DOI QR코드

DOI QR Code

Effects of zearalenone on the localization and expression of the growth hormone receptor gene in the uteri of post-weaning piglets

  • Zhou, Min (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Yang, Li Jie (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Yang, Wei Ren (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Huang, Li Bo (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Zhou, Xue Mei (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Jiang, Shu Zhen (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University) ;
  • Yang, Zai Bin (Department of Animal Sciences and Technology and Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University)
  • Received : 2017.07.20
  • Accepted : 2017.09.15
  • Published : 2018.01.01

Abstract

Objective: In this study, we investigated the adverse effects of dietary zearalenone (ZEA) (0.5 to 1.5 mg/kg diet) on the localization and expression of the growth hormone receptor (GHR) in the uteri of post-weaning gilts and explored alternative mechanism of the reproductive toxicity of ZEA on piglets. Methods: A total of forty healthy piglets (Duroc${\times}$Landrace${\times}$Large White) aged 28 d were selected for study. Piglets were transferred to single cages after 10 days' adaptation on an obstetric table. The animals were allocated to one of four treatments: a normal basal diet supplemented with 0 (Control), 0.5 (ZEA0.5), 1.0 (ZEA1.0), or 1.5 (ZEA1.5) mg/kg purified ZEA, and fed for 35 d after the 10-d adaptation. Analyzed ZEA concentrations in the diets were 0, $0.52{\pm}0.07$, $1.04{\pm}0.03$, and $1.51{\pm}0.13mg/kg$, respectively. At the end of the feeding trial, piglets were euthanized after being fasted for 12 h. Two samples of uterine tissue from each pig were rapidly collected, one of which was stored at $-80^{\circ}C$ for analysis of the relative mRNA and protein expression of GHR, and the second was promptly fixed in Bouin's solution for immunohistochemical analysis. Results: The relative weight of the uteri and thickness of the myometrium and endometrium increased linearly (p<0.001) and quadratically (p<0.001) with an increasing level of ZEA. The results of immunohistochemical analysis indicated that GHR immunoreactive substance was mainly localizated in the cytoplasm of uterine smooth muscle, glandular epithelial, luminal epithelial, stromal, and vascular endothelial cells. In contrast, nuclear staining was rarely observed. The immunoreactive integrated optic density of GHR in the myometrium, luminal epithelium, glandular epithelium, and whole uteri of weaning gilts increased linearly (p<0.001) and quadratically (p<0.05) with an increasing level of ZEA. The mRNA and protein expression of GHR in the uteri of weaning gilts increased linearly (p<0.001) and quadratically (p<0.05) with an increasing level of ZEA. Conclusion: In conclusion, ZEA at a concentration of 0.5 mg/kg was sufficient to significantly thicken the myometrium and endometrium, and at a concentration of 1.0 mg/kg induced a high level of GHR expression to promote growth and development of the uteri. This revealed an alternative molecular mechanism whereby ZEA induces growth and development of the uteri and provides a theoretical basis for the revision of Chinese feed hygiene standards.

Keywords

References

  1. Huang LN, Peng SQ, Liu N. Research progress and development on reproductive toxicity of zearalenone. Chin J Contr Endem Dise 2014;29:181-4.
  2. Yazar S, Omurtag GZ. Fumonisins, trichothecenes and zearalenone in cereals. Int J Mol Sci 2008;9:2062-90. https://doi.org/10.3390/ijms9112062
  3. Denli M, Blandon JC, Salado S, Guynot ME, Perez JF. Effect of dietary zearalenone on the performance, reproduction tract and serum biochemistry in young rats. J Appl Anim Res 2017;45:619-22. https://doi.org/10.1080/09712119.2016.1251929
  4. Yang J, Zhang Y, Wang Y, Cui S. Toxic effects of zearalenone and alpha-zearalenol on the regulation of steroidogenesis and testosterone production in mouse Leydig cells. Toxicol In Vitro 2007;21:558-65.
  5. EFSA. Scientific opinion on the risks for public health related to the presence of zearalenone in food. EFSA J 2012;10:1-82.
  6. Bandera EV, Chandran U, Buckley B, et al. Urinary mycoestrogens, body size and breast development in New Jersey girls. Sci Total Environ 2011;409:5221-7. https://doi.org/10.1016/j.scitotenv.2011.09.029
  7. Li XY, Zhao LH, Fan Y, et al. Occurrence of mycotoxins in feed ingredients and complete feeds obtained from the Beijing region of China. J Anim Sci Biotechnol 2014;5:37. https://doi.org/10.1186/2049-1891-5-37
  8. Selvaraj JN, Wang Y, Zhou L, et al. Recent mycotoxin survey data and advanced mycotoxin detection techniques reported from China: a review. Food Addit Contam A 2015;32:440-52. https://doi.org/10.1080/19440049.2015.1010185
  9. EFSA. Opinion of the Scientific Panel on Contaminants in the Food Chain on a request from the Commission related to zearalenone as undesirable substance in animal feed. EFSA J 2004;89:1-35.
  10. EC. Commission recommendation of 17 August 2006: on the presence of deoxynivalenol, zearalenone, ochratoxin A, T-2 and HT-2 and fumonisin in products intended for animal feeding. Off J European Union 2006;229:7-9.
  11. Zou Y, Yang ZB, Yang WR, et al. Effect of purified zearalenone on nutrient digestibility in broilers fed 2 levels of fumonisin from naturally contaminated corn (Zea mays). J Appl Poult Res 2012;21:251-8. https://doi.org/10.3382/japr.2011-00359
  12. Heneweer M, Houtman R, Poortman J, et al. Estrogenic effects in the immature rat uteri after dietary exposure to ethinylestradiol and zearalenone using a systems biology approach. Toxicol Sci 2007; 99:303-14. https://doi.org/10.1093/toxsci/kfm151
  13. Cozzini P, Dellafiora L. In silico approach to evaluate molecular interaction between mycotoxins and the estrogen receptors ligand binding domain: a case study on zearalenone and its metabolites. Toxicol Lett 2012;214:81-5. https://doi.org/10.1016/j.toxlet.2012.07.023
  14. Jiang SZ, Yang ZB, Yang WR, et al. Effects of purified zearalenone on growth performance,organ size,serum metabolites, and oxidative stress in postweaning gilts. J Anim Sci 2011;89:3008-15. https://doi.org/10.2527/jas.2010-3658
  15. Etienne M, Jemmali M. Effects of zearalenone (F2) on estrous activity and reproduction in gilts. J Anim Sci 1982;55:1-10.
  16. Yakar S, Isaksson O. Regulation of skeletal growth and mineral acquisition by the GH/IGF-1 axis: Lessons from mouse models. Growth Horm IGF Res 2016;28:26-42. https://doi.org/10.1016/j.ghir.2015.09.004
  17. Sun F, Liu Y, Sun H, Tian B. Development and characterization of a novel GHR antibody antagonist, GF185. Int J Biol Macromol 2015; 79:864-70.
  18. Zeng C, Liu XL, Wang WM, et al. Characterization of GHRs, IGFs and MSTNs, and analysis of their expression relationships in blunt snout bream, Megalobrama amblycephala. Gene 2014;35:239-49.
  19. Gonzalez-Bulnes A, Lopez-Sebastian A, Garcia-Garcia RM, et al. Restoration of endocrine and ovarian function after stopping GnRH antagonist treatment in goats. Theriogenology 2005;63:83-91. https://doi.org/10.1016/j.theriogenology.2004.03.019
  20. Murphy LJ, Bell GI, Friesen HG. Tissue distribution of insulin-like growth factor-I and -II messenger ribonucleic acid in the adult rat. Eedocrinology 1987;120:1279-82. https://doi.org/10.1210/endo-120-4-1279
  21. Hafig A, Michel FJ, Simmen FA, Simmen RC. Constitutive expression of uterine receptors for insulin-like growth factor-I during the periimplantation period in the pig. Biol Reprod 1991;45:533-9.
  22. Taylor KM, Chen C, Gray AC, Bazer FW, Spencer TE. Expression of messenger ribonucleic acids for fibroblast growth factors 7 and 10 hepatocyte growth factor, and insulin-like growth factors and their receptors in the neonatal ovine uteri. Biol Reprod 2001;64:1236-46. https://doi.org/10.1095/biolreprod64.4.1236
  23. Chen XX, Yang WR, Zang CY, et al. Effects of fusarium toxins on growth performance, small intestinal disaccharidase activities and antioxidant capacity of weaning piglets. Chin J Anim Nutr 2015;27:1875-82.
  24. Dai ML, Jiang SZ, Yuan XJ, et al. Effects of zearalenone-diet on expression of Ghrelin and PCNA genes in ovaries of post-weaning piglets. Anim Reprod Sci 2016;168:126-37. https://doi.org/10.1016/j.anireprosci.2016.03.006
  25. NRC. Nutrient requirements of swine. Washington, DC, USA: National Academy; 2012.
  26. AOAC. Official Methods of Analysis of The AOAC. Rockville, MD, USA: AOAC Int.; 2012.
  27. Rivera A, Agnati LF, Horvath TL, et al. Uncoupling protein 2/3 immunoreactivity and theascending dopaminergic and noradrenergic neuronal systems:relevance for volume transmission. Neuroscience 2006;137:1447-61. https://doi.org/10.1016/j.neuroscience.2005.05.051
  28. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta C(T)) method. Methods 2001;25:402-8.
  29. Doll S, Gericke S, Danicke S, et al. The efficacy of a modified aluminosilicate as a detoxifying agent in Fusarium toxin contaminated maize containing diets for piglets. J Anim Physiol Anim Nutr 2005;89:342-58.
  30. Yang LJ, Niu QS, Zhang CY, et al. Changes in vulva, serum and oxidative stress index of fusarium toxin in weaning piglets. Swine Ind Sci 2016;33:85-8.
  31. Niu QS, Yang WR, Huang LB, et al. Effects of fusarium toxin on vulva, reproductive organ index, distribution and expression of estrogen receptors in uteri of weaning gilts. Chin J Anim Nutr 2016;28:1525-33.
  32. Jiang SZ, Yang ZB, Yang WR, et al. Effect of purified zearalenone with or without modified montmorillonite on nutrient availability, genital organs and serum hormones in post-weaning piglets. Livest Sci 2012;144:110-8. https://doi.org/10.1016/j.livsci.2011.11.004
  33. Powell JW, Raeford S, Lucier GW. Binding properties of zearalenone mycotoxins to hepatic estrogen receptors. Mol Pharmacol 1981;20:35-42.
  34. Alexopoulos C. Association of Fusarium mycotoxicosis with failure in applying an induction of parturition program with PGF2-alpha and oxytocin in sows. Theriogenology 2001;55:1745-57.
  35. Ojeda SR, Jameson HE. Developmental patterns of plasma and pituitary growth hormone (GH) in the female rat. Endocrinology 1977;100:881-9. https://doi.org/10.1210/endo-100-3-881
  36. Becci PJ, KA Voss, Hess FG, et al. Long-term carcinogenicity and toxicity study of zearalenone in the rat. J Appl Toxicol 1982;2:247-54. https://doi.org/10.1002/jat.2550020507
  37. Schoenmakers I, Kooistra HS, Okkens AC, et al. Growth hormone concentrations in mammary secretions and plasma of the periparturient bitch and in plasma of the neonate. J Reprod Fertil Supple 1997;51:363-7.
  38. Maranesi M, Parillo F, Leonardi L, Rebollar PG, et al. Expression of nerve growth factor and its receptors in the uterus of rabbits: functional involvement in prostaglandin synthesis. Domest Anim Endocrin 2016;56:20. https://doi.org/10.1016/j.domaniend.2016.02.001
  39. Hrabia A. Growth hormone production and role in the reproductive system of female chicken. Gen Comp Endocrinol 2015;220:112-8. https://doi.org/10.1016/j.ygcen.2014.12.022
  40. Sosa C, Carriquiry M, Chalar C, et al. Endometrial expression of leptin receptor and members of the growth hormone - insulin-like growth factor system throughout the estrous cycle in heifers. Anim Reprod Sci 2010;122:208-14. https://doi.org/10.1016/j.anireprosci.2010.08.011
  41. Zhu Qian. The expression of GH/IGF-I receptor and estrogen receptor in layers of uteri. Nanjing, China: Nanjing Agric Univ; 2003.
  42. Tong HN, Bai S, Wang SY, et al. Developmental changes in the distribution and expression of GHR and IGF-IR in the uteri of Jining Castle Peak sheep. Chin J Anim Vet Sci 2013;44:1392-9.
  43. Gajecka M. The effect of low-dose experimental zearalenone intoxication on the immunoexpression of estrogen receptors in the ovaries of pre-pubertal bitches. Pol J Vet Sci 2012;15:685-91.
  44. Wang DF, Zhou HL, Hou GY, Qi DS, Zhang NY. Soybean isoflavone reduces the residue of zearalenone in the muscle and liver of prepubertal gilts. Animal 2013;7:699-703. https://doi.org/10.1017/S1751731112002066
  45. Alm H, Greising T, Brussow KP, Torner H, Tieman U. The influence of the mycotoxins deoxynivalenol and zearalenol on in vitro maturation of pig oocytes and in vitro culture of pig zygotes. Toxi In Vitro 2002;16:643-8. https://doi.org/10.1016/S0887-2333(02)00059-0

Cited by

  1. Effects of Zearalenone Exposure on the TGF-β1/Smad3 Signaling Pathway and the Expression of Proliferation or Apoptosis Related Genes of Post-Weaning Gilts vol.10, pp.2, 2018, https://doi.org/10.3390/toxins10020049
  2. Current challenges in the diagnosis of zearalenone toxicosis as illustrated by a field case of hyperestrogenism in suckling piglets vol.4, pp.None, 2018, https://doi.org/10.1186/s40813-018-0095-4
  3. Comparative study of stress response, growth and development of uteri in post‐weaning gilts challenged with zearalenone and estradiol benzoate vol.103, pp.6, 2019, https://doi.org/10.1111/jpn.13195
  4. Illicium verum extracts and probiotics with added glucose oxidase promote antioxidant capacity through upregulating hepatic and jejunal Nrf2/Keap1 of weaned piglets vol.98, pp.3, 2018, https://doi.org/10.1093/jas/skaa077
  5. Effect of Illicium verum or Eucommia ulmoides leaf extracts on the anti‐stress ability, and mRNA and protein expression of Nrf2 and TNF‐α in Duroc × Landrace × Yorkshire vol.104, pp.4, 2020, https://doi.org/10.1111/jpn.13235
  6. Dose-Effect of Zearalenone on the Localization and Expression of Growth Hormone, Growth Hormone Receptor, and Heat Shock Protein 70 in the Ovaries of Post-weaning Gilts vol.8, pp.None, 2021, https://doi.org/10.3389/fvets.2021.629006
  7. Deoxynivalenol Induces Inflammation in the Small Intestine of Weaned Rabbits by Activating Mitogen-Activated Protein Kinase Signaling vol.8, pp.None, 2018, https://doi.org/10.3389/fvets.2021.632599
  8. Effect of Clostridium on proliferating cell nuclear antigen and ghrelin in the small intestine of fattening pigs fed with deoxynivalenol vol.14, pp.1, 2018, https://doi.org/10.3920/wmj2020.2569
  9. Impact of Fusarium-Derived Mycoestrogens on Female Reproduction: A Systematic Review vol.13, pp.6, 2018, https://doi.org/10.3390/toxins13060373
  10. Zearalenone exposure affects the Wnt/β-catenin signaling pathway and related genes of porcine endometrial epithelial cells in vitro vol.34, pp.6, 2018, https://doi.org/10.5713/ajas.20.0292