Journal of Toxicology and Environmental Health Sciences
Subscribe to JTEHS
Full Name*
Email Address*

Article Number - 111C3E847679


Vol.6(8), pp. 161-169 , October 2014
DOI: 10.5897/JTEHS2014.0314
ISSN: 2006-9820



Full Length Research Paper

Effect of dipentyl phthalate in 3-dimensional in vitro testis co-culture is attenuated by cyclooxygenase-2 inhibition



Susanna Wegner
  • Susanna Wegner
  • Institute for Risk Analysis and Risk Communication, School of Public Health, University of Washington, Washington DC, USA
  • Google Scholar
Xiaozhong Yu*
  • Xiaozhong Yu*
  • Institute for Risk Analysis and Risk Communication, School of Public Health, University of Washington, Washington DC, USA
  • Google Scholar
Hee Yeon Kim
  • Hee Yeon Kim
  • Institute for Risk Analysis and Risk Communication, School of Public Health, University of Washington, Washington DC, USA
  • Google Scholar
Sean Harris
  • Sean Harris
  • Institute for Risk Analysis and Risk Communication, School of Public Health, University of Washington, Washington DC, USA
  • Google Scholar
William C. Griffith
  • William C. Griffith
  • Institute for Risk Analysis and Risk Communication, School of Public Health, University of Washington, Washington DC, USA
  • Google Scholar
Sungwoo Hong
  • Sungwoo Hong
  • Institute for Risk Analysis and Risk Communication, School of Public Health, University of Washington, Washington DC, USA
  • Google Scholar
Elaine M. Faustman
  • Elaine M. Faustman
  • Institute for Risk Analysis and Risk Communication, School of Public Health, University of Washington, Washington DC, USA
  • Google Scholar







 Received: 16 August 2014  Accepted: 15 September 2014  Published: 31 October 2014

Copyright © 2014 Author(s) retain the copyright of this article.
This article is published under the terms of the Creative Commons Attribution License 4.0


Exposure to phthalate esters is associated with changes in steroidogenesis, leading to the hypothesis that this is a primary mechanism of phthalate reproductive toxicity. However, some phthalate-induced male reproductive toxicity has been demonstrated in the absence of changes to testosterone production, suggesting additional mechanisms of action. There is evidence that phthalate exposure increases expression of the inflammatory enzyme cyclooxygenase 2 (cox-2). Furthermore, inhibition of cox-2 enhances expression of the steroidogenic acute regulatory protein (StAR), which mediates the rate-limiting step in steroidogenesis. This study hypothesized that phthalate-induced toxicity and testosterone perturbation are mediated in part by cox-2. A 3D in vitro rat testis co-culture to explore the role of cox-2 in phthalate toxicity was employed. Cells were treated with 100 µM dipentyl phthalate (DPP) with and without pre-treatment with the specific cox-2 inhibitor NS-398. Effects were evaluated after 8, 24, and 72 h. DPP exposure significantly increased cox-2 expression at 8 and 24 h (p<0.01) and resulted in significant, dose-dependent cytotoxicity. Pre-treatment with NS-398 significantly reduced the cytotoxicity of DPP at 8 and 24 h (p<0.01). NS-398 also mitigated the effects of DPP on testosterone regulation. Total testosterone concentrations in cell culture media were significantly increased following 8 and 24 hr of DPP exposure (p<0.001) and NS-398 reduced this effect (p<0.05). Simultaneously, DPP significantly decreased StAR protein expression after 8 h (p<0.01) and this effect was significantly attenuated by the presence of NS-398 (p<0.01). These results suggest that the DPP-induced changes in testosterone regulation observed in this experiment are mediated in part by an inflammatory response that is cox-2 dependent.

 

Key words: dipentyl phthalate, testosterone, cyclooxygenase 2, in vitro toxicology        

Alam MS, Ohsako S, Matsuwaki T, Zhu XB, Tsunekawa N, Kanai Y, Sone H, Tohyama C, Kurohmaru M (2010). Induction of spermatogenic cell apoptosis in prepubertal rat testes irrespective of testicular steroidogenesis: a possible estrogenic effect of di(n-butyl) phthalate. Reproduction 139 (2):427-37.
Crossref
 
Barlow NJ, Phillips SL, Wallace DG, Sar M, Gaido KW, Foster PM (2003). Quantitative changes in gene expression in fetal rat testes following exposure to di(n-butyl) phthalate.Toxicol.Sci.73(2):431-41.
Crossref
 
Bornehag CG, Nanberg E (2010). Phthalate exposure and asthma in children. Int. J. Androl. 33(2):333-45.
Crossref
 
Clark BJ, Cochrum RK (2007). The steroidogenic acute regulatory protein as a target of endocrine disruption in male reproduction. Drug Metab. Rev. 39 (2-3):353-70.
Crossref
 
David RM (2006). Proposed mode of action for in utero effects of some phthalate esters on the developing male reproductive tract. Toxicol. Pathol. 34(3):209-19.
Crossref
 
Ferguson KK, Loch-Caruso R, Meeker JD (2011). Urinary phthalate metabolites in relation to biomarkers of inflammation and oxidative stress: NHANES 1999-2006. Environ. Res. 111(5):718-26.
Crossref
 
Foster PM, Thomas LV, Cook MW, Gangolli SD (1980). Study of the testicular effects and changes in zinc excretion produced by some n-alkyl phthalates in the rat. Toxicol. Appl. Pharmacol. 54(3):392-8. Fromme H, Gruber L, Seckin E, Raab U, Zimmermann S, Kiranoglu M, Schlummer M, Schwegler U, Smolic S, Volkel W (2011). Phthalates and their metabolites in breast milk--results from the Bavarian Monitoring of Breast Milk (BAMBI). Environ. Int. 37(4):715-22.
 
Gaido KW, Hensley JB, Liu D, Wallace DG, Borghoff S, Johnson KJ, S Hall SJ, Boekelheide K (2007). Fetal mouse phthalate exposure shows that Gonocyte multinucleation is not associated with decreased testicular testosterone. Toxicol. Sci. 97 (2):491-503.
Crossref
 

Granholm T, Creasy DM, Pollanen P, Soder O (1992). Di-n-pentyl phthalate-induced inflammatory changes in the rat testis are accom-panied by local production of a novel lymphocyte activating factor. J Reprod. Immunol. 21(1):1-14.

Gray LE Jr, Ostby J, FurrJ, Price M, Veeramachaneni DN, Parks L (2000). Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat. Toxicol. Sci. 58(2):350-65.

 
Hannas BR, Furr J, Lambright CS, Wilson VS, Foster PM, Gray LE Jr (2011). Dipentyl phthalate dosing during sexual differentiation disrupts fetal testis function and postnatal development of the male Sprague-Dawley rat with greater relative potency than other phthalates. Toxicol. Sci. 120(1):184-93.
Crossref
 
Hauser R, Calafat AM (2005). Phthalates and human health. Occup. Environ. Med. 62(11):806-18.
Crossref
 
Heger NE, Hall SJ, Sandrof MA, McDonnell EV, Hensley JB, McDowell EN, Martin KA, Gaido KW, Johnson KJ Boekelheide K (2012). Human fetal testis xenografts are resistant to phthalate-induced endocrine disruption. Environ. Health Perspect. 120(8):1137-43.
Crossref
 
Howdeshell KL, Rider CV, Wilson VS, Gray LE Jr (2008). Mecha-nisms of action of phthalate esters, individually and in combination, to induce abnormal reproductive development in male laboratory rats. Environ. Res. 108(2):168-76.
Crossref
 
Ishikawa T, Hwang K, Lazzarino D, Morris PL (2005). Sertoli cell expression of steroidogenic acute regulatory protein-related lipid transfer 1 and 5 domain-containing proteins and sterol regulatory element binding protein-1 are interleukin-1beta regulated by activation of c-Jun N-terminal kinase and cyclooxygenase-2 and cytokine induction. Endocrinology 146(12):5100-11.
Crossref
 
Johnson KJ, Heger NE, Boekelheide. K (2012). Of mice and men (and rats): phthalate-induced fetal testis endocrine disruption is species-dependent. Toxicol. Sci. 129(2):235-48.
Crossref
 
Jurewicz J, Radwan M, Sobala W, Ligocka D, Radwan P, Bochenek M, Hawula W, Jakubowski L, Hanke W (2013). Human urinary phthalate metabolites level and main semen parameters, sperm chromatin structure, sperm aneuploidy and reproductive hormones. Reprod. Toxicol. 42:232-41.
Crossref
 
Kimber I, Dearman RJ (2010). An assessment of the ability of phthalates to influence immune and allergic responses. Toxicology 271(3):73-82.
Crossref
 
Kristensen DM, Skalkam ML, Audouze K, Lesne L, Desdoits-Lethimonier C, Frederiksen H, Brunak S, Skakkebaek NE, Jegou B, Hansen JB, Junker S, Leffers H (2011). Many putative endocrine disruptors inhibit prostaglandin synthesis. Environ. Health Perspect. 119(4):534-41.
Crossref
 
Lahousse SA, Wallace DG, Liu D, Gaido KW, Johnson KJ (2006). Testicular gene expression profiling following prepubertal rat mono-(2-ethylhexyl) phthalate exposure suggests a common initial genetic response at fetal and prepubertal ages. Toxicol. Sci. 93(2):369-81.
Crossref
 
Lambrot R, Muczynski V, Lecureuil C, Angenard G, Coffigny H, Pairault C, Moison D, Frydman R, Habert R, Rouiller-Fabre V (2009). Phthalates impair germ cell development in the human fetal testis in vitro without change in testosterone production. Environ. Health Perspect. 117 (1):32-7.
Crossref
 
Lehmann KP, Phillips S, Sar M, Foster PM, Gaido KW (2004). Dose-de-pendent alterations in gene expression and testosterone synthesis in the fetal testes of male rats exposed to di (n-butyl) phthalate. Toxicol. Sci. 81(1):60-8.
Crossref
 
Murphy CJ, Stermer AR, Richburg.JH (2014). Age- and Species-Dependent Infiltration of Macrophages into the Testis of Rats and Mice Exposed to Mono-(2-Ethylhexyl) Phthalate (MEHP). Biol. Reprod. biolreprod-113.
 
Mylchreest E, Cattley RC, Foster PM (1998). Male reproductive tract malformations in rats following gestational and lactational exposure to Di(n-butyl) phthalate: an antiandrogenic mechanism? Toxicol. Sci. 43(1):47-60.
Crossref
 
Onorato TM, Brown PW, Morris PL (2008). Mono-(2-ethylhexyl) phthalate increases spermatocyte mitochondrial peroxiredoxin 3 and cyclooxygenase 2. J. Androl. 29(3):293-303.
Crossref
 
Rock KL, Kono H (2008). The inflammatory response to cell death. Annu. Rev. Pathol. 3:99-126.
Crossref
 
Silva MJ, Reidy JA, Herbert AR, Preau JL Jr, Needham LL, Calafat AM (2004). Detection of phthalate metabolites in human amniotic fluid. Bull. Environ. Contam. Toxicol. 72(6):1226-31.
Crossref
 
Swan SH (2008). Environmental phthalate exposure in relation to repro-ductive outcomes and other health endpoints in humans. Environ. Res. 108(2):177-84.
Crossref
 
Swan SH, Liu F, Hines M, Kruse RL, Wang C, Redmon JB, Sparks A, Weiss B (2010). Prenatal phthalate exposure and reduced masculine play in boys. Int. J. Androl. 33(2):259-69.
Crossref
 
Swan SH, Main KM, Liu F, Stewart SL, Kruse RL, Calafat AM, Mao CS, Redmon JB, Ternand CL, Sullivan S. Teague. JL (2005). Decrease in anogenital distance among male infants with prenatal phthalate exposure. Environ. Health Perspect. 113(8):1056-61.
Crossref
 
Wang X, Dyson MT, Jo Y, Stocco DM (2003). Inhibition of cyclooxygenase-2 activity enhances steroidogenesis and steroidogenic acute regulatory gene expression in MA-10 mouse Leydig cells. Endocrinol. 144(8):3368-75.
Crossref
 
Wang X, Shen CL, Dyson MT, Eimerl S, J. Orly J, Hutson JC, D. M. Stocco DM (2005). Cyclooxygenase-2 regulation of the age-related decline in testosterone biosynthesis. Endocrinology 146(10):4202-8.
Crossref
 
Wegner S, Hong S, Yu X, Faustman EM (2013). Preparation of rodent testis co-cultures. Curr. Protoc. Toxicol. 16-10.
 
Yu X, Hong S, Moreira EG, Faustman EM (2009). Improving in vitro Sertoli cell/gonocyte co-culture model for assessing male reproduc-tive toxicity: Lessons learned from comparisons of cytotoxicity versus genomic responses to phthalates. Toxicol. Appl. Pharmacol. 239(3):325-36.
Crossref
 
Yu X, Sidhu JS, Hong S, Faustman. EM (2005). Essential role of extra-cellular matrix (ECM) overlay in establishing the functional integrity of primary neonatal rat Sertoli cell/gonocyte co-cultures: an improved in vitro model for assessment of male reproductive toxicity. Toxicol. Sci. 84(2):378-93.
Crossref

 


APA Wegner, S., Yu, X., Kim, Y.H., Harris, S., Griffith, W.C., Hong, S. & Faustman, E.M. (2014). Effect of dipentyl phthalate in 3-dimensional in vitro testis co-culture is attenuated by cyclooxygenase-2 inhibition. Journal of Toxicology and Environmental Health Sciences, 6(8), 161-169.
Chicago Susanna Wegner, Xiaozhong Yu, Hee Yeon Kim, Sean Harris, William C. Griffith, Sungwoo Hong and Elaine M. Faustman. "Effect of dipentyl phthalate in 3-dimensional in vitro testis co-culture is attenuated by cyclooxygenase-2 inhibition." Journal of Toxicology and Environmental Health Sciences 6, no. 8 (2014): 161-169.
MLA Susanna Wegner, et al. "Effect of dipentyl phthalate in 3-dimensional in vitro testis co-culture is attenuated by cyclooxygenase-2 inhibition." Journal of Toxicology and Environmental Health Sciences 6.8 (2014): 161-169.
   
DOI 10.5897/JTEHS2014.0314
URL http://academicjournals.org/journal/JTEHS/article-abstract/111C3E847679

Subscription Form