online ISSN 2415-3176
print ISSN 1609-6371
logoExperimental and Clinical Physiology and Biochemistry
  • 4 of 9
Up
ECPB 2020, 89(1): 29–35
https://doi.org/10.25040/ecpb2020.01.029
Research articles

High content of dietary fructose stimulates the formation of neutrophil extracellular traps in the biliary system

G.I. BILA1, O.I.VISHCHUR2, R.O. BILYY1
Abstract

Neutrophils safeguard our bloodstreams as well as the surface of ducts, mucosa linings and wounds, intending to protect us from pathogens. Bile ducts connect the bacteria-filled intestine with the liver, the metabolic factory of the body, full of sinusoidal capillaries. Thus, bile ducts become a hot place in the body, requiring constant protection from pathogens. Neutrophils can protect us in three different ways: they can phagocytize pathogens, produce chemicals that can mediate pathogens` death to form neutrophil extracellular traps (NETs). The latter are decondensed DNA fibers, decorated with toxic components of neutrophilic granules and usually associated with abundant ROS production. NETs are considered to be very effective in killing the bacteria, but, due to their nature, they are usually associated with inflammation. The latter can be self-limiting in physiological situations but upon the failure of the phagocytic system of our body or under the constant load of the trigger, it becomes chronic, causing deteriorating changes in the tissues. We demonstrated earlier that hydrophobic nanoparticles cause plasma membrane damage resulting in the NETs formation by neutrophils, it was also demonstrated that cholesterol nanocrystals can do the same. We have demonstrated recently that NETs are those factors initiating gall stone formation. Upon the diet containing high fructose content, there is an increased formation of monosodium urate microcrystals, they are well known for triggers of gout where NETs entrap MSU crystals. This data allowed us to assume the existence of the link between the high fructose content and the formation of NETs in bile ducts. To test our hypothesis we used the developed model of mouse NASH, using young growing mice C57BL6/N kept under normal ration, high fructose diet (HFD) and high lipid high cholesterol diet (HLHCD) as a recently found positive control for NETs formation. The fructose content was 10% in drinking water, roughly corresponding to the content of fructose in soft drinks consumed by humans. Before the beginning of the experiment and then every 14 days blood samples were collected, animals’ weight was monitored. After 6 weeks mice gall bladders were surgically removed and their contents were quantitatively transferred into glass slides and examined for the presence of NETs by visualizing DNA content with PI by means of the fluorescence microscopy. Under both HFD and HLHCD conditions we observed the formation of NETs in the gallbladder. Serum, obtained from blood samples, was analyzed for the activity of neutrophil elastase (an enzyme released from neutrophilic granules upon NETs formation) by using specific fluorogenic substrate. Neutrophil elastase activity was significantly increased after 6 weeks of feeding on the HFD (p=0.0219) and HLHCD (p=0.0148, ANOVA). Thus, we can conclude that high dietary fructose content stimulates the formation of neutrophil extracellular traps in bile ducts of experimental animals. These findings are extremely important in understanding the reasons for low-grade inflammation observed under non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

Recieved: 11.02.2020

Keywords: non-alcoholic steatohepatitis, neutrophil extracellular traps, NETs, gall bladder, inflammation

Full text: PDF (Ukr) 516K

References
  1. 1. Boeltz S, Amini P, Anders H-J, Andrade F, Bilyy R, Chatfield S, et al. To NET or not to NET:current opinions and state of the science regarding the formation of neutrophil extracellular traps. Cell Death Differ. 2019 [cited 2019 Jan 13];26(3):395-408. doi.org/10.1038/s41418-018-0261-x
  2. 2. Leppkes M, Schick M, Hohberger B, Mahajan A, Knopf J, Schett G, et al. Updates on NET formation in health and disease. Semin Arthritis Rheum. 2019;49(3):S43-8. doi.org/10.1016/j.semarthrit.2019.09.011
  3. 3. Leppkes M, Maueröder C, Hirth S, Nowecki S, Günther C, Billmeier U, et al. Externalized decondensed neutrophil chromatin occludes pancreatic ducts and drives pancreatitis. Nat Commun. 2016;7:10973. doi.org/10.1038/ncomms10973
  4. 4. Gunzer M. Escaping the traps of your own hunters. Science (80- ). 2017 [cited 2017 Dec 3];358(6367):1126-7. doi.org/10.1126/science.aar2428
  5. 5. Jiménez-Alcázar M, Rangaswamy C, Panda R, Bitterling J, Simsek YJ, Long AT, et al. Host DNases prevent vascular occlusion by neutrophil extracellular traps. Science. 2017 [cited 2017 Dec 3];358(6367):1202-6. doi.org/10.1126/science.aam8897
  6. 6. Muñoz LE, Bilyy R, Biermann MHC, Kienhöfer D, Maueröder C, Hahn J, et al. Nanoparticles size-dependently initiate self-limiting NETosis-driven inflammation. Proc Natl Acad Sci. 2016 [cited 2017 Nov 5];113(40):E5856-65. doi.org/10.1073/pnas.1602230113
  7. 7. Desai J, Foresto-Neto O, Honarpisheh M, Steiger S, Nakazawa D, Popper B, et al. Particles of different sizes and shapes induce neutrophil necroptosis followed by the release of neutrophil extracellular trap-like chromatin. Sci Rep. 2017;7(1):1-10& doi.org/10.1038/s41598-017-15106-0
  8. 8. Muñoz LE, Boeltz S, Bilyy R, Schauer C, Mahajan A, Widulin N, et al. Neutrophil Extracellular Traps Initiate Gallstone Formation. Immunity. 2019;51(3):443-450.e4. doi.org/10.1016/j.immuni.2019.07.002
  9. 9. Honda M, Kubes P. Neutrophils and neutrophil extracellular traps in the liver and gastrointestinal system. Nat Rev Gastroenterol Hepatol. 2018; doi.org/10.1038/nrgastro.2017.183
  10. 10. Hilscher MB, Shah VH. Neutrophil Extracellular Traps and Liver Disease. Semin Liver Dis. 2019 [cited 2019 Nov 19]; doi.org/10.1055/s-0039-3399562
  11. 11. Nat.Med.Editorial. Cutting out the liver fat. Nat Med. 2017;23(12):1385. doi.org/10.1038/nm.4459
  12. 12. Ronchi JA, Figueira TR, Ravagnani FG, Oliveira HCF, Vercesi AE, Castilho RF. A spontaneous mutation in the nicotinamide nucleotide transhydrogenase gene of C57BL/6J mice results in mitochondrial redox abnormalities. doi.org/10.1016/j.freeradbiomed.2013.05.049
  13. 13. Jensen T, Abdelmalek MF, Sullivan S, Nadeau KJ, Green M, Roncal C, et al. Fructose and sugar: A major mediator of non-alcoholic fatty liver disease. J Hepatol. 2018;68(5):1063-75. doi.org/10.1016/j.jhep.2018.01.019
  14. 14. Li M, Cai S-Y, Boyer JL. Mechanisms of bile acid mediated inflammation in the liver. Mol Aspects Med. 2017;56:45-53.doi.org/10.1016/j.mam.2017.06.001
  15. 15. Schauer C, Janko C, Munoz LE, Zhao Y, Kienhöfer D, Frey B, et al. Aggregated neutrophil extracellular traps limit inflammation by degrading cytokines and chemokines. Nat Med. 2014;20(5):511-7. doi.org/10.1038/nm.3547
  16. 16. Bila G, Peshkova S, Dumych T, Bilyy R. Natural cholesterol nanocrystals in gall material and their interaction with neutrophilic granulocytes. In: PhoBiA Annual Nanophotonics International Conference. Wroclaw; 2019. Р. 62.


Програмування - Roman.im