2012. p. 102–10.
MacDiarmid CW, Taggart J, Jeong J, Kerdsomboon K, Eide DJ. Activation of the Yeast UBI4 Polyubiquitin Gene by Zap1 Transcription Factor via an Intragenic Promoter Is Critical for Zinc-deficient Growth. J Biol Chem. 2016;291(36):18880–96.
Rice JM, Zweifach A, Lynes MA. Metallothionein regulates intracellular zinc signaling during CD4+ T cell activation. BMC Immunol. 2016 Dec 2;17(1):13.
Kambe T, Tsuji T, Hashimoto A, Itsumura N. The Physiological, Biochemical, and Molecular Roles of Zinc Transporters in Zinc Homeostasis and Metabolism. Physiol Rev. 2015 Jul;95(3):749–84.
Cheah Y, Yang W. Functions of essential nutrition for high quality spermatogenesis. Advances in Bioscience and Biotechnology. 2011;02(04):182–97.
Saïd L, Banni M, Kerkeni A, Saïd K, Messaoudi I. Influence of combined treatment with zinc and selenium on cadmium induced testicular pathophysiology in rat. Food and Chemical Toxicology. 2010 Oct;48(10):2759–65.
Pellegrini MSF, Popescu LM. Telocytes. Biomol Concepts. 2011 Jan 1;2(6):481–9.
Hinescu ME, Popescu LM, Gherghiceanu M, Faussone-Pellegrini MS. Interstitial Cajal-like cells in rat mesentery: an ultrastructural and immunohistochemical approach. J Cell Mol Med. 2008;12(1):260–70.
Creţoiu SM, Creţoiu D, Popescu LM. Human myometrium - the ultrastructural 3D network of telocytes. J Cell Mol Med. 2012 Nov;16(11):2844–9.
Lowry OH, Rosebrough NJ, Farr LA, Randall RJ. Protein measurement with the Folin phenol reagent - Journal of Biological Chemistry. 1951;193(1):265–75.
Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979 Jun 1;95(2):351–8.
Yagi K. A simple fluorometric assay for lipoperoxide in blood plasma. Biochem Med. 1976 Apr 1;15(2):212–6.
Aebi H. Catalase in vitro. In: Methods in Enzymology. Academic Press; 1984. p. 121–6.
Latendresse JR, Warbrıttıon lan R, Jonassen H, Creasy DM. Fixation of Testes and Eyes Using a Modified Davidson’s Fluid: Comparison with Bouin’s Fluid and Conventional Davidson’s Fluid. Toxicol Pathol. 2002;30(4):524–533.
Wang Y. Epididymal Sperm Count. In: Current Protocols in Toxicology. Hoboken, NJ, USA: John Wiley & Sons, Inc.; 2003. p. 16.6.1-16.6.5.
Trivedi PP, Tripathi DN, Jena GB. Hesperetin protects testicular toxicity of doxorubicin in rat: Role of NFκB, p38 and caspase-3. Food and Chemical Toxicology. 2011 Apr 1;49(4):838–47.
Johnsen SG. Testicular biopsy score count--a method for registration of spermatogenesis in human testes: normal values and results in 335 hypogonadal males. Hormones. 1970;1(1):2–25.
Irvine DS. Declining sperm quality: a review of facts and hypotheses. Baillieres Clin Obstet Gynaecol. 1997 Dec;11(4):655–71.
Banerjee B, Nandi P, Chakraborty S, Raha S, Sen PC, Jana K. Resveratrol ameliorates benzo(a)pyreneinduced testicular dysfunction and apoptosis: involvement of p38 MAPK/ATF2/iNOS signaling. J Nutr Biochem. 2016;34:17–29.
Cetik S, Ayhanci A, Sahinturk V, Cetik S, Ayhanci A, Sahinturk V. Protective Effect of Carvacrol Against Oxidative Stress and Heart Injury in Cyclophosphamide-Induced Cardiotoxicity in Rat. Brazilian Archives of Biology and Technology. 2015 Aug;58(4):569–76.
El-Sheikh AA, Morsy MA, Okasha AM. Inhibition of NF-κB/TNF-α pathway may be involved in the protective effect of resveratrol against cyclophosphamide-induced multi-organ toxicity. Immunopharmacol Immunotoxicol. 2017 Jul 4;39(4):180–7.
Maremanda KP, Khan S, Jena G. Zinc protects cyclophosphamide-induced testicular damage in rat: Involvement of metallothionein, tesmin and Nrf2. Biochem Biophys Res Commun. 2014 Mar 14;445(3):591–6.
Oboh G, Akomolafe TL, Adetuyi AO. Inhibition of cyclophosphamide-induced oxidative stress in brain by dietary inclusion of red dye extracts from sorghum (Sorghum bicolor) stem. J Med Food. 2010 Oct;13(5):1075–80.
Hosseini A, Zare S, Borzouei Z, Ghaderi Pakdel F, Blvd R. Cyclophosphamide-induced testicular toxicity ameliorate by American ginseng treatment: An experimental study. Vol. 16, Int J Reprod BioMed. 2018. 711-18
Ramos S de P, Goessler KF, Ruiz RJ, Ferrari O, Polito MD, Salles MJS. Exercise protects rat testis from cyclophosphamide-induced damage. Acta Sci Biol Sci. 2013;35(1):105–13.
Nie Z, Zhang L, Chen W, Zhang Y, Hua R, Wang W, et al. The protective effects of pretreatment with resveratrol in cyclophosphamide-induced rat ovarian granulosa cell injury: In vitro study. Reprod Toxicol. 2020 Aug 1;95:66–74.
Zhang Y, Guo L, Law BYK, Liang X, Ma N, Xu G, et al. Resveratrol decreases cell apoptosis through inhibiting DNA damage in bronchial epithelial cells. Int J Mol Med. 2020;45(6):1673–84.
Halsted JA, Ronaghy HA, Abadi P, Haghshenass M, Amirhakemi GH, Barakat RM, et al. Zinc deficiency in man. Am J Med. 1972 Sep 1;53(3):277–84.
Prasad AS, Miale A, Farid Z, Sandstead HH, Schulert AR. Zinc metabolism in patients with the syndrome of iron deficiency anemia, hepatosplenomegaly, dwarfism, and hypognadism. J Lab Clin Med. 1963 Apr;61:537–49.
Kuršvietienė L, Stanevičienė I, Mongirdienė A, Bernatonienė J. Multiplicity of effects and health benefits of resveratrol. Vol. 52, Medicina (Lithuania). Elsevier B.V.; 2016. p. 148–55.
Nie Z, Zhang L, Chen W, Zhang Y, Wang W, Hua R, et al. The protective effects of resveratrol pretreatment in cyclophosphamide-induced rat ovarian injury: an vivo study. Gynecol Endocrinol. 2021;37(10):914–9.
Yeum KJ, Russell RM, Krinsky NI, Aldini G. Biomarkers of antioxidant capacity in the hydrophilic and lipophilic compartments of human plasma. Arch Biochem Biophys. 2004 Oct 1;430(1):97–103.
Yurtçu M, Abasıyanık A, Avunduk MC, Karagözoğlu E, Abasıyanık F. Testis Torsiyonundaki İskemiReperfüzyon Hasarını Önlemede Melatonin ve Steroidin Tek Doz ve Yedi Günlük Tedavilerinin Etkileri. Turkiye Klinikleri Journal of Medical Sciences. 2005;25(4):496–500.
Yuluğ E, Türedi S, Karagüzel E, Kutlu O, Menteşe A, Alver A. The short term effects of resveratrol on ischemiareperfusion injury in rat testis. J Pediatr Surg. 2014 Mar 1;49(3):484–9.
Chen H, Liu J, Luo L, Baig MU, Kim JM, Zirkin BR. Vitamin E, aging and Leydig cell steroidogenesis. Exp Gerontol. 2005 Aug 1;40(8–9):728–36.
Free MJ, Schluntz GA, Jaffe RA. Respiratory Gas Tensions in Tissues and Fluids of the Male Rat Reproductive Tract1. Biol Reprod. 1976 May 1;14(4):481–8.
Peltola V, Mantyla E, Huhtaniemi I, Ahotupa M. Lipid Peroxidation and Antioxidant Enzyme Activities in the Rat Testis after Cigarette Smoke Inhalation or Administration of Polychlorinated Biphenyls or Polychlorinated Naphthalenes. J Androl. 1994 Jul 8;15(4):353–61.
Quinn PG, Payne AH. Oxygen-mediated damage of microsomal cytochrome P-450 enzymes in cultured leydig cells. Role in steroidogenic desensitization. J Biol Chem. 1984 Apr 10;259(7):4130–5.
Zini A, O’Bryan MK, Magid MS, Schlegel PN. Immunohistochemical localization of endothelial nitric oxide synthase in human testis, epididymis, and vas deferens suggests a possible role for nitric oxide in spermatogenesis, sperm maturation, and programmed cell death. Biol Reprod. 1996 Nov;55(5):935–41.
Nair N, Bedwal S, Prasad S, Saini MR, Bedwal RS. Short-term zinc deficiency in diet induces increased oxidative stress in testes and epididymis of rats. Indian J Exp Biol. 2005 Sep;43(9):786–94.
Ghosh D, Das UB, Ghosh S, Mallick M, Debnath J. Testicular Gametogenic and Steroidogenic Activities in Cyclophosphamide Treated Rat: A Correlative Study With Testicular Oxidative Stress. Drug Chem Toxicol. 2002 Jan 7;25(3):281–92.
Zini A, Schlegel PN. Effect of Hormonal Manipulation on mRNA Expression of Antioxidant Enzymes in the Rat Testis. J Urol. 2003 Feb 1;169(2):767–71.
Cheol-hun Son, Dong Yeok Shin, Sung-dae Kim, Hee-seong Park, Min Ho Jung, Jae-ho Bae, et al. Improvement of Antitumor Effect of Intratumoral Injection of Immature Dendritic Cells Into Irradiated Tumor by Cyclophosphamide in Mouse Colon Cancer Model. Journal of Immunotherapy. 2012 Oct 1;35(8):607–14.
Marini M, Rosa I, Guasti D, Gacci M, Sgambati E, Ibba-Manneschi L, et al. Reappraising the microscopic anatomy of human testis: identification of telocyte networks in the peritubular and intertubular stromal space. Sci Rep. 2018 Dec 1;8(1).
Awad M, Ghanem ME. Localization of telocytes in rabbits testis: Histological and immunohistochemical approach. Microsc Res Tech. 2018 Nov 1;81(11):1268–74.
Yifei Liu, Yu Liang, Siyi Wang, Imran Tarique, Waseem Ali Vistro, Haiyan Zhang, et al. Identification and characterization of telocytes in rat testis. Aging. 2019 Aug 14;11(15):5757–68.
Pawlicki P, Hejmej A, Milon A, Lustofin K, Płachno BJ, Tworzydlo W, et al. Telocytes in the mouse testicular interstitium: implications of G-protein-coupled estrogen receptor (GPER) and estrogen-related receptor (ERR) in the regulation of mouse testicular interstitial cells. Protoplasma. 2019 Mar 1;256(2):393–408.
Sidney LE, Branch MJ, Dunphy SE, Dua HS, Hopkinson A. Concise Review: Evidence for CD34 as a Common Marker for Diverse Progenitors. Stem Cells. 2014;32(6):1380.
Abe K, Kameyama H, Abe S ichi. CD34 is Expressed in Endothelial Cells in Embryonic Testes and is Additionally Expressed in Non-Endothelial Cells in Postnatal Mouse Testes. Zoolog Sci. 2022 Jun 9;39(5)

Details

Primary Language

English

Subjects

Health Care Administration

Journal Section

Research Article

Authors

Cansın Şirin
Türkiye

Canberk Tomruk
Türkiye

Elif Karadadaş
Türkiye

Burcu Denizlioğlu
Türkiye

Gülinnaz Ercan
Türkiye

Aslı Suner
Türkiye

Özlem Yılmaz ^*
Türkiye

Publication Date

March 15, 2023

Submission Date

November 9, 2022

Acceptance Date

December 18, 2022

Published in Issue

Year 2023 Volume: 62 Number: 1

DOI

https://doi.org/10.19161/etd.1201643

IZ

https://izlik.org/JA36GU45FY

Cite

RIS / Bibtex

Vancouver

1.Cansın Şirin, Canberk Tomruk, Elif Karadadaş, Burcu Denizlioğlu, Gülinnaz Ercan, Aslı Suner, Özlem Yılmaz. Assessment of Resveratrol’s effects comparatively with zinc in experimental rat testicular damage induced by Cyclophosphamide. EJM. 2023 Mar. 1;62(1):18-29. doi:10.19161/etd.1201643