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Embryonic exposures to mono-2-ethylhexyl phthalate induce larval steatosis in zebrafish independent of Nrf2a signaling

Published online by Cambridge University Press:  17 February 2020

Karilyn E. Sant Open the ORCID record for Karilyn E. Sant [Opens in a new window] ,
Hadley M. Moreau ,
Larissa M. Williams ,
Haydee M. Jacobs ,
Anna M. Bowsher ,
Jason D. Boisvert ,
Roxanna M. Smolowitz ,
Jacob Pantazis ,
Kate Annunziato  and
Malina Nguyen
...Show all authors
Karilyn E. Sant*
Affiliation:
Division of Environmental Health, School of Public Health, San Diego State University, San Diego92182, CA, USA Department of Environmental Health Sciences, School of Public Health and Health Sciences, University of Massachusetts, Amherst01003, MA, USA
Hadley M. Moreau
Affiliation:
Department of Environmental Health Sciences, School of Public Health and Health Sciences, University of Massachusetts, Amherst01003, MA, USA Department of Biology, Bates College, Lewiston04240, ME, USA
Larissa M. Williams
Affiliation:
Department of Biology, Bates College, Lewiston04240, ME, USA
Haydee M. Jacobs
Affiliation:
Department of Environmental Health Sciences, School of Public Health and Health Sciences, University of Massachusetts, Amherst01003, MA, USA
Anna M. Bowsher
Affiliation:
Department of Biology, Bates College, Lewiston04240, ME, USA
Jason D. Boisvert
Affiliation:
Department of Biology, Bates College, Lewiston04240, ME, USA
Roxanna M. Smolowitz
Affiliation:
Department of Biology, Roger Williams University, Bristol02809, RI, USA
Jacob Pantazis
Affiliation:
Department of Biology, Bates College, Lewiston04240, ME, USA
Kate Annunziato
Affiliation:
Department of Environmental Health Sciences, School of Public Health and Health Sciences, University of Massachusetts, Amherst01003, MA, USA
Malina Nguyen
Affiliation:
Department of Environmental Health Sciences, School of Public Health and Health Sciences, University of Massachusetts, Amherst01003, MA, USA
Alicia Timme-Laragy
Affiliation:
Department of Environmental Health Sciences, School of Public Health and Health Sciences, University of Massachusetts, Amherst01003, MA, USA
*
Address for correspondence: Karilyn Sant, School of Public Health, San Diego State University, San Diego, CA92128, USA. Email: [email protected]
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Abstract

Mono-2-ethylhexyl phthalate (MEHP) is the primary metabolite of the ubiquitous plasticizer and toxicant, di-2-ethylhexyl phthalate. MEHP exposure has been linked to abnormal development, increased oxidative stress, and metabolic syndrome in vertebrates. Nuclear factor, Erythroid 2 Like 2 (Nrf2), is a transcription factor that regulates gene expression in response to oxidative stress. We investigated the role of Nrf2a in larval steatosis following embryonic exposure to MEHP. Wild-type and nrf2a mutant (m) zebrafish embryos were exposed to 0 or 200 μg/l MEHP from 6 to either 96 (histology) or 120 hours post fertilization (hpf). At 120 hpf, exposures were ceased and fish were maintained in clean conditions until 15 days post fertilization (dpf). At 15 dpf, fish lengths and lipid content were examined, and the expression of genes involved in the antioxidant response and lipid processing was quantified. At 96 hpf, a subset of animals treated with MEHP had vacuolization in the liver. At 15 dpf, deficient Nrf2a signaling attenuated fish length by 7.7%. MEHP exposure increased hepatic steatosis and increased expression of peroxisome proliferator-activated receptor alpha target fabp1a1. Cumulatively, these data indicate that developmental exposure alone to MEHP may increase risk for hepatic steatosis and that Nrf2a does not play a major role in this phenotype.

Keywords

PhthalateMEHPNrf2embryozebrafishlipid accumulationsteatosisDOHaD
Type
Original Article
Information
Journal of Developmental Origins of Health and Disease , Volume 12 , Issue 1 , February 2021 , pp. 132 - 140
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Copyright
© The Author(s) 2020. Published by Cambridge University Press and the International Society for Developmental Origins of Health and Disease

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References

Phthalates Action Plan, U.S.E.P. Agency, Editor. 2012.Google Scholar
Benjamin, S, Masai, E, Kamimura, N, Takahashi, K, Anderson, RC, Faisal, PA. Phthalates impact human health: epidemiological evidences and plausible mechanism of action. J Hazard Mater. 2017; 340, 360–383.CrossRefGoogle ScholarPubMed
Wittassek, M, Angerer, J. Phthalates: metabolism and exposure. Int J Androl. 2008; 31, 131–138.Google ScholarPubMed
Latini, G, De Felice, C, Verrotti, A. Plasticizers, infant nutrition and reproductive health. Reprod Toxicol. 2004; 19, 27–33.CrossRefGoogle ScholarPubMed
Frederiksen, H, Skakkebaek, NE, Andersson, AM. Metabolism of phthalates in humans. Mol Nutr Food Res. 2007; 51, 899–911.CrossRefGoogle ScholarPubMed
Inada, H, Chihara, K, Yamashita, A, et al. Evaluation of ovarian toxicity of mono-(2-ethylhexyl) phthalate (MEHP) using cultured rat ovarian follicles. J Toxicol Sci. 2012; 37, 483–490.CrossRefGoogle ScholarPubMed
Tomita, I, Nakamura, Y, Yagi, Y, Tutikawa, K. Fetotoxic effects of mono-2-ethylhexyl phthalate (MEHP) in mice. Environ Health Perspect. 1986; 65, 249–254.Google ScholarPubMed
Guibert, E, Prieur, B, Cariou, R, et al. Effects of mono-(2-ethylhexyl) phthalate (MEHP) on chicken germ cells cultured in vitro. Environ Sci Pollut Res. 2013; 20, 2771–2783.Google ScholarPubMed
Jacobs, HM, Sant, KE, Basnet, A, Williams, LM, Moss, JB, Timme-Laragy, AR. Embryonic exposure to Mono(2-ethylhexyl) phthalate (MEHP) disrupts pancreatic organogenesis in zebrafish (Danio rerio). Chemosphere. 2018; 195, 498–507.CrossRefGoogle ScholarPubMed
Sant, KE, Dolinoy, DC, Jilek, JL, Sartor, MA, Harris, C. Mono-2-ethylhexyl phthalate disrupts neurulation and modifies the embryonic redox environment and gene expression. Reprod Toxicol. 2016; 63, 32–48.CrossRefGoogle ScholarPubMed
Swan, SH, Sathyanarayana, S, Barrett, ES, et al. First trimester phthalate exposure and anogenital distance in newborns. Human Reproduction (Oxford, England) 2015; 30, 963–972.CrossRefGoogle ScholarPubMed
Sathyanarayana, S, Barrett, E, Nguyen, R, Redmon, B, Haaland, W, Swan, SH. First trimester phthalate exposure and infant birth weight in the infant development and environment study. Int J Environ Res Public Health 2016; 13, 945.CrossRefGoogle ScholarPubMed
Harley, KG Berger, K, Rauch, S, et al. Association of prenatal urinary phthalate metabolite concentrations and childhood BMI and obesity. Pediatr Res. 2017; 82, 405–415.CrossRefGoogle ScholarPubMed
Heindel, JJ. History of the obesogen field: looking back to look forward. Front Endocrinol. 2019; 10, 14–14.CrossRefGoogle ScholarPubMed
Heindel, JJ, Blumberg, B. Environmental obesogens: mechanisms and controversies. Annu Rev Pharmacol Toxicol. 2019; 59, 89–106.CrossRefGoogle ScholarPubMed
Grün, F, Blumberg, B. Endocrine disrupters as obesogens. Mol Cell Endocrinol. 2009; 304, 19–29.CrossRefGoogle ScholarPubMed
Grün, F, Blumberg, B. Environmental obesogens: organotins and endocrine disruption via nuclear receptor signaling. Endocrinology. 2006; 147, s50–s55.CrossRefGoogle ScholarPubMed
Wang, W, Craig, ZR, Basavarajappa, MS, Hafner, KS, Flaws, JA. Mono-(2-ethylhexyl) phthalate induces oxidative stress and inhibits growth of mouse ovarian antral follicles. Biol Reprod. 2012; 87, 152–152.Google ScholarPubMed
Pi, J, Leung, L, Xue, P, et al. Deficiency in the nuclear factor E2-related Factor-2 transcription factor results in impaired adipogenesis and protects against diet-induced obesity. J Biol Chem. 2010; 285, 9292–9300.CrossRefGoogle ScholarPubMed
Sheikh, IA, Abu-Elmagd, M, Turki, RF, Damanhouri, GA, Beg, MA, Al-Qahtani, M. Endocrine disruption: in silico perspectives of interactions of di-(2-ethylhexyl)phthalate and its five major metabolites with progesterone receptor. BMC Struct Biol. 2016; 16(Suppl. 1), 16–16.CrossRefGoogle ScholarPubMed
Zhai, W, Huang, Z, Chen, L, Feng, C, Li, B, Li, T. Thyroid endocrine disruption in zebrafish larvae after exposure to mono-(2-Ethylhexyl) phthalate (MEHP). PLoS One. 2014; 9, e92465.CrossRefGoogle ScholarPubMed
Shelton, P, Jaiswal, AK. The transcription factor NF-E2-related factor 2 (Nrf2): a protooncogene? FASEB J. 2013; 27, 414–423.Google ScholarPubMed
Kwong, M, Kan, YW, Chan, JY. The CNC basic leucine zipper factor, Nrf1, is essential for cell survival in response to oxidative stress-inducing agents. Role for Nrf1 in gamma-gcs(l) and gss expression in mouse fibroblasts. J Biol Chem. 2000; 274; 37491–37498.CrossRefGoogle Scholar
Sykiotis, GP, Bohmann, D. Stress-activated cap’n’collar transcription factors in aging and human disease. Sci Signal. 2010; 3, re3–re3.CrossRefGoogle ScholarPubMed
Ma, Q. Role of nrf2 in oxidative stress and toxicity. Annu Rev Pharmacol Toxicol. 2013; 53, 401–426.CrossRefGoogle ScholarPubMed
Espinosa-Diez, C, Miguel, V, Mennerich, D, et al. Antioxidant responses and cellular adjustments to oxidative stress. Redox Biol. 2015; 6, 183–197.CrossRefGoogle ScholarPubMed
Timme-Laragy, AR, Karchner, SI, Franks, DG, et al. Nrf2b, novel zebrafish paralog of oxidant-responsive transcription factor NF-E2-related factor 2 (NRF2). J Biol Chem. 2012; 287, 4609–4627.Google ScholarPubMed
Kobayashi, M, Itoh, K, Suzuki, T, et al. Identification of the interactive interface and phylogenic conservation of the Nrf2-Keap1 system. Genes Cells. 2002; 7, 807–820.CrossRefGoogle ScholarPubMed
Sant, KE, et al. Nrf2a modulates the embryonic antioxidant response to perfluorooctanesulfonic acid (PFOS) in the zebrafish, Danio rerio. Aquat Toxicol. 2018; 198, 92–102.Google ScholarPubMed
Sant, KE, et al. The role of Nrf1 and Nrf2 in the regulation of glutathione and redox dynamics in the developing zebrafish embryo. Redox Biol. 2017; 13 (Suppl. C), 207–218.CrossRefGoogle ScholarPubMed
Furukawa, S, Fujita, T, Shimabukuro, M, et al. Increased oxidative stress in obesity and its impact on metabolic syndrome. J Clin Invest. 2004; 114, 1752–1761.CrossRefGoogle ScholarPubMed
Marseglia, L, Manti, S, D’Angelo, G, et al. Oxidative stress in obesity: a critical component in human diseases. Int J Mol Sci. 2014; 16, 378–400.CrossRefGoogle ScholarPubMed
Takahashi, T, et al. Carnosic acid and carnosol inhibit adipocyte differentiation in mouse 3T3-L1 cells through induction of phase2 enzymes and activation of glutathione metabolism. Biochem Biophys Res Commun. 2009; 382, 549–554.Google ScholarPubMed
Shin, S, Wakabayashi, J, Yates, MS, et al. Role of Nrf2 in prevention of high-fat diet-induced obesity by synthetic triterpenoid CDDO-Imidazolide. Eur J Pharmacol. 2009; 620, 138–144.CrossRefGoogle ScholarPubMed
Seo, H-A, Lee, I-K. The role of Nrf2: adipocyte differentiation, obesity, and insulin resistance. Oxid Med Cell Longevity. 2013; 2013, 184598–184598.CrossRefGoogle Scholar
Physiology, Liver. 2018 18 December 2018 [cited 4 March 2019]; Available from: https://www.ncbi.nlm.nih.gov/books/NBK535438/.Google Scholar
Benedict, M, Zhang, X. Non-alcoholic fatty liver disease: an expanded review. World J Hepatol. 2017; 9, 715–732.CrossRefGoogle Scholar
Sayiner, M, et al., Epidemiology of nonalcoholic fatty liver disease and nonalcoholic steatohepatitis in the United States and the rest of the world. Clin Liver Dis. 2016; 20, 205–214.Google ScholarPubMed
Arciello, M, Gori, M, Maggio, R, et al. Environmental pollution: a tangible risk for NAFLD pathogenesis. Int J Mol Sci. 2013; 14.CrossRefGoogle ScholarPubMed
Mukaigasa, K, et al. Genetic evidence of an evolutionarily conserved role for Nrf2 in the protection against oxidative stress. Mol Cell Biol. 2012; 32, 4455–4461.CrossRefGoogle ScholarPubMed
Rousseau, ME, et al. Regulation of Ahr signaling by Nrf2 during development: effects of Nrf2a deficiency on PCB126 embryotoxicity in zebrafish (Danio rerio). Aquat Toxicol. (Amsterdam, Netherlands) 2015; 167, 157–171.Google ScholarPubMed
Schlegel, A, Stainier, DYR. Microsomal triglyceride transfer protein is required for yolk lipid utilization and absorption of dietary lipids in zebrafish larvae. Biochemistry. 2006; 45, 15179–15187.CrossRefGoogle ScholarPubMed
Fraher, D, et al. Lipid abundance in zebrafish embryos is regulated by complementary actions of the endocannabinoid system and retinoic acid pathway. Endocrinology. 2015; 156, 3596–3609.CrossRefGoogle ScholarPubMed
Livak, KJ, Schmittgen, TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods. 2001; 25, 402–408.CrossRefGoogle Scholar
McCurley, AT, Callard, GVJBMB. Characterization of housekeeping genes in zebrafish: male-female differences and effects of tissue type, developmental stage and chemical treatment. BMC Mol Biol. 2008; 9, 102.CrossRefGoogle ScholarPubMed
Cooper, CA, Handy, RD, Bury, NR. The effects of dietary iron concentration on gastrointestinal and branchial assimilation of both iron and cadmium in zebrafish (Danio rerio). Aquat Toxicol. 2006; 79, 167–175.Google Scholar
Zhao, X, et al. Klf6/copeb is required for hepatic outgrowth in zebrafish and for hepatocyte specification in mouse ES cells. Dev Biol. 2010; 344, 79–93.CrossRefGoogle ScholarPubMed
Laprairie, RB, Denovan-Wright, EM, Wright, JM. Subfunctionalization of peroxisome proliferator response elements accounts for retention of duplicated fabp1 genes in zebrafish. BMC Evol Biol. 2016; 16, 147.CrossRefGoogle ScholarPubMed
Schultz, LE, et al. Epigenetic regulators Rbbp4 and Hdac1 are overexpressed in a zebrafish model of RB1 embryonal brain tumor, and are required for neural progenitor survival and proliferation. Dis Model Mech. 2018; 11, dmm034124.Google Scholar
Williams, LM, et al. Developmental expression of the Nfe2-related factor (Nrf) transcription factor family in the Zebrafish, Danio rerio. PLoS One. 2013; 8, e79574.CrossRefGoogle ScholarPubMed
Williams, LM, et al. The transcription factor, Nuclear factor, erythroid 2 (Nfe2), is a regulator of the oxidative stress response during Danio rerio development. Aquat Toxicol. (Amsterdam, Netherlands) 2016; 180, 141–154.CrossRefGoogle Scholar
Cederbaum, AI, Nrf2 and antioxidant defense against CYP2E1 toxicity. In Cytochrome P450 2E1: Its Role in Disease and Drug Metabolism (ed. Dey, A), 2013; pp. 105–130. Springer Netherlands, Dordrecht.CrossRefGoogle Scholar
Thisse, B, et al. Expression of the zebrafish genome during embryogenesis (NIH R01 RR15402), in ZFIN Direct Data Submission (http://zfin.org). 2001.Google Scholar
Fagerberg, L, et al. Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics. Mol Cell Proteomics. 2014; 13, 397–406.CrossRefGoogle ScholarPubMed
Feige, JN, et al. The endocrine disruptor monoethyl-hexyl-phthalate is a selective peroxisome proliferator-activated receptor γ modulator that promotes adipogenesis. J Biol Chem. 2007; 282, 19152–19166.CrossRefGoogle ScholarPubMed
Hurst, CH, Waxman, DJ. Activation of PPARα and PPARγ by environmental phthalate monoesters. Toxicol Sci. 2003; 74, 297–308.CrossRefGoogle ScholarPubMed
Sant, KE, et al. Embryonic exposures to perfluorooctanesulfonic acid (PFOS) disrupt pancreatic organogenesis in the zebrafish, Danio rerio. Environ Pollut. 2017; 220, 807–817.Google ScholarPubMed
Fenton, TR, Kim, JH. A systematic review and meta-analysis to revise the Fenton growth chart for preterm infants. BMC Pediatr. 2013; 13, 59.CrossRefGoogle ScholarPubMed
Dulloo, AG, et al. The thrifty ‘catch-up fat’ phenotype: its impact on insulin sensitivity during growth trajectories to obesity and metabolic syndrome. Int J Obes. 2006; 30, S23–S35.CrossRefGoogle ScholarPubMed
Vaag, A. Low birth weight and early weight gain in the metabolic syndrome: consequences for infant nutrition. Int J Gynecol Obstet. 2009; 104 (Supplement), S32–S34.Google ScholarPubMed
Simmons, R. Developmental origins of adult metabolic disease: concepts and controversies. Trends Endocrinol Metabol. 2005; 16, 390–394.CrossRefGoogle ScholarPubMed
Pan, L, et al. Trends in severe obesity among children aged 2 to 4 years enrolled in special supplemental nutrition program for women, infants, and children from 2000 to 2014 trends in severe obesity among US children aged 2 to 4 years enrolled in WICTrends in severe obesity among US children aged 2 to 4 years enrolled in WIC. JAMA Pediatr. 2018; 172, 232–238.CrossRefGoogle Scholar
Fryar, CD, Carroll, MD, Ogden, CL. Prevalence of Overweight and Obesity among Children and Adolescents: United States, 1963–1965 Through 2011–2012, N.C.f.H. Statistics, Editor. 2014, Centers for Disease Control and Prevention, Division of Health and Nutrition Examination Surveys. Hyattsville, MD.Google Scholar
Bai, J, et al. Mono-2-ethylhexyl phthalate induces the expression of genes involved in fatty acid synthesis in HepG2 cells. Environ Toxicol Pharmacol. 2019; 69, 104–111.CrossRefGoogle ScholarPubMed
Zhang, Y, et al. Mono-2-ethylhexyl phthalate (MEHP) promoted lipid accumulation via JAK2/STAT5 and aggravated oxidative stress in BRL-3A cells. Ecotoxicol Environ Saf. 2019; 184, 109611.Google ScholarPubMed
Pereira, C, Mapuskar, K, Rao, CV. Chronic toxicity of diethyl phthalate in male Wistar rats – a dose–response study. Regul Toxicol Pharm. 2006; 45, 169–177.CrossRefGoogle ScholarPubMed
Ibabe, A, Bilbao, E, Cajaraville, MP. Expression of peroxisome proliferator-activated receptors in zebrafish (Danio rerio) depending on gender and developmental stage. Histochem Cell Biol. 2005; 123, 75–87.CrossRefGoogle ScholarPubMed
Liss, KHH, Finck, BN. PPARs and nonalcoholic fatty liver disease. Biochimie. 2017; 136, 65–74.CrossRefGoogle ScholarPubMed
Liew, WC, et al. Polygenic sex determination system in zebrafish. PLoS One. 2012; 7, e34397–e34397.CrossRefGoogle ScholarPubMed
Uchida, D, et al. Oocyte apoptosis during the transition from ovary-like tissue to testes during sex differentiation of juvenile zebrafish. J Exp Biol. 2002; 205, 711–718.Google ScholarPubMed
Phillips, ML. Phthalates and metabolism: exposure correlates with obesity and diabetes in men. Environ Health Perspect. 2007; 115, A312–A312.Google Scholar
Ballestri, S, et al. NAFLD as a sexual dimorphic disease: role of gender and reproductive status in the development and progression of nonalcoholic fatty liver disease and inherent cardiovascular risk. Adv Therapy. 2017; 34, 1291–1326.CrossRefGoogle ScholarPubMed
Bertolotti, M, et al. Nonalcoholic fatty liver disease and aging: epidemiology to management. World J Gastroenterol. 2014; 20, 14185–14204.CrossRefGoogle Scholar
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