open access

Vol 56, No 2 (2018)
Original paper
Submitted: 2018-05-10
Accepted: 2018-06-02
Published online: 2018-06-08
Get Citation

Alterations in porcine intrahepatic sympathetic nerves after bisphenol A administration

Michael Thoene1, Janusz Godlewski2, Liliana Rytel3, Ewa Dzika1, Ewa Bejer-Olenska4, Joanna Wojtkiewicz45
DOI: 10.5603/FHC.a2018.0012
·
Pubmed: 29888781
·
Folia Histochem Cytobiol 2018;56(2):113-121.
Affiliations
  1. Department of Medical Biology, Faculty of Health Sciences, University of Warmia and Mazury in Olsztyn, Poland
  2. Department of Human Histology and Embryology, School of Medicine, Collegium Medicum, University of Warmia and Mazury in Olsztyn, Poland
  3. Department of Internal Medicine and Clinic, Faculty of Veterinary Medicine, University of Warmia and Mazury, Olsztyn, Poland
  4. Department of Pathophysiology, School of Medicine, Collegium Medicum, University of Warmia and Mazury, Olsztyn, Poland
  5. Laboratory of Regenerative Medicine. University of Warmia and Mazury in Olsztyn, Poland

open access

Vol 56, No 2 (2018)
ORIGINAL PAPERS
Submitted: 2018-05-10
Accepted: 2018-06-02
Published online: 2018-06-08

Abstract

 Introduction. Bisphenol A (BPA) is used in the chemical industry for manufacturing plastics which are used as food packaging. Data indicate that BPA is released from such products and is widely present in the environment and the human body. So far, the EFSA and the US FDA have determined “safe” BPA oral exposure levels, and a large amount of data indicates that BPA is harmful even at low-doses. Our previously performed analyses concerning BPA exposure demonstrated the impact of this substance on parasympathetic and peptidergic nerve fibers present within the liver. Therefore, this study concerns BPA exposure and sympathetic intrahepatic in­nervation in reference to several neuropeptides which modulate neuronal responses: cocaine and amphetamine regulated transcript (CART), galanin (GAL), calcitonin gene-regulated peptide (CGRP) and substance P (SP).

Materials and methods. Fifteen young swine at 8 weeks of age were used as experimental models of the juvenile human liver. The pigs were divided into 3 groups and received capsules orally with bisphenol at a dose of 0.05 mg/kg b.w./day; a dose of 0.5 mg/kg b.w./day and placebo capsules as a control. After 28 days of oral BPA intake, the animals were euthanized, perfused with 4% paraformaldehyde (PFA), and livers were collected and fixed in PFA. The cryostat sections were subjected to a routine double-labeling immunofluorescence technique. The primary antibodies were directed against dopamine beta hydroxylase (DbH), which is a marker for sympathetic nerves, and one of the investigated neuropeptides: CART, GAL, CGRP and SP, which co-localized the inves­tigated nerves. Immunoreactive nerves were counted in the liver and the percentage presence of each neuronal combination in particular samples of each experimental group were determined and analyzed statistically.

Results. The BPA oral intake at low and ten times higher dosage caused an increase of the number of sympa­thetic nerve fibers within the porcine liver by 48.6% and 63.7%, respectively. Moreover, BPA exposure caused an increased presence of sympathetic nerve fibers in these two experimental groups, which were co-localized with CART and GAL up to 65.9%/173.2% and 147.4%/126.3%, respectively. At the lower BPA doses of 50 μg/kg b.w./day, the percentages of SP+/DbH+ and CGRP+/DbH+ nerve fibers were similar to the control. However at a ten times higher dose, BPA caused an increased number of SP+/ DbH+ and CGRP+/ DbH+ nerve fibers in the liver, up to 46.4% and 73.5% respectively.

Conclusions. BPA caused an increase in the number of sympathetic nerve fibers as well as sympathetic nerve fibers which co-localized with neuropeptides in the porcine liver. The increase in CART and GAL were excep­tionally high even at low BPA doses. BPA food contamination may dysregulate liver sympathetic innervation, and thereby may change the oxygenated blood supply, alter metabolism and disrupt the activity of hepatic pa­renchymal cells.

Abstract

 Introduction. Bisphenol A (BPA) is used in the chemical industry for manufacturing plastics which are used as food packaging. Data indicate that BPA is released from such products and is widely present in the environment and the human body. So far, the EFSA and the US FDA have determined “safe” BPA oral exposure levels, and a large amount of data indicates that BPA is harmful even at low-doses. Our previously performed analyses concerning BPA exposure demonstrated the impact of this substance on parasympathetic and peptidergic nerve fibers present within the liver. Therefore, this study concerns BPA exposure and sympathetic intrahepatic in­nervation in reference to several neuropeptides which modulate neuronal responses: cocaine and amphetamine regulated transcript (CART), galanin (GAL), calcitonin gene-regulated peptide (CGRP) and substance P (SP).

Materials and methods. Fifteen young swine at 8 weeks of age were used as experimental models of the juvenile human liver. The pigs were divided into 3 groups and received capsules orally with bisphenol at a dose of 0.05 mg/kg b.w./day; a dose of 0.5 mg/kg b.w./day and placebo capsules as a control. After 28 days of oral BPA intake, the animals were euthanized, perfused with 4% paraformaldehyde (PFA), and livers were collected and fixed in PFA. The cryostat sections were subjected to a routine double-labeling immunofluorescence technique. The primary antibodies were directed against dopamine beta hydroxylase (DbH), which is a marker for sympathetic nerves, and one of the investigated neuropeptides: CART, GAL, CGRP and SP, which co-localized the inves­tigated nerves. Immunoreactive nerves were counted in the liver and the percentage presence of each neuronal combination in particular samples of each experimental group were determined and analyzed statistically.

Results. The BPA oral intake at low and ten times higher dosage caused an increase of the number of sympa­thetic nerve fibers within the porcine liver by 48.6% and 63.7%, respectively. Moreover, BPA exposure caused an increased presence of sympathetic nerve fibers in these two experimental groups, which were co-localized with CART and GAL up to 65.9%/173.2% and 147.4%/126.3%, respectively. At the lower BPA doses of 50 μg/kg b.w./day, the percentages of SP+/DbH+ and CGRP+/DbH+ nerve fibers were similar to the control. However at a ten times higher dose, BPA caused an increased number of SP+/ DbH+ and CGRP+/ DbH+ nerve fibers in the liver, up to 46.4% and 73.5% respectively.

Conclusions. BPA caused an increase in the number of sympathetic nerve fibers as well as sympathetic nerve fibers which co-localized with neuropeptides in the porcine liver. The increase in CART and GAL were excep­tionally high even at low BPA doses. BPA food contamination may dysregulate liver sympathetic innervation, and thereby may change the oxygenated blood supply, alter metabolism and disrupt the activity of hepatic pa­renchymal cells.

Get Citation

Keywords

BPA; intrahepatic innervation; sympathetic nervous system; DbH; CART; GAL; CGRP; SP

About this article
Title

Alterations in porcine intrahepatic sympathetic nerves after bisphenol A administration

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 56, No 2 (2018)

Article type

Original paper

Pages

113-121

Published online

2018-06-08

DOI

10.5603/FHC.a2018.0012

Pubmed

29888781

Bibliographic record

Folia Histochem Cytobiol 2018;56(2):113-121.

Keywords

BPA
intrahepatic innervation
sympathetic nervous system
DbH
CART
GAL
CGRP
SP

Authors

Michael Thoene
Janusz Godlewski
Liliana Rytel
Ewa Dzika
Ewa Bejer-Olenska
Joanna Wojtkiewicz

References (30)
  1. Tsai WT. Human health risk on environmental exposure to Bisphenol-A: a review. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev. 2006; 24(2): 225–255.
  2. Murata M, Kang JH. Bisphenol A (BPA) and cell signaling pathways. Biotechnol Adv. 2018; 36(1): 311–327.
  3. Vandenberg LN, Hauser R, Marcus M, et al. Human exposure to bisphenol A (BPA). Reprod Toxicol. 2007; 24(2): 139–177.
  4. Acconcia F, Pallottini V, Marino M. Molecular Mechanisms of Action of BPA. Dose Response. 2015; 13(4): 1559325815610582.
  5. European Food Safety Authority (EFSA) Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids. Scientific Opinion on the risks to public health related to the presence of bisphenol A (BPA) in foodstuffs: Executive summary. . ; 13(1): . https://. EFSA Journal 2015; 13(1): 3978 https://efsa onlinelibrary wiley com/doi/epdf/10 2903/j efsa 2015 3978 Accessed. ; 14: April.
  6. U.S. Food and Drug Administration. Bisphenol A (BPA). Final report for the review of literature and data on BPA 6/6/2014. https://www fda gov/Food/IngredientsPackagingLabeling/FoodAdditivesIngredients/ucm166145 htm Accessed. ; 14: April.
  7. Vandenberg LN, Colborn T, Hayes TB, et al. Hormones and endocrine-disrupting chemicals: low-dose effects and nonmonotonic dose responses. Endocr Rev. 2012; 33(3): 378–455.
  8. Welshons WV, Thayer KA, Judy BM, et al. Large effects from small exposures. I. Mechanisms for endocrine-disrupting chemicals with estrogenic activity. Environ Health Perspect. 2003; 111(8): 994–1006.
  9. Inadera H. Neurological Effects of Bisphenol A and its Analogues. Int J Med Sci. 2015; 12(12): 926–936.
  10. Kim K, Son TG, Kim SoJ, et al. Suppressive effects of bisphenol A on the proliferation of neural progenitor cells. J Toxicol Environ Health A. 2007; 70(15-16): 1288–1295.
  11. Szymanska K, Gonkowski S. Bisphenol A-Induced changes in the enteric nervous system of the porcine duodenum. Neurotoxicology. 2018; 66: 78–86.
  12. Szymanska K, Makowska K, Gonkowski S. The Influence of High and Low Doses of Bisphenol A (BPA) on the Enteric Nervous System of the Porcine Ileum. Int J Mol Sci. 2018; 19(3).
  13. Thoene M, Rytel L, Dzika E, et al. Bisphenol A Causes Liver Damage and Selectively Alters the Neurochemical Coding of Intrahepatic Parasympathetic Nerves in Juvenile Porcine Models under Physiological Conditions. Int J Mol Sci. 2017; 18(12).
  14. Thoene MA, Rytel L, Dzika E, et al. Immunohistochemical characteristics of porcine intrahepatic nerves under physiological conditions and after Bisphenol A administration. Folia Morphol (Warsz). 2018 [Epub ahead of print].
  15. Helke KL, Swindle MM. Animal models of toxicology testing: the role of pigs. Expert Opin Drug Metab Toxicol. 2013; 9(2): 127–139.
  16. Nykonenko A, Vávra P, Zonča P. Anatomic Peculiarities of Pig and Human Liver. Exp Clin Transplant. 2017; 15(1): 21–26.
  17. Wojtkiewicz J, Rytel L, Makowska K, et al. Co-localization of zinc transporter 3 (ZnT3) with sensory neuromediators and/or neuromodulators in the enteric nervous system of the porcine esophagus. Biometals. 2017; 30(3): 393–403.
  18. Lin YS, Nosaka S, Amakata Y, et al. Comparative study of the mammalian liver innervation: an immunohistochemical study of protein gene product 9.5, dopamine beta-hydroxylase and tyrosine hydroxylase. Comp Biochem Physiol A Physiol. 1995; 110(4): 289–298.
  19. Akiyoshi H, Gonda T, Terada T. A comparative histochemical and immunohistochemical study of aminergic, cholinergic and peptidergic innervation in rat, hamster, guinea pig, dog and human livers. Liver. 1998; 18(5): 352–359.
  20. Streba LA, Vere CC, Ionescu AG, et al. Role of intrahepatic innervation in regulating the activity of liver cells. World J Hepatol. 2014; 6(3): 137–143.
  21. Jensen KJ, Alpini G, Glaser S. Hepatic nervous system and neurobiology of the liver. Compr Physiol. 2013; 3(2): 655–665.
  22. Barja F, Mathison R. Sensory innervation of the rat portal vein and the hepatic artery. J Auton Nerv Syst. 1984; 10(2): 117–125.
  23. Berthoud HR. Anatomy and function of sensory hepatic nerves. Anat Rec A Discov Mol Cell Evol Biol. 2004; 280(1): 827–835.
  24. Schlereth T, Schukraft J, Krämer-Best HH, et al. Interaction of calcitonin gene related peptide (CGRP) and substance P (SP) in human skin. Neuropeptides. 2016; 59: 57–62.
  25. Kowalyk S, Veith R, Boyle M, et al. Liver releases galanin during sympathetic nerve stimulation. Am J Physiol. 1992; 262(5 Pt 1): E671–E678.
  26. Taborsky GJ, Dunning BE, Havel PJ, et al. The canine sympathetic neuropeptide galanin: a neurotransmitter in pancreas, a neuromodulator in liver. Horm Metab Res. 1999; 31(5): 351–354.
  27. Ekblad E. CART in the enteric nervous system. Peptides. 2006; 27(8): 2024–2030.
  28. Palus K, Rytel L. Co-localisation of cocaine- and amphetamine-regulated transcript peptide and vasoactive intestinal polypeptide in the myenteric plexus of the porcine transverse colon. Folia Morphologica. 2013; 72(4): 328–332.
  29. Wojtkiewicz J, Gonkowski S, Bladowski M, et al. Characterisation of cocaine- and amphetamine- regulated transcript-like immunoreactive (CART-LI) enteric neurons in the porcine small intestine. Acta Vet Hung. 2012; 60(3): 371–381.
  30. Wierup N, Gunnarsdóttir A, Ekblad E, et al. Characterisation of CART-containing neurons and cells in the porcine pancreas, gastro-intestinal tract, adrenal and thyroid glands. BMC Neurosci. 2007; 8: 51.

Regulations

Important: This website uses cookies. More >>

The cookies allow us to identify your computer and find out details about your last visit. They remembering whether you've visited the site before, so that you remain logged in - or to help us work out how many new website visitors we get each month. Most internet browsers accept cookies automatically, but you can change the settings of your browser to erase cookies or prevent automatic acceptance if you prefer.

By "Via Medica sp. z o.o." sp.k., ul. Świętokrzyska 73, 80–180 Gdańsk

tel.:+48 58 320 94 94, faks:+48 58 320 94 60, e-mail:  viamedica@viamedica.pl