Differential Effects of Bisphenol A and Di (2-Ethylhexyl) Phthalate Exposure on Crestin and the Expression of Some Genes Related to Apoptosis and Inflammation in Zebrafish Embryos

Year 2021, Volume: 80 Issue: 1, 42 - 47, 15.06.2021

Perihan Seda Ateş-kalkan Tuğçe Ayık Ünsal Veli Üstündağ Ebru Emekli Alturfan

Abstract

Objective: Endocrine disrupting chemicals (EDC) in plastics may disrupt proper endocrine system functioning. Zebrafish embryos are formed through external fertilization, and their rapid development, short life cycle, and transparency provide imaging advantages. In zebrafish embryos, neural crest development occurs similarly to other vertebrate embryos and crestin is found in the neural crest during embryogenesis. The aim of our study is to evaluate the effects of bisphenol A (BPA) and Di (2-ethylhexyl) phthalate (DEHP), which are the most widely used EDCs, on the expression of crestin, apoptosis, and inflammation-related parameters in zebrafish embryos.

Materials and Methods: The embryos were exposed to either DEHP or BPA in well plates for 72 h post fertilization (hpf ). Expressions of crestin were evaluated by in situ hybridization, while the expressions of bax, casp8, casp3a, ifng1, fas, and tp53 were evaluated by RT-PCR.

Results: Expressions of bax and casp8 increased and casp3a, ifng1, and fas decreased in BPA and DEHP groups. tp53 expression increased in the BPA group but decreased in the DEHP group compared with the control group. In the DEHP group, casp3a, ifng1, fas, bax, casp8, and tp53 expressions decreased compared with the BPA group. No significant change was observed in the crestin expressions in the groups. When compared with the control group, an inverse relation between ifng1 expression and apoptosis, as evidenced by increased bax and casp8 expressions, was observed in the BPA and DEHP groups.

Conclusion: Our study provided important data on the effects of EDCs on the relationship between inflammation and apoptosis.

Keywords

Bisphenol A,, Di (2-ethylhexyl) phthalate, Apoptosis, Inflammation, Crestin

References

• 1- Alabi OA, Ologbonjaye KI, Awosolu O, Alalade OE.Public and Environmental Health Effects of Plastic Wastes Disposal: A Review. J Toxicol Risk Assess 2019; 5:021.
• 2- Sax L. Polyethylene terephthalate may yield endocrine disruptors. Environ Health Persp 2010;118(4):445-8.
• 3- Durusoy R, Karababa AO. Plastik gıda ambalajları ve sağlık. TAF Prevent Med Bull 2011; 10(1): 87-96.
• 4- Strakovsky RS, Schantz SL. Impacts of bisphenol A (BPA) and phthalate exposures on epigenetic outcomes in the human placenta. Environ Epigenet 2018;4(3):dvy022.
• 5- Ustundag U , Unal İ , Ates P , Alturfan A , Yigitbasi T , Emekli-Alturfan E. Oxidant-Antioxidant Status and c-myc Expression in BPA- and DEHP-Exposed Zebrafish Embryos. Eur J Biol 2017; 76 (1): 26-30.
• 6- Mudbhary R, Sadler KC. Epigenetics, development, and cancer: zebrafish make their mark. Birth Defects Res C Embryo Today 2011; 93(2): 194-203.
• 7- Westerfield M. The Zebrafish Book, A Guide for the Laboratory Use of Zebrafish. University of Oregon Press, Oregon, USA (1995).
• 8- Lee YM, Rhee JS, Hwang DS, Kim IC, Raisuddin S, Lee JS. p53 gene expression is modulated by endocrine disrupting chemicals in the hermaphroditic fish, Kryptolebias marmoratus. Comp Biochem Physiol C Toxicol Pharmacol 2008;147(2):150-7.
• 9- Diamanti-Kandarakis E, Bourguignon JP, Giudice LC, et al. Endocrine-disrupting chemicals: an Endocrine Society scientific statement. Endocr Rev 2009;30(4):293-342.
• 10- Jocsak G, Ioja E, Kiss DS, Toth I, Barany Z, Bartha T, Frenyo LV, Zsarnovszky A. Endocrine Disruptors Induced Distinct Expression of Thyroid and Estrogen Receptors in Rat versus Mouse Primary Cerebellar Cell Cultures. Brain Sci 2019;5;9(12):359.
• 11- Anway MD, Skinner MK. Transgenerational effects of the endocrine disruptor vinclozolin on the prostate transcriptome and adult onset disease. Prostate 2008;68(5):517-529.
• 12- Üstündağ ÜV, Ünal İ, Ateş PS, Alturfan AA, Yiğitbaşı T, Emekli-Alturfan E. Bisphenol A and di(2-ethylhexyl) phthalate exert divergent effects on apoptosis and the Wnt/β-catenin pathway in zebrafish embryos: A possible mechanism of endocrine disrupting chemical action.Toxicol Ind Health 2017;33(12):901-910.
• 13- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25(4): 402-8.
• 14- Thisse C, Thisse B. High resolution in situ hybridization on whole-mount zebrafish embryo. Nat Protoc 2008; 3 : 59 – 69.
• 15- Lakind JS, Naiman DQ. Daily intake of bisphenol A and potential sources of exposure: 2005–2006 National Health and Nutrition Examination Survey. J Expo Sci Environ Epidemiol 2011; 21: 272–279.
• 16- Valentino R, D'Esposito V, Passaretti F et al. Bisphenol‐A impairs insulin action and up‐regulates inflammatory pathways in human subcutaneous adipocytes and 3T3‐L1 cells. PLoS ONE 2013;8: e82099.
• 17- Shamas-Din A, Kale J, Leber B, Andrews DW. Mechanisms of action of Bcl-2 family proteins. Cold Spring Harb Perspect Biol 2013;5(4):a008714.
• 18- Oka T, Adati N, Shinkai T, Sakuma K, Nishimura T, Kurose K. Bisphenol A induces apoptosis in central neural cells during early development of Xenopus laevis. Biochem Biophys Res Commun 2003; 312(4):877-82.
• 19- Lloyd V, Morse M, Purakal B, Parker J, Benard P, Crone M, Pfiffner S, Szmyd M, Dinda S. Hormone-like effects of bisphenol A on p53 and estrogen receptor alpha in breast cancer cells, BioResearch Open Access 2019; 8:1, 169–184.
• 20- Jiang W, Zhao H, Zhang L, Wu B, Zha Z. Maintenance of mitochondrial function by astaxanthin protects against bisphenol A-induced kidney toxicity in rats. Biomedicine&Pharmacotherapy 2020; 121:109629.
• 21- Xu J, Osuga Y, Yano T, Morita Y, Tang X, Fujiwara T, Takai Y, Matsumi H, Koga K, Taketani Y, Tsutsumi O. Bisphenol A induces apoptosis and G2-to-M arrest of ovarian granulosa cells. Biochem Biophys Res Commun. 2002; ;292(2):456-62.
• 22- Karpuzoglu-Sahin E, Zhi-Jun Y, Lengi A, Sriranganathan N, Ansar Ahmed S. Effects of long-term estrogen treat- ment on IFN-gamma, IL-2 and IL-4 gene expression and protein synthesis in spleen and thymus of normal C57BL/6 mice. Cytokine 2001; 14:208–217.
• 23- Sawai C, Anderson K, Walser-Kuntz D. Effect of Bisphenol A on Murine Immune Function: Modulation of Interferon-γ, IgG2a, and Disease Symptoms in NZB × NZW F1 Mice. Environmental Health Perspectives 2003; 111 (16).
• 24- Chen X, Xu S, Tan T, et al. Toxicity and estrogenic endocrine disrupting activity of phthalates and their mixtures. International Journal of Environmental Research and Public Health. 2014;11(3):3156–3168.
• 25- Li X, Fang E F, Scheibye-Knudsen M, et al. Di-(2-ethylhexyl) phthalate inhibits DNA replication leading to hyperPARylation, SIRT1 attenuation, and mitochondrial dysfunction in the testis. Scientific Reports 2014;4, 6434.
• 26- Wang W, Craig Z R, Basavarajappa MS, Gupta RK, Flaws JA. Di (2-ethylhexyl) phthalate inhibits growth of mouse ovarian antral follicles through an oxidative stress pathway. Toxicology and Applied Pharmacology. 2012;258(2):288–295.
• 27- Berke, N , Keklikoğlu, N. Nöral Krest Kökenli Hücrelerin Ağız ve Çene-Yüz Oluşumuna Katkıları ve Neden Oldukları Anomaliler. Journal of Istanbul University Faculty of Dentistry 2010; 44:39-43.
• 28- Mayor R, Theveneau E. The neural crest. Development 2013; 140: 2247-225.
• 29- Luo R, An M, Arduini BL, Henion PD. Specific pan-neural crest expression of zebrafish Crestin throughout embryonic development. Dev Dyn. 2001;220(2):169-74.