Oxidative Stress and DNA methylation in male rat pups provoked by the transplacental and translactational exposure to bisphenol A

  • Hanan M. A. El Henafy
  • Marwa A. IbrahimEmail author
  • Samy A. Abd El Aziz
  • Eman M. Gouda
Short Research and Discussion Article


The epigenetic changes induced by environmental contaminants play important roles in the inheritance of male reproductive dysfunction. The present study investigated DNA methylation changes and some oxidative stress biomarkers induced by bisphenol A (BPA) in male offspring. A total number of 48 female albino rats were administered orally with 50 μg/kg of BPA/day during gestation and/or lactation periods. At postnatal day 60, the samples were collected from the male pups to assess the serum testosterone, malondialdehyde (MDA) level, superoxide dismutase (SOD), glutathione S-transferase, and glutathione peroxidase (GSH-Px) activities in testicular tissue. DNA methylation in both DNA (cytosine-5)-methyltransferase 3A and estrogen receptor alpha genes was detected by methylation-specific PCR. BPA exposure resulted in significant decrease in the anogenital distance, testis and epididymis weights, serum testosterone level, SOD, GST, and GSH-Px levels with significant increase in weaning body weight and the MDA level. Additionally, BPA caused marked hypermethylation within Dnmt3A and ER- ∝ genes promoter regions in the testis of rat male pups.

Graphical abstract


Bisphenol A Epigenetics DNA hypermethylation Dnmt3A gene ER- ∝ gene 


Author contributions

Samy Abd El Aziz contributed to design the experiment and interpretation of the results. Eman Gouda contributed to design, acquisition, and the analysis of data. Hanan El Henafy contributed to the analysis of data and drafted the manuscript. Marwa Ibrahim contributed to the analysis of data and drafted the manuscript. All authors approved the manuscript and ensured originality and accuracy.

Compliance with ethical standards

Animals were kept and treated according to the guidelines of the ethics committee of Cairo University (approval number CU II S 12 16).

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Abdel Aziz RL, Abdel-Wahab A, Abo El-Ela FI, Hassan NEY, El-Nahass ES, Ibrahim MA, Khalil A (2018) Dose-dependent ameliorative effects of quercetin and l-Carnitine against atrazine-induced reproductive toxicity in adult male Albino rats. Biomed Pharmacother 102:855–864CrossRefGoogle Scholar
  2. Ahbab MA, Barlas N, Karabulut G (2017) The toxicological effects of bisphenol A and octylphenol on the reproductive system of prepubertal male rats. Toxicol Ind Health 33(2):133–146CrossRefGoogle Scholar
  3. Ayyed HH, Abdulrazzak NK, Abdul Ameer AI, Baccarelli A, Bollati V (2009) Epigenetics and environmental chemicals. Curr Opin Pediatr 21:243–251CrossRefGoogle Scholar
  4. Barodia SK, Creed RB, Goldberg MS (2017) Parkin and PINK1 functions in oxidative stress and neurodegeneration. Brain Res Bull 133:51–59CrossRefGoogle Scholar
  5. Chen Z, Zuo X, He D, Ding S, Xu F, Yang H, Jin X, Fan Y, Ying L, Tian C, Ying C (2017) Sci Rep 7:40337CrossRefGoogle Scholar
  6. Cheng YT, Yang CC, Shyur LF (2016) Phytomedicine-Modulating oxidative stress and the tumor microenvironment for cancer therapy. Pharmacol Res 114:128–143CrossRefGoogle Scholar
  7. Chikara S, Nagaprashantha LD, Singhal J, Horne D, Awasthi S, Singhal SS (2018) Oxidative stress and dietary phytochemicals: role in cancer chemoprevention and treatment. Cancer Lett 413:122–134CrossRefGoogle Scholar
  8. Christiansen S, Axelstad M, Boberg J, Vinggaard AM, Pedersen GA, Hass U (2014) Low-dose effects of bisphenol A on early sexual development in male and female rats. Reproduction 147(4):477–487CrossRefGoogle Scholar
  9. de Freitas MA, da Silva AM, Rios AF, Renzi A, Lôbo RB, Galerani MA, Vila RA, Ramos ES (2011) Identification of a DNA methylation point in the promoter region of the bovine CYP21 gene. Genet Mol Res 10(3):1409–1415CrossRefGoogle Scholar
  10. Donkena, Krishna Vanaja; Young, Charles Y. F.; Tindall, Donald J, (2010). Oxidative stress and DNA methylation in prostate cancer. Obstet Gynecol Int. 2010, ID 302051, 14 pages.Google Scholar
  11. Gao L, Wang HN, Zhang L, Peng FY, Jia Y, Wei W, Jia LH (2016) Effect of perinatal bisphenol A exposure on serum lipids and lipid enzymes in offspring rats of different sex. Biomed Environ Sci 29(9):686–689Google Scholar
  12. Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases: the first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139Google Scholar
  13. Hasanein P, Kazemian-Mahtaj A, Khodadadi I (2016) Bioactive peptide carnosin protects against lead acetate-induced hepatotoxicity by abrogation of oxidative stress in rats. Pharm Biol 54(8):1458–1464CrossRefGoogle Scholar
  14. Jain D, Meydan C, Lange J, Claeys Bouuaert C, Lailler N, Mason CE, Anderson KV, Keeney S (2017) Rahu is a mutant allele of Dnmt3c, encoding a DNA methyltransferase homolog required for meiosis and transposon repression in the mouse male germline. PLoS Genet 13:e1006964CrossRefGoogle Scholar
  15. Jin J, Lian T, Gu C, Yu K, Gao YQ, Su XD (2016) The effects of cytosine methylation on general transcription factors. Sci Rep 6:29119CrossRefGoogle Scholar
  16. Johnson SA, Spollen WG, Manshack LK, Bivens NJ, Givan SA, Rosenfeld CS. Hypothalamic transcriptomic alterations in male and female California mice (Peromyscus californicus) developmentally exposed to bisphenol A or ethinyl estradiol. Phys Rep. 2017 ;5(3). pii: e13133.Google Scholar
  17. Kamel S, Ibrahim MA, Awad ET, El-Hindi HMA, Abdel-Aziz SA (2018) Molecular cloning and characterization of the novel CYP2J2 in dromedary camels (Camelus dromedarius). Int J Biol Macromol 120(Pt B):1770–1776CrossRefGoogle Scholar
  18. Kazemi S, Mousavi SN, Aghapour F, Rezaee B, Sadeghi F, Moghadamnia AA (2016) Induction effect of bisphenol A on gene expression involving hepatic oxidative stress in rat. Oxidative Med Cell Longev 6298515Google Scholar
  19. Khairy HM, Ibrahim MA, Ibrahem MD (2012) Phenological and liver antioxidant profiles of adult Nile tilapia (Oreochromis niloticus) exposed to toxic live cyanobacterium (Microcystis aeruginosa Kützing) cells. Z Naturforsch C 67(11-12):620–628CrossRefGoogle Scholar
  20. Khalaf AA, Ibrahim AM, Tohamy AF, Allah AAA, Zaki AR (2017) Protective effect of vitazinc on chlorsan induced oxidatives, genotoxicity and histopathological changes in testicular tissues of male rats. Int J Pharmacol 13:22–32CrossRefGoogle Scholar
  21. Khalaf AA, Ahmed W, Moselhy WA, Abdel-Halim BR, Ibrahim MA (2019) Protective effects of selenium and nano-selenium on bisphenol-induced reproductive toxicity in male rats. Hum Exp Toxicol 38(4):398–408CrossRefGoogle Scholar
  22. Kochmanski J, Montrose L Goodrich JM, Dolinoy DC Environmental deflection: the impact of toxicant exposures on the aging epigenome. Toxicol Sci 2017;156(2):325-335.Google Scholar
  23. Kundakovic M, Kathryn G, Becca F, Jesus M, Rachel LM, Frederica P, Frances AC (2013) Sex-specific epigenetic disruption and behavioral changes following low-dose in utero bisphenol A exposure. Proc Natl Acad Sci U S A 110(24):9956–9961CrossRefGoogle Scholar
  24. Laing LV, Viana J, Dempster EL, Trznadel M, Trunkfield LA, Uren Webster TM, van Aerle R, Paull GC, Wilson RJ, Mill J, Santos EM (2016) Bisphenol A causes reproductive toxicity, decreases dnmt1 transcription, and reduces global DNA methylation in breeding zebrafish (Danio rerio). Epigenetics. 11(7):526–538CrossRefGoogle Scholar
  25. Lama S, Vanacore D, Diano N, Nicolucci C, Errico S, Dallio M, Federico A, Loguercio C, Stiuso P (2019) Ameliorative effect of Silybin on bisphenol A induced oxidative stress, cell proliferation and steroid hormones oxidation in HepG2 cell cultures Scientific Reports. 9(1):3228Google Scholar
  26. Lawrence RA, Burk RF (1976) Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun 71:952–958CrossRefGoogle Scholar
  27. Manikkam M, Guerrero-Bosagna C, Tracey R, Haque MM, Skinner MK (2012) Transgenerational actions of environmental compounds on reproductive disease and identification of epigenetic biomarkers of ancestral exposures. PLoS ONE 7(2):e31901 Long-term exposure to a ‘safe’ dose of bisphenol A reduced proteinacetylation in adult rat testesCrossRefGoogle Scholar
  28. Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474CrossRefGoogle Scholar
  29. Moreman J, Lee O, Trznadel M, David A, Kudoh T, Tyler CR (2017) Acute toxicity, teratogenic, and estrogenic effects of bisphenol A and its alternative replacements bisphenol S, bisphenol F, and bisphenol AF in zebrafish embryo-larvae. Environ Sci Technol 51(21):12796–12805CrossRefGoogle Scholar
  30. Morgan AM, Ibrahim MA, Hussien AM (2017) The potential protective role of Akropower against Atrazine-induced humoral immunotoxicity in rabbits. Biomed Pharmacother 96:710–715CrossRefGoogle Scholar
  31. Morgan A, Ibrahim MA, Galal MK, Ogaly HA, Abd-Elsalam RM (2018) Innovative perception onusing Tiron to modulate the hepatotoxicity induced by titanium dioxide nanoparticles in male rats. Biomed Pharmacother 103:553–561CrossRefGoogle Scholar
  32. Moselhy WA, Helmy NA, Abdel-Halim BR, Nabil TM, Abdel-Hamid MI (2012) Role of ginger against the reproductive toxicity of aluminium chloride in albino male rats. Reprod Domest Anim 47(2):335–343CrossRefGoogle Scholar
  33. Nadal A, Fuentes E, Ripoll C, Villar-Pazos S, Castellano-Muñoz M, Soriano S, Martinez-Pinna J, Quesada I, Alonso-Magdalena P (2018) Extranuclear-initiated estrogenic actions of endocrine disrupting chemicals: is there toxicology beyond paracelsus? J Steroid Biochem Mol Biol 176:16–22CrossRefGoogle Scholar
  34. Nikaido Y, Yoshizawa K, Danbara N, Tsujita-Kyutoku M, Yuri T, Uehara N (2004) Effects of maternal xenoestrogen exposure on development of the reproductive tract and mammary gland in female CD-1 mouse offspring. Reprod Toxicol 18(6):803–811CrossRefGoogle Scholar
  35. Okazaki H, Takeda S, Kakizoe K, Taniguchi A, Tokuyasu M, Himeno T, Ishii H, Kohro-Ikeda E, Haraguchi K, Watanabe K (2017) Bisphenol AF as an inducer of estrogen receptor β (ERβ): evidence for anti-estrogenic effects at higher concentrations in human breast cancer cells. Biol Pharm Bull 40(11):1909–1916CrossRefGoogle Scholar
  36. Pisolato R, Lombardi AP, Vicente CM, Lucas TF, Lazari MF, Porto CS (2016) Expression and regulation of the estrogen receptors in PC-3 human prostate cancer cells. Steroids. 107:74–86CrossRefGoogle Scholar
  37. Pivnenko K, Pedersen GA, Eriksson E, Astrup TF (2015) Bisphenol A and its structural analogues in household waste paper. Waste Manag 44:39–47CrossRefGoogle Scholar
  38. Rashad MM, Galal MK, El-Behairy AM, Gouda EM, Moussa SZ (2018) Maternal exposure to di-n-butyl phthalate induces alterations of c-Myc gene, some apoptotic and growth related genes in pups’ testes. Toxicol Ind Health 34(11):744–752CrossRefGoogle Scholar
  39. Ruiz-Larrea MB, Leal AM, Liza M, Lacort M, de Groot H (1994) Antioxidant effects of estradiol and 2-hydroxyestradiol on iron induced lipid peroxidation of rat liver microsomes. Steroids. 59:383–388CrossRefGoogle Scholar
  40. Santangeli S, Maradonna F, Olivotto I, Piccinetti CC, Gioacchini G, Carnevali O (2017) Gen Comp Endocrinol 245:122–126CrossRefGoogle Scholar
  41. Singh S, Shoei-Lung LS (2012) Epigenetic effects of environmental chemicals bisphenol A and phthalates. Int J Mol Sci 13(8):10143–10153CrossRefGoogle Scholar
  42. Tsikas D, Rothmann S, Schneider JY, Suchy MT, Trettin A, Modun D, Stuke N, Maassen N, Frölich JC (2016 Apr 15) Development, validation and biomedical applications of stable-isotope dilution GC-MS and GC-MS/MS techniques for circulating malondialdehyde (MDA) after pentafluorobenzyl bromide derivatization: MDA as a biomarker of oxidative stress and its relation to 15(S)-8-iso-prostaglandin F2α and nitric oxide (NO). J Chromatogr B Anal Technol Biomed Life Sci 1019:95–111CrossRefGoogle Scholar
  43. Ullah H, Jahan S, Ain QU, Shaheen G, Ahsan N (2016) Effect of bisphenol S exposure on male reproductive system of rats: a histological and biochemical study. Chemosphere. 152:383–391CrossRefGoogle Scholar
  44. Wang YQ, Li YW, Chen QL, Liu ZH (2019) Long-term exposure of xenoestrogens with environmental relevant concentrations disrupted spermatogenesis of zebrafish through altering sex hormone balance, stimulating germ cell proliferation, meiosis and enhancing apoptosis. Environ Pollut 244:486–494CrossRefGoogle Scholar
  45. Zhao F, Li K, Zhao L, Liu J, Suo Q, Zhao J, Wang H, Zhao S (2014) Effect of Nrf2 on rat ovarian tissues against atrazine-induced anti-oxidative response. Int J Clin Exp Pathol 7(6):2780–2789Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Medical Laboratory Department, Faculty of Applied Medical SciencesOctober 6 UniversityCairoEgypt
  2. 2.Biochemistry and Chemistry of Nutrition Department, Faculty of Veterinary MedicineCairo UniversityGizaEgypt

Personalised recommendations