Reproductive Sciences

, Volume 19, Issue 2, pp 163–172 | Cite as

Bisphenol A Induces Oxidative Stress and Decreases Levels of Insulin Receptor Substrate 2 and Glucose Transporter 8 in Rat Testis

  • Shereen C. D’Cruz
  • R. Jubendradass
  • Premendu P. MathurEmail author
Original Articles


Bisphenol A (BPA), a monomer present in plastics, is known to impair male reproductive functions. Testis executes high-energy-demanding processes such as spermatogenesis and steroidogenesis, the successful accomplishment of which requires several factors including glucose. In this context, we sought to investigate the effects of low doses of BPA on glucose metabolism in the testis of rats and to delineate whether oxidative stress has any role to play in mediating the effects. Bisphenol A was orally administered to rats at dose levels of 0.005, 0.5, 50, and 500 µg/kg body weight for 45 days. A positive control was maintained by orally administering 17β-estradiol at a dose of 50 µg/kg body weight. The levels of plasma glucose and insulin were significantly increased, whereas the testicular glucose level significantly decreased following exposure to BPA and estradiol. A dose-dependent increase in the level of hydrogen peroxide (H2O2) and a significant decline in the activities of hexokinase and phosphofructokinase was observed in the testis of rats treated with BPA. Western blot analyses of insulin receptor substrate 2 (IRS-2) and glucose transporter 8 (GLUT-8) in the testis showed a decline in the levels of these proteins following BPA administration. Immunolocalization of GLUT-8 protein in the testis revealed decreased expression of this protein in spermatocytes and developing spermatids of rats exposed to BPA. The results suggest that persistent exposure to low doses of BPA could disturb glucose homeostasis in the testis and thereby impair testicular functions.


bisphenol A testis oxidative stress insulin receptor substrate 2 glucose transporter 8 


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  1. 1.
    Chapin RE, Adams J, Boekelheide K, et al. NTP-CERHR expert panel report on the reproductive and developmental toxicity of bisphenol A. Birth Defects Res B Dev Reprod Toxicol. 2008; 83(3):157–395.Google Scholar
  2. 2.
    Chitra KC, Latchoumycandane C, Mathur PP. Induction of oxidative stress by bisphenol A in the epididymal sperm of rats. Toxicology. 2003;185(1–2):119–127.Google Scholar
  3. 3.
    vom Saal FS, Cooke PS, Buchanan DL, et al. A physiologically based approach to the study of bisphenol A and other estrogenic chemicals on the size of reproductive organs, daily sperm production, and behavior. Toxicol Ind Health. 1998;14(1–2):239–260.Google Scholar
  4. 4.
    Li MW, Mruk DD, Lee WM, Cheng CY. Disruption of the blood-testis barrier integrity by bisphenol A in vitro: is this a suitable model for studying blood-testis barrier dynamics? Int J Biochem Cell Biol. 2009;41(11):2302–2314.Google Scholar
  5. 5.
    Alonso-Magdalena P, Morimoto S, Ripoll C, Fuentes E, Nadal A. The estrogenic effect of bisphenol A disrupts pancreatic beta-cell function in vivo and induces insulin resistance. Environ Health Perspect. 2006;114(1):106–112.Google Scholar
  6. 6.
    Alonso-Magdalena P, Ropero AB, Soriano S, Quesada I, Nadal A. Bisphenol-A: a new diabetogenic factor? Hormones (Athens). 2010;9(2):118–126.Google Scholar
  7. 7.
    Rommerts FF, Cooke BA, Van der Kemp JW, Van der Molen HJ. Effect of luteinizing hormone on 3’,5’-cyclic AMP and testosterone production in isolated interstitial tissue of rat testis. FEBS Lett. 1973;33(1):114–118.Google Scholar
  8. 8.
    Murono EP, Lin T, Osterman J, Nankin HR. Relationship between inhibition of interstitial cell testosterone synthesis by cytochalasin B and glucose. Biochem Biophys Res Commun. 1982;104(1):299–306.Google Scholar
  9. 9.
    Ibberson M, Uldry M, Thorens B. GLUTX1, a novel mammalian glucose transporter expressed in the central nervous system and insulin-sensitive tissues. J Biol Chem. 2000;275(7):4607–4612.Google Scholar
  10. 10.
    Kokk K, Verajankorva E, Laato M, Wu XK, Tapfer H, Pollanen P. Expression of insulin receptor substrates 1–3, glucose transporters GLUT-1-4, signal regulatory protein 1alpha, phosphatidylinositol 3-kinase and protein kinase B at the protein level in the human testis. Anat Sci Int. 2005;80(2):91–96.Google Scholar
  11. 11.
    Cai D, Yuan M, Frantz DF, et al. Local and systemic insulin resistance resulting from hepatic activation of IKK-beta and NF-kappaB. Nat Med. 2005;11(2):183–190.Google Scholar
  12. 12.
    Park SY, Choi GH, Choi HI, Ryu J, Jung CY, Lee W. Depletion of mitochondrial DNA causes impaired glucose utilization and insulin resistance in L6 GLUT4myc myocytes. J Biol Chem. 2005; 280(11):9855–9864.PubMedCrossRefPubMedCentralGoogle Scholar
  13. 13.
    Archuleta TL, Lemieux AM, Saengsirisuwan V, et al. Oxidant stress-induced loss of IRS-1 and IRS-2 proteins in rat skeletal muscle: role of p38 MAPK. Free Radic Biol Med. 2009;47(10): 1486–1493.Google Scholar
  14. 14.
    Ohta Y, Kinugawa S, Matsushima S, et al. Oxidative stress impairs insulin signal in skeletal muscle and causes insulin resistance in post-infarct heart failure. Am J Physiol Heart Circ Physiol. 2011; doi: 10.1152/01185.2009.Google Scholar
  15. 15.
    Bindhumol V, Chitra KC, Mathur PP. Bisphenol A induces reactive oxygen species generation in the liver of male rats. Toxicology. 2003;188(2–3):117–124.Google Scholar
  16. 16.
    Kabuto H, Amakawa M, Shishibori T. Exposure to bisphenol A during embryonic/fetal life and infancy increases oxidative injury and causes underdevelopment of the brain and testis in mice. Life Sci. 2004;74(24):2931–2940.Google Scholar
  17. 17.
    Chitra KC, Mathur PP. Induction of oxidative stress in epididymis of rats after short-term exposure to bisphenol A. Arch Med Res. 2003;1(1):1–16.Google Scholar
  18. 18.
    Sakaue M, Ohsako S, Ishimura R, et al. Bisphenol-A affects spermatogenesis in the adult rat even at a low dose. J Occup Health. 2001;43(4):185–190.Google Scholar
  19. 19.
    vom Saal FS, Cooke PS, Buchanan DL, et al. A physiologically based approach to the study of bisphenol A and other estrogenic chemicals on the size of reproductive organs, daily sperm production, and behavior. Toxicol Ind Health. 1998;14(1–2):239–260.Google Scholar
  20. 20.
    CPCSEA. CPCSEA guidelines for laboratory animal facility. Indian J Pharmacol. 2003;35(4):257–274.Google Scholar
  21. 21.
    EFSA. Opinion of the scientific panel on food additives, flavourings, processing aids and materials in contact with food on a request from the commission related to 2,2-bis(4-hydroxyphenyl) propane (bisphenol A). EFSA J. 2006;428:1–75.Google Scholar
  22. 22.
    Melner MH, Abney TO. The direct effect of 17 beta-estradiol on LH-stimulated testosterone production in hypophysectomized rats. J Steroid Biochem. 1980;13(2):203–210.Google Scholar
  23. 23.
    Gordon MN, Osterburg HH, May PC, Finch CE. Effective oral administration of 17 beta-estradiol to female C57BL/6J mice through the drinking water. Biol Reprod. 1986;35(5):1088–1095.Google Scholar
  24. 24.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265–275.Google Scholar
  25. 25.
    Trinder P. Determination of blood glucose using an oxidase-peroxidase system with a non-carcinogenic chromogen. J Clin Pathol. 1969;22(2):158–161.Google Scholar
  26. 26.
    Pick E, Keisari Y. Superoxide anion and hydrogen peroxide production by chemically elicited peritoneal macrophages—induction by multiple nonphagocytic stimuli. Cell Immunol. 1981; 59(2):301–318.Google Scholar
  27. 27.
    Bergmeyer HU, Grabl M, Walter HE. Methods of Enzymatic Analysis. Deerfield Beach, FL: Verlag Chemie; 1983:222–223.Google Scholar
  28. 28.
    Narabayashi H, Lawson JW, Uyeda K. Regulation of phosphofructokinase in perfused rat heart. Requirement for fructose 2,6-bisphosphate and a covalent modification. J Biol Chem. 1985;260(17):9750–9758.Google Scholar
  29. 29.
    Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259): 680–685.Google Scholar
  30. 30.
    Cranmer M, Louie S, Kennedy RH, Kern PA, Fonseca VA. Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is associated with hyperinsulinemia and insulin resistance. Toxicol Sci. 2000; 56(2):431–436.Google Scholar
  31. 31.
    Turyk M, Anderson H, Knobeloch L, Imm P, Persky V. Organo-chlorine exposure and incidence of diabetes in a cohort of Great Lakes sport fish consumers. Environ Health Perspect. 2009; 117(7):1076–1082.PubMedPubMedCentralCrossRefGoogle Scholar
  32. 32.
    Nadal A, Alonso-Magdalena P, Soriano S, Quesada I, Ropero AB. The pancreatic beta-cell as a target of estrogens and xenoestrogens: Implications for blood glucose homeostasis and diabetes. Mol Cell Endocrinol. 2009;304(1–2):63–68.Google Scholar
  33. 33.
    Godsland IF. Oestrogens and insulin secretion. Diabetologia. 2005;48(11):2213–2220.Google Scholar
  34. 34.
    Kokk K, Verajankorva E, Wu XK, et al. Expression of insulin signaling transmitters and glucose transporters at the protein level in the rat testis. Ann N Y Acad Sci. 2007;1095:262–273.Google Scholar
  35. 35.
    Adachi T, Yasuda K, Mori C, et al. Promoting insulin secretion in pancreatic islets by means of bisphenol A and nonylphenol via intracellular estrogen receptors. Food Chem Toxicol. 2005; 43(5):713–719.Google Scholar
  36. 36.
    Wolff SP, Bascal ZA, Hunt JV. “Autoxidative glycosylation”: free radicals and glycation theory. Prog Clin Biol Res. 1989; 304:259–275.Google Scholar
  37. 37.
    Nishikawa T, Edelstein D, Du XL, et al. Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature. 2000;404(6779):787–790.Google Scholar
  38. 38.
    Kaneto H, Fujii J, Suzuki K, et al. DNA cleavage induced by glycation of Cu, Zn-superoxide dismutase. Biochem J. 1994;304(Pt 1): 219–225.Google Scholar
  39. 39.
    Shin AH, Oh CJ, Park JW. Glycation-induced inactivation of antioxidant enzymes and modulation of cellular redox status in lens cells. Arch Pharm Res. 2006;29(7):577–581.Google Scholar
  40. 40.
    Nakamura M, Okinaga S, Arai K. Metabolism of pachytene primary spermatocytes from rat testes: pyruvate maintenance of adenosine triphosphate level. Biol Reprod. 1984;30(5):1187–1197.Google Scholar
  41. 41.
    Colussi C, Albertini MC, Coppola S, Rovidati S, Galli F, Ghibelli L. H2O2-induced block of glycolysis as an active ADP-ribosylation reaction protecting cells from apoptosis. FASEB J. 2000;14(14):2266–2276.Google Scholar
  42. 42.
    Hegde KR, Kovtun S, Varma SD. Inhibition of glycolysis in the retina by oxidative stress: prevention by pyruvate. Mol Cell Biochem. 2010;343(1–2):101–105.Google Scholar
  43. 43.
    Janero DR, Hreniuk D, Sharif HM. Hydroperoxide-induced oxidative stress impairs heart muscle cell carbohydrate metabolism. Am J Physiol. 1994;266(1 Pt 1):C179–188.Google Scholar
  44. 44.
    Huang X, Vaag A, Hansson M, Groop L. Down-regulation of insulin receptor substrates (IRS)-1 and IRS-2 and Src homologous and collagen-like protein Shc gene expression by insulin in skeletal muscle is not associated with insulin resistance or type 2 diabetes. J Clin Endocrinol Metab. 2002;87(1):255–259.Google Scholar
  45. 45.
    Zhang J, Ou J, Bashmakov Y, Horton JD, Brown MS, Goldstein JL. Insulin inhibits transcription of IRS-2 gene in rat liver through an insulin response element (IRE) that resembles IREs of other insulin-repressed genes. Proc Natl Acad Sci U S A. 2001;98(7): 3756–3761.PubMedPubMedCentralCrossRefGoogle Scholar
  46. 46.
    Ibberson M, Riederer BM, Uldry M, Guhl B, Roth J, Thorens B. Immunolocalization of GLUTX1 in the testis and to specific brain areas and vasopressin-containing neurons. Endocrinology. 2002; 143(1):276–284.Google Scholar
  47. 47.
    Doege H, Schurmann A, Bahrenberg G, Brauers A, Joost HG. GLUT8, a novel member of the sugar transport facilitator family with glucose transport activity. J Biol Chem. 2000;275(21): 16275–16280.Google Scholar

Copyright information

© Society for Reproductive Investigation 2012

Authors and Affiliations

  • Shereen C. D’Cruz
    • 1
  • R. Jubendradass
    • 1
  • Premendu P. Mathur
    • 1
    Email author
  1. 1.Department of Biochemistry and Molecular Biology, School of Life SciencesPondicherry UniversityPondicherryIndia

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