Comparative ovarian morphophysiology of Wistar rats and Zebrafish after exposure to nandrolone decanoate

João Eudes Farias Cavalcante Filho; Solano Dantas Martins; Jonathan Elias Rodrigues Martins; Jéssica Sales Lobato; Yara Silvino Sales; Sara Rany Alexandre Bittencourt; Fernanda Vitória Almeida Magalhães; Ana Ruth Reinaldo Menezes; Maria Alice Felipe Oliveira; Vânia Marilande Ceccatto; Anderson Weiny Barbalho Silva; Carminda Sandra Brito Salmito-Vanderley; Valdevane Rocha Araújo

doi:10.1590/1984-3143-AR2024-0046

ORIGINAL ARTICLE

Comparative ovarian morphophysiology of Wistar rats and Zebrafish after exposure to nandrolone decanoate

João Eudes Farias Cavalcante Filho; Solano Dantas Martins; Jonathan Elias Rodrigues Martins; Jéssica Sales Lobato; Yara Silvino Sales; Sara Rany Alexandre Bittencourt; Fernanda Vitória Almeida Magalhães; Ana Ruth Reinaldo Menezes; Maria Alice Felipe Oliveira; Vânia Marilande Ceccatto; Anderson Weiny Barbalho Silva; Carminda Sandra Brito Salmito-Vanderley; Valdevane Rocha Araújo

http://dx.doi.org/10.1590/1984-3143-AR2024-0046 Anim Reprod, vol.22, n1, e20240046, 2025

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Abstract

This study aimed to compare the effects of nandrolone decanoate on the morphology and physiology of ovarian tissues in two experimental models, Zebrafish and rats, after in vitro cultivation. A total of 136 animals were used (Wistar rats, n=36, and Zebrafish, n=100). In both experiments, the animals were divided into two groups (Control and Deca) and were exposed to nandrolone decanoate for seven weeks. At the end of the administrations, the animals were euthanized, and the tissues were collected for morphological and biochemical analyses. Data were expressed as mean ± SEM. Tukey and Shapiro-Wilk tests were used. ANOVA and chi-square tests were applied for group comparisons. Differences were considered significant when P<0.05. The results showed an increase in body weight in Wistar rats, while Zebrafish body weight was decreased. In both experiments, the number of atretic follicles increased throughout the in vitro culture, from day 0 to day 7, in the Control group (CTRLr and CTRLz), while in the DECA group (DECAr and DECAz), atretic follicles were reduced from D0 to D7. The antioxidant environment, represented by increased the thiol content, which was significantly higher on day zero in CTRLz compared to CTRLr. SOD activity increased in Zebrafish (group DECAz), while CAT activity decreased in both models (group DECAr and DECAz). In conclusion, the study demonstrated similarity in ovarian physiology between the models exposed or not exposed to nandrolone decanoate, suggesting that, when convenient, researchers could consider changing the experimental model.

Keywords

ovarian follicles, anabolic steroids, cell culture, Danio rerio, Rattus norvegicus

References

Aebi H. Catalase in vitro. Methods Enzymol. 1984;105(C):121-6. http://doi.org/10.1016/S0076-6879(84)05016-3. PMid:6727660.

Alves FL, Oliveira MAF, Morais ANP, Martins SD, Sá NAR, Ceccatto VM, Watanabe YF, Araújo VR. Supraphysiological doses of nandrolone decanoate disrupts spermatogenesis but did not interfere on embryo rate. Naunyn Schmiedebergs Arch Pharmacol. 2023;397:4025-34. http://doi.org/10.1007/s00210-023-02847-1

Andrade GHB, Simão VA, Souza BR, Almeida Chuffa LG, Camargo ICC. Sex steroid receptors profiling is influenced by nandrolone decanoate in the ampulla of the fallopian tube: post-treatment and post-recovery analyses. Tissue Cell. 2018;50:79-88. http://doi.org/10.1016/j.tice.2018.01.001. PMid:29429521.

Araújo VR, Gastal MO, Figueiredo JR, Gastal EL. In vitro culture of bovine preantral follicles: a review. Reprod Biol Endocrinol. 2014;12(1):78. http://doi.org/10.1186/1477-7827-12-78. PMid:25117631.

Araújo VR, Lima-Verde IB, Name KPO, Báo SN, Campello CC, Silva JRV, Rodrigues APR, Figueiredo JR. Bone Morphogenetic Protein-6 (BMP-6) induces atresia in goat primordial follicles cultured in vitro. Pesq Vet Bras. 2010;30(9):770-81. http://doi.org/10.1590/S0100-736X2010000900010.

Bailone RL, Fukushima HCS, Fernandes BHV, Aguiar LK, Corrêa T, Janke H, Setti PG, Roça RO, Borra RC. Zebrafish as an alternative animal model in human and animal vaccination research. Lab Anim Res. 2020;36(1):13. http://doi.org/10.1186/s42826-020-00042-4. PMid:32382525.

Bannister JV, Calabrese L. Assays for superoxide dismutase. Methods Biochem Anal. 1987;32:279-312. http://doi.org/10.1002/9780470110539.ch5. PMid:3033431.

Belardin LB, Simão VA, Leite GAA, Almeida Chuffa LG, Camargo ICC. Dose‐dependent effects and reversibility of the injuries caused by nandrolone decanoate in uterine tissue and fertility of rats. Birth Defects Res B Dev Reprod Toxicol. 2014;101(2):168-77. http://doi.org/10.1002/bdrb.21104. PMid:24753332.

Boff SR. Esteroides anabólicos e exercício: ação e efeitos colaterais. Rev Bras Ciência Mov. 2010;18(1):81-8. http://doi.org/10.18511/0103-1716/rbcm.v18n1p81-88.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72(1-2):248-54. http://doi.org/10.1016/0003-2697(76)90527-3. PMid:942051.

Brasil. Resolução Normativa CONCEA nº 34, de 27 de julho de 2017. Diário Oficial da União; Brasília; 31 julho 2017.

Camargo ICC, Gaspar ALC, Frei F, Mesquita SFP. Efeitos dos esteroides anabólicos androgênicos sobre o útero e parâmetros reprodutivos de ratas adultas. Rev Bras Ginecol Obstet. 2009b;31(9):453-60. http://doi.org/10.1590/S0100-72032009000900006. PMid:19876577.

Camargo ICC, Leite GAA, Pinto T, Ribeiro-Paes JT. Histopathologycal f indings in the ovaries and uterus of albino female rats promoted by co- administration of synthetic steroids and nicotine. Exp Toxicol Pathol. 2014;66(4):195-202. http://doi.org/10.1016/j.etp.2014.01.005. PMid:24556002.

Cannavò S, Curtò L, Trimarchi F. Exercise-related female reproductive dysfunction. J Endocrinol Invest. 2001;24(10):823-32. http://doi.org/10.1007/BF03343935. PMid:11765055.

Clark AS, Fast AS. Comparison of the effects of 17 α-methyltestosterone, methandrostenolone, and nandrolone decanoate on the sexual behavior of castrated male rats. Behav Neurosci. 1996;110(6):1478-86. http://doi.org/10.1037/0735-7044.110.6.1478. PMid:8986348.

Dixon D, Alison R, Bach U, Colman K, Foley GL, Harleman JH, Haworth R, Herbert R, Heuser A, Long G, Mirsky M, Regan K, Van Esch E, Westwood FR, Vidal J, Yoshida M. Nonproliferative and proliferative lesions of the rat and mouse female reproductive system. J Toxicol Pathol. 2014;27(3-4, Suppl):1S-107S. http://doi.org/10.1293/tox.27.1S. PMid:25516636.

Dutra BSC, Pagani MM, Ragnini MP. Anabolic steroids: a theoretical approach. Sci J Fac Educ Environ. 2012;3(2):21-39.

Faltermann S, Hettich T, Küng N, Fent K. Effects of the glucocorticoid clobetasol propionate and its mixture with cortisol and different class steroids in adult female zebrafish. Aquat Toxicol. 2020;218:105372. http://doi.org/10.1016/j.aquatox.2019.105372. PMid:31812088.

Furtado RL, Martins J, Oliveira M, Guerreiro DD, de Sá N, Ferraz A, Ceccatto VM, Rodrigues A, Araújo VR. Acute effect of high-intensity interval training exercise on redox status in the ovaries of rats fed a high-fat diet. Reprod Fertil Dev. 2021;33(12):713-24. http://doi.org/10.1071/RD20326. PMid:34437833.

Gervásio CG, Bernuci MP, Silva-de-Sá MF, Rosa-e-Silva ACJS. The role of androgen hormones in early follicular development. ISRN Obstet Gynecol. 2014;2014:1-11. http://doi.org/10.1155/2014/818010.

Goldman JM, Murr AS, Cooper RL. The rodent estrous cycle: characterization of vaginal cytology and its utility in toxicological studies. Birth Defects Res B Dev Reprod Toxicol. 2007;80(2):84-97. http://doi.org/10.1002/bdrb.20106. PMid:17342777.

Gomes F, Almeida Chuffa LG, Fávaro WJ, Scarano WR, Melo-Neto JS, Pinheiro PFF, Domeniconi RF. Nandrolone decanoate and resistance exercise affect prostate morphology and hormone receptor interface in adult rats with implications for the aging process. Andrology. 2020;8(1):211-20. http://doi.org/10.1111/andr.12626. PMid:31012277.

Grier HJ, Lo Nostro F. The germinal epithelium in fish gonads: the unifying concept. In: Norberg B, Kjesbu O.S., Taranger G.L., Andersson E., Stefansson S.O., editors. Proceedings of the 6th International Symposium on the Reproductive Physiology of Fish. Norway: University of Bergen. p.233-236, 2000.

Harper C, Lawrence C. The laboratory zebrafish. Boca Raton: CRC Press; 2016. http://doi.org/10.1201/b13588.

Hoffman JR, Ratamess NA. Medical issues associated with anabolic steroid use: are they exaggerated? J Sports Sci Med. 2006;5(2):182-93. PMid:24259990.

Hulshof SC, Figueiredo JR, Beckers JF, Bevers MM, Van Den Hurk R. Isolation and characterization of preantral follicles from foetal bovine ovaries. Vet Q. 1994;16(2):78-80. http://doi.org/10.1080/01652176.1994.9694423. PMid:7985360.

Iriart JAB, Chaves JC, Orleans RG. Culto ao corpo e uso de anabolizantes entre praticantes de musculação. Cad Saude Publica. 2009;25(4):773-82. http://doi.org/10.1590/S0102-311X2009000400008. PMid:19347203.

Itohan ORI, Abimbola EO, Ehizokhale ES, Willie DN. Effect of nandrolone decanoate on voice and reproductive tissues in wistar rat: histopathological and morphometric assessment. Malaysian J Med Res. 2020;4(2):1-7. http://doi.org/10.31674/mjmr.2020.v04i02.001.

Karbalay-Doust S, Noorafshan A, Ardekani FM, Mirkhani H, Baker G. The revers-ibility of sperm quality after discontinuing nandrolone decanoate in adult male rats. Asian J Androl. 2007;9(2):235-9. http://doi.org/10.1111/j.1745-7262.2007.00203.x. PMid:16855768.

Lee JA, Renshaw SA. Zebrafish screens for new colitis treatments: a bottom-up approach. FEBS J. 2017;284(3):399-401. http://doi.org/10.1111/febs.14005. PMid:28168859.

Marcondes FK, Bianchi FJ, Tanno AP. Determination of the estrous cycle phases of rats: some helpful considerations. Braz J Biol. 2002;62(4A):609-14. http://doi.org/10.1590/S1519-69842002000400008. PMid:12659010.

Mattaraia VGM, Moura ASAMT. Produtividade de ratos Wistar em diferentes sistemas de acasalamento. Cienc Rural. 2012;42(8):1490-6. http://doi.org/10.1590/S0103-84782012000800026.

Migliaccio M, Chioccarelli T, Ambrosino C, Suglia A, Manfrevola F, Carnevali O, Fasano S, Pierantoni R, Cobellis G. Characterization of follicular atresia responsive to BPA in zebrafish by morphometric analysis of follicular stage progression. Int J Endocrinol. 2018;2018:4298195. http://doi.org/10.1155/2018/4298195. PMid:30675159.

Patanè FG, Liberto A, Maglitto ANM, Malandrino P, Esposito M, Amico F, Cocimano G, Rosi GL, Condorelli D, Di Nunno N, Montana A. Nandrolone decanoate: use, abuse and side effects. Medicina. 2020;56(11):1-24. http://doi.org/10.3390/medicina56110606. PMid:33187340.

Pope HG Jr, Katz DL. Psychiatric and medical effects of anabolic- androgenic steroid use: a controlled study of 160 athletes. Arch Gen Psychiatry. 1997;51(5):375-82. http://doi.org/10.1001/archpsyc.1994.03950050035004. PMid:8179461.

Saddick SY. The impact of nandrolone decanoate administration on ovarian and uterine tissues in rat: luteinizing hormone profile, histopathological and morphometric assessment. Saudi J Biol Sci. 2018;25(3):507-12. http://doi.org/10.1016/j.sjbs.2017.08.015. PMid:29692652.

Selman K, Wallace RA, Sarka A, Qi X. Stages of oocyte development in the zebrafish, Brachydanio rerio. J Morphol. 1993;218(2):203-24. http://doi.org/10.1002/jmor.1052180209. PMid:29865471.

Shahidi NT. A review of chemistry, biological action, and clinical applications of anabolic-androgenic steroids. Clin Ther. 2001;23(9):1355-90. http://doi.org/10.1016/S0149-2918(01)80114-4. PMid:11589254.

Silva AECM, Souza BR, Simão VA, Camargo ICC. A busca pelo corpo perfeito vale a pena? Uma abordagem geral. In Anais do 8º Congresso de Extensão Universitária da UNESP; 2015; Bauru. Bauru: UNESP; 2015. p. 1-6

Silva LA. Maturação e fertilização in vitro de oócitos estádio III de zebrafish [dissertação]. Porto Alegre: Programa de Pós-graduação em Zootecnia, Faculdade de Agronomia, Universidade Federal do Rio Grande do Sul; 2015.

Simão VA, Almeida Chuffa LG, Camargo ICC. Ovarian sex steroid receptors and sex hormones in androgenized rats. Reproduction. 2016;152(5):545-59. http://doi.org/10.1530/REP-16-0233. PMid:27562815.

Simão VA, Berloffa Belardin L, Araújo Leite GA, Almeida Chuffa LG, Camargo ICC. Effects of different doses of nandrolone decanoate on estrous cycle and ovarian tissue of rats after treatment and recovery periods. International Journal of Experimental Pathology © 2015. Int J Exp Pathol. 2015;96(5):338-49. http://doi.org/10.1111/iep.12144.

Simão VA, Lupi LA Jr, Leite GAA, Camargo ICC, Almeida Chuffa LG. Nandrolone decanoate causes uterine injury by changing hormone levels and sex steroid receptors in a dose- and time-dependent manner. Reprod Toxicol. 2021;102:98-108. http://doi.org/10.1016/j.reprotox.2021.05.002. PMid:33984419.

Souza BR, Almeida Chuffa LG, Simão VA, Camargo ICC. Histopathological changes in androgenized ovaries are recovered by melatonin treatment. Int J Exp Pathol. 2018;99(4):158-71. http://doi.org/10.1111/iep.12283. PMid:30256483.

Takahashi H, Nara Y, Tuzimura K. Fluorometric determination of glutathione by N-(9-Acridinyl)-maleimide and its application to biological materials. Agric Biol Chem. 1978;42(4):769-74. http://doi.org/10.1080/00021369.1978.10863059.

Tamaki T, Shiraishi T, Takeda H, Matsumiya T, Roy RR, Edgerton VR. Nandrolone decanoate enhances hypothalamic biogenic amines in rats. Med Sci Sports Exerc. 2003;35(1):32-8. http://doi.org/10.1097/00005768-200301000-00006. PMid:12544632.

Tokarz RR. Oogonial proliferation, oogenesis, and folliculogenesis in nonmammalian vertebrates. In: Jones RE, editors. The vertebrate ovary. New York: Plenum Press; 1978. p. 145-179.

Westerfield M. The zebrafish book: a guide for the laboratory use of zebrafish (Danio rerio). Eugene: University of Oregon Press; 2007.

Zaami S, Tini A, Varì MR. Commentary - Increasing abuse of anabolic steroids and chemsex drugs as performance and image-enhancing agents. Eur Rev Med Pharmacol Sci. 2021;25(1):455-8. PMid:33506936.

Submitted date:
04/14/2024

Accepted date:
11/05/2024

Comparative ovarian morphophysiology of Wistar rats and Zebrafish after exposure to nandrolone decanoate

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