Effects of the environmental endocrine disruptors di-2-ethylhexyl phthalate and mono-2-ethylhexyl phthalate on human sperm function in vitro
Xinyi Sun A B , Wenqiong Chen A , Shiqi Weng A , Tingting Pan A , Xiaonian Hu A , Fang Wang A , Tianxinyu Xia A B , Houyang Chen D and Tao Luo
A C E
+ Author Affiliations
- Author Affiliations
A Institute of Life Science and School of Life Science, Nanchang University, 999 Xuefu Road, Honggutan New District, Nanchang, Jiangxi 330031, China.
B Nanchang University Queen Mary School, Jiangxi Medical College of Nanchang University, Nanchang University, 999 Xuefu Road, Honggutan New District, Nanchang, Jiangxi 330031, China.
C Key Laboratory of Reproductive Physiology and Pathology in Jiangxi Province, 999 Xuefu Road, Honggutan New District, Nanchang, Jiangxi 330031, China.
D Reproductive Medical Center, Jiangxi Maternal and Child Health Hospital, 318/81 Avenue, Nanchang, Jiangxi 330006, China.
E Corresponding author. Email: luotao@ncu.edu.cn
Reproduction, Fertility and Development 32(6) 629-636 https://doi.org/10.1071/RD19164
Submitted: 1 May 2019 Accepted: 20 September 2019 Published: 7 February 2020
Abstract
Di-2-ethylhexyl phthalate (DEHP), a plastic-derived, endocrine-disrupting chemical, has been shown to exhibit male reproductive toxicity. However, its effects on human mature spermatozoa are largely unknown. In this study we investigated the in vitro effects of DEHP and mono-2-ethylhexyl phthalate (MEHP; the main metabolite of DEHP) on sperm function and the mechanisms involved. Human spermatozoa were exposed to phthalates in vitro at the doses that cover the concentrations detected in human semen: 20 nM–8 μM DEHP, 1 nM–20 μM MEHP or a mixture of 20 nM–8 μM DEHP and 1 nM–20 μM MEHP. DEHP and MEHP, alone or in combination, had no effect on the viability, membrane integrity, motility, homeostasis of reactive oxygen species or mitochondrial activity of human spermatozoa. Interestingly, 1 nM–20 μM MEHP and combinations of 20 nM–8 μM DEHP and 1 nM–20 μM MEHP enhanced penetration ability, hyperactivation and the spontaneous acrosome reaction of human spermatozoa, and increased intracellular free Ca2+ concentrations ([Ca2+]i) and tyrosine phosphorylation, two key signalling pathways that regulate sperm function. The findings of this study suggest that in vitro exposure to MEHP metabolised from DEHP affects human sperm function by inducing increases in sperm [Ca2+]i and tyrosine phosphorylation, which adds to our understanding of the effects of DEHP on male reproduction.
Additional keywords: capacitation, human reproduction, phthalate ester, progesterone.
References
Alasmari, W., Costello, S., Correia, J., Oxenham, S. K., Morris, J., Fernandes, L., Ramalho-Santos, J., Kirkman-Brown, J., Michelangeli, F., Publicover, S., and Barratt, C. L. (2013). Ca2+ signals generated by CatSper and Ca2+ stores regulate different behaviors in human sperm. J. Biol. Chem. 288, 6248–6258.| Ca2+ signals generated by CatSper and Ca2+ stores regulate different behaviors in human sperm.Crossref | GoogleScholarGoogle Scholar | 23344959PubMed |
Caldwell, J. C. (2012). DEHP: genotoxicity and potential carcinogenic mechanisms – a review. Mutat. Res. 751, 82–157.
| DEHP: genotoxicity and potential carcinogenic mechanisms – a review.Crossref | GoogleScholarGoogle Scholar | 22484601PubMed |
Chang, W. H., Wu, M. H., Pan, H. A., Guo, P. L., and Lee, C. C. (2017). Semen quality and insulin-like factor 3: associations with urinary and seminal levels of phthalate metabolites in adult males. Chemosphere 173, 594–602.
| Semen quality and insulin-like factor 3: associations with urinary and seminal levels of phthalate metabolites in adult males.Crossref | GoogleScholarGoogle Scholar | 28152410PubMed |
Chen, J., Wu, S., Wen, S., Shen, L., Peng, J., Yan, C., Cao, X., Zhou, Y., Long, C., Lin, T., He, D., Hua, Y., and Wei, G. (2015). The mechanism of environmental endocrine disruptors (DEHP) induces epigenetic transgenerational inheritance of cryptorchidism. PLoS One 10, e0126403.
| The mechanism of environmental endocrine disruptors (DEHP) induces epigenetic transgenerational inheritance of cryptorchidism.Crossref | GoogleScholarGoogle Scholar | 26720722PubMed |
Frederiksen, H., Jørgensen, N., and Andersson, A. M. (2010). Correlations between phthalate metabolites in urine, serum, and seminal plasma from young Danish men determined by isotope dilution liquid chromatography tandem mass spectrometry. J. Anal. Toxicol. 34, 400–410.
| Correlations between phthalate metabolites in urine, serum, and seminal plasma from young Danish men determined by isotope dilution liquid chromatography tandem mass spectrometry.Crossref | GoogleScholarGoogle Scholar | 20822678PubMed |
Fredricsson, B., Möller, L., Pousette, Å., and Westerholm, R. (1993). Human sperm motility is affected by plasticizers and diesel particle extracts. Pharmacol. Toxicol. 72, 128–133.
| Human sperm motility is affected by plasticizers and diesel particle extracts.Crossref | GoogleScholarGoogle Scholar | 7682691PubMed |
Gao, D. W., and Wen, Z. D. (2016). Phthalate esters in the environment: a critical review of their occurrence, biodegradation, and removal during wastewater treatment processes. Sci. Total Environ. 541, 986–1001.
| Phthalate esters in the environment: a critical review of their occurrence, biodegradation, and removal during wastewater treatment processes.Crossref | GoogleScholarGoogle Scholar | 26473701PubMed |
Kasahara, E., Sato, E. F., Miyoshi, M., Konaka, R., Hiramoto, K., Sasaki, J., Tokuda, M., Nakano, Y., and Inoue, M. (2002). Role of oxidative stress in germ cell apoptosis induced by di(2-ethylhexyl)phthalate. Biochem. J. 365, 849–856.
| Role of oxidative stress in germ cell apoptosis induced by di(2-ethylhexyl)phthalate.Crossref | GoogleScholarGoogle Scholar | 11982482PubMed |
La Rocca, C., Tait, S., Guerranti, C., Busani, L., Ciardo, F., Bergamasco, B., Perra, G., Mancini, F. R., Marci, R., Bordi, G., Caserta, D., Focardi, S., Moscarini, M., and Mantovani, A. (2015). Exposure to endocrine disruptors and nuclear receptors gene expression in infertile and fertile men from Italian areas with different environmental features. Int. J. Environ. Res. Public Health 12, 12426–12445.
| Exposure to endocrine disruptors and nuclear receptors gene expression in infertile and fertile men from Italian areas with different environmental features.Crossref | GoogleScholarGoogle Scholar | 26445054PubMed |
Li, S., Dai, J., Zhang, L., Zhang, J., Zhang, Z., and Chen, B. (2011). An association of elevated serum prolactin with phthalate exposure in adult men. Biomed. Environ. Sci. 24, 31–39.
| An association of elevated serum prolactin with phthalate exposure in adult men.Crossref | GoogleScholarGoogle Scholar | 21440837PubMed |
Li, X., Fang, E. F., Scheibye-Knudsen, M., Cui, H., Qiu, L., Li, J., He, Y., Huang, J., Bohr, V. A., Ng, T. B., and Guo, H. (2014). Di-(2-ethylhexyl) phthalate inhibits DNA replication leading to hyperPARylation, SIRT1 attenuation, and mitochondrial dysfunction in the testis. Sci. Rep. 4, 6434.
| Di-(2-ethylhexyl) phthalate inhibits DNA replication leading to hyperPARylation, SIRT1 attenuation, and mitochondrial dysfunction in the testis.Crossref | GoogleScholarGoogle Scholar | 25242624PubMed |
Naz, R. K., and Rajesh, P. B. (2004). Role of tyrosine phosphorylation in sperm capacitation/acrosome reaction. Reprod. Biol. Endocrinol. 2, 75.
| Role of tyrosine phosphorylation in sperm capacitation/acrosome reaction.Crossref | GoogleScholarGoogle Scholar | 15535886PubMed |
Pant, N., Shukla, M., Kumar Patel, D., Shukla, Y., Mathur, N., Kumar Gupta, Y., and Saxena, D. K. (2008). Correlation of phthalate exposures with semen quality. Toxicol. Appl. Pharmacol. 231, 112–116.
| Correlation of phthalate exposures with semen quality.Crossref | GoogleScholarGoogle Scholar | 18513777PubMed |
Pant, N., Pant, A., Shukla, M., Mathur, N., Gupta, Y., and Saxena, D. (2011). Environmental and experimental exposure of phthalate esters: the toxicological consequence on human sperm. Hum. Exp. Toxicol. 30, 507–514.
| Environmental and experimental exposure of phthalate esters: the toxicological consequence on human sperm.Crossref | GoogleScholarGoogle Scholar | 20551087PubMed |
Richburg, J. H., and Boekelheide, K. (1996). Mono-(2-ethylhexyl) phthalate rapidly alters both Sertoli cell vimentin filaments and germ cell apoptosis in young rat testes. Toxicol. Appl. Pharmacol. 137, 42–50.
| Mono-(2-ethylhexyl) phthalate rapidly alters both Sertoli cell vimentin filaments and germ cell apoptosis in young rat testes.Crossref | GoogleScholarGoogle Scholar | 8607140PubMed |
Schiffer, C., Müller, A., Egeberg, D. L., Alvarez, L., Brenker, C., Rehfeld, A., Frederiksen, H., Wäschle, B., Kaupp, U. B., Balbach, M., Wachten, D., Skakkebaek, N. E., Almstrup, K., and Strünker, T. (2014). Direct action of endocrine disrupting chemicals on human sperm. EMBO Rep. 15, 758–765.
| Direct action of endocrine disrupting chemicals on human sperm.Crossref | GoogleScholarGoogle Scholar | 24820036PubMed |
Silva, M. J., Samandar, E., Preau, J. L., Needham, L. L., and Calafat, A. M. (2006). Urinary oxidative metabolites of di(2-ethylhexyl) phthalate in humans. Toxicology 219, 22–32.
| Urinary oxidative metabolites of di(2-ethylhexyl) phthalate in humans.Crossref | GoogleScholarGoogle Scholar | 16332407PubMed |
Stenz, L., Escoffier, J., Rahban, R., Nef, S., and Paoloni-Giacobino, A. (2017). Testicular dysgenesis syndrome and long-lasting epigenetic silencing of mouse sperm genes involved in the reproductive system after prenatal exposure to DEHP. PLoS One 12, e0170441.
| Testicular dysgenesis syndrome and long-lasting epigenetic silencing of mouse sperm genes involved in the reproductive system after prenatal exposure to DEHP.Crossref | GoogleScholarGoogle Scholar | 28085963PubMed |
Teil, M. J., Blanchard, M., and Chevreuil, M. (2006). Atmospheric fate of phthalate esters in an urban area (Paris–France). Sci. Total Environ. 354, 212–223.
| Atmospheric fate of phthalate esters in an urban area (Paris–France).Crossref | GoogleScholarGoogle Scholar | 16398997PubMed |
Tian, M., Liu, L., Zhang, J., Huang, Q., and Shen, H. (2019). Positive association of low-level environmental phthalate exposure with sperm motility was mediated by DNA methylation: a pilot study. Chemosphere 220, 459–467.
| Positive association of low-level environmental phthalate exposure with sperm motility was mediated by DNA methylation: a pilot study.Crossref | GoogleScholarGoogle Scholar | 30594797PubMed |
Wang, Y., Liu, W., Yang, Q., Yu, M., and Zhang, Z. (2015). Di(2-ethylhexyl) phthalate exposure during pregnancy disturbs temporal sex determination regulation in mice offspring. Toxicology 336, 10–16.
| Di(2-ethylhexyl) phthalate exposure during pregnancy disturbs temporal sex determination regulation in mice offspring.Crossref | GoogleScholarGoogle Scholar | 26219507PubMed |
Wang, Y. X., Zeng, Q., Sun, Y., Yang, P., Wang, P., Li, J., Huang, Z., You, L., Huang, Y. H., Wang, C., Li, Y. F., and Lu, W. Q. (2016). Semen phthalate metabolites, semen quality parameters and serum reproductive hormones: a cross-sectional study in China. Environ. Pollut. 211, 173–182.
| Semen phthalate metabolites, semen quality parameters and serum reproductive hormones: a cross-sectional study in China.Crossref | GoogleScholarGoogle Scholar | 26766535PubMed |
World Health Oraganization (WHO) (2010). WHO laboratory manual for the examination and processing of human semen. Fifth edition. Available at http://www.who.int/reproductivehealth/publications/infertility/9789241547789/en/ [verified 11 December 2019].
Xie, F., Chen, X., Weng, S., Xia, T., Sun, X., Luo, T., and Li, P. (2019). Effects of two environmental endocrine disruptors di-n-butyl phthalate (DBP) and mono-n-butyl phthalate (MBP) on human sperm functions in vitro. Reprod. Toxicol. 83, 1–7.
| Effects of two environmental endocrine disruptors di-n-butyl phthalate (DBP) and mono-n-butyl phthalate (MBP) on human sperm functions in vitro.Crossref | GoogleScholarGoogle Scholar | 30391722PubMed |
You, L., Wang, Y.-X., Zeng, Q., Li, M., Huang, Y.-H., Hu, Y., Cao, W.-C., Liu, A.-L., and Lu, W.-Q. (2015). Semen phthalate metabolites, spermatozoa apoptosis, and DNA damage: a cross-sectional study in China. Environ. Sci. Technol. 49, 3805–3812.
| Semen phthalate metabolites, spermatozoa apoptosis, and DNA damage: a cross-sectional study in China.Crossref | GoogleScholarGoogle Scholar | 25671248PubMed |
Yuh, I. S., Cheong, H. T., Kim, J. T., Park, I. C., Park, C. K., and Yang, B. K. (2013). Effects of endocrine disruptors (NP, DBP and BPA) on sperm characteristics and development of IVF embryos in pig. J. Anim. Sci. Technol. 55, 237–247.
| Effects of endocrine disruptors (NP, DBP and BPA) on sperm characteristics and development of IVF embryos in pig.Crossref | GoogleScholarGoogle Scholar |
Yuwatini, E., Hata, N., and Taguchi, S. (2006). Behavior of di(2-ethylhexyl) phthalate discharged from domestic waste water into aquatic environment. J. Environ. Monit. 8, 191–196.
| Behavior of di(2-ethylhexyl) phthalate discharged from domestic waste water into aquatic environment.Crossref | GoogleScholarGoogle Scholar | 16395478PubMed |
Zou, Q. X., Peng, Z., Zhao, Q., Chen, H. Y., Cheng, Y. M., Liu, Q., He, Y. Q., Weng, S. Q., Wang, H. F., Wang, T., Zheng, L. P., and Luo, T. (2017). Diethylstilbestrol activates CatSper and disturbs progesterone actions in human spermatozoa. Hum. Reprod. 32, 290–298.
| Diethylstilbestrol activates CatSper and disturbs progesterone actions in human spermatozoa.Crossref | GoogleScholarGoogle Scholar | 28031325PubMed |
