Rats’ liver morphological features under conditions of different supply with vitamins and minerals
AbstractThe current practice of novel food safety assessment in the Russian Federation involves toxicological studies on the alimentary model of adaptation potential reduction of laboratory animals. Since vitamin and mineral deficiency can affect the size of structural elements of tissues, an objective estimation of the results obtained using this model is possible when determining the range of fluctuations of the studied morphometric parameters under conditions of different essential substances’ supply, as well as under conditions of simulated toxic effects on the background of the corresponding supply.
The purpose of the research was to investigate the morphological and morphometric features of the liver under the influence of reduced intake of vitamins and mineral elements in the combination with toxic effects of various nature, during growth and puberty of male Wistar rats.
Material and methods. The article analyzed data of 4 model experiments on 140 animals that received semi-synthetic casein diet with different supply of vitamins B1, B2, B3, B6 and mineral elements Fe3+ and Mg2+, as well as data of 2 experiments on 180 animals with simulated toxic load of cadmium (Cd2+) salts and carbon tetrachloride. The animals were ~95 days old at the time of sampling, the duration of the experiments was ~65 days. For the analysis we used data on rats’ body weight on the day of material sampling, absolute and relative liver weight, hepatocyte diameter, nucleus diameter and hepatocyte cytoplasm size in the central and peripheral zones of hepatic lobules. A total of 200 cells were analyzed in each group of animals. In accordance with the study design, all quantitative traits of the groups that received diets with an essential nutrient supply ranging from 75 to 2% were compared with the group that received a complete diet (100%).
Results. Morphometric examination of hepatocytes revealed a linear decrease in the size of cell structural elements in the series of reducing the content of essential micronutrients in the diet. Under the conditions of 2–4% vitamin and mineral supply, cell and nucleus diameters as well as cytoplasm size were by ~16.8, 12.6 and 21.1% (p<0.05) lower respectively than in rats with optimal supply of these substances; under the conditions of 9–19% supply were by ~9.2, 9.7 and 8.7% lower (p<0.05); higher levels of supply caused reduction of hepatocyte, nucleus and cytoplasm sizes in a range not exceeding 5% (p>0.05). When comparing the size of hepatocytes of rats subjected to toxic load with the hepatocytes of rats referred to the reference standard, an increase in the size of hepatocytes under the action of carbon tetrachloride by 17.4% (p<0.05) on average and under the action of cadmium salts by 4.6% (p<0.05) was noted.
Conclusion. Based on the analysis of liver morphological and morphometric studies’ data, there were established sizes of hepatocytes structural elements in the rats kept on diets with decreasing supply with B group vitamins, iron and magnesium salts; the linear decrease in the sizes of structural elements of hepatocytes in the series of reduction of B group vitamins, iron and magnesium intake was revealed. Toxic exposures to carbon tetrachloride and cadmium salts against the background of a 19–30–75% supply with essential substances led to an increase in the hepatocytes size, the correlation between the degree of toxicant exposure and the supply level is not significant.
Keywords:toxicological research; alimentary model of adaptation potential reducing; morphometry; hepatocytes’ structural elements; essential substances’ deficiency
Funding. The research was funded by the Russian Science Foundation (Project No. 20-16-00083-П).
Conflict of interest. The authors have no conflict of interest to declare.
Contribution. Сoncept and design of the study – Tyshko N.V.; collecting and statistical processing the material – Nikitin N.S.; text writing, editing, approval of the final version of the article, responsibility for the integrity of all parts of the article – all authors.
For citation: Nikitin N.S., Tyshko N.V. Rats’ liver morphological features under conditions of different supply with vitamins and minerals. Voprosy pitaniia [Problems of Nutrition]. 2023; 92 (5): 70–9. DOI: https://doi.org/10.33029/0042-8833-2023-92-5-70-79 (in Russian)
References
1. Tyshko N.V., Sadykova E.O., Timonin A.N., Shestakova S.I., Trebukh M.D., Pashorina V.A. Modification of vitamin-mineral diet composition as a model of adaptive potential reducing in laboratory animals. Voprosy pitaniia [Problems of Nutrition]. 2016; 85 (6): 64–71. DOI: https://doi.org/10.24411/0042-8833-2016-00079 (in Russian)
2. Tyshko N.V., Sadykova E.O., Timonin A.N., Shestakova S.I., Mustaphina O.K., Soto C.J. Research of the cadmium intoxication effect on the model of vitamin-mineral deficiency in rats. Voprosy pitaniia [Problems of Nutrition]. 2018; 87 (1): 63–71. DOI: https://doi.org/10.24411/0042-88332018-10007 (in Russian)
3. Nikitin N.S. Morphological changes in the liver of rats under conditions of vitamin-mineral deficiency. Voprosy pitaniia [Problems of Nutrition]. 2018; 87 (S5): 39–40. DOI: https://doi.org/10.24411/0042-8833-2018-10122 (in Russian)
4. Tsatsakis A., Goumenou M., Tyshko N.V., Shestakova S.I., Sadykova E.O., Zhminchenko V.M., et al. Detrimental effects of 6 months exposure to very low doses of a mixture of six pesticides associated with chronic vitamin deficiency on rats. Food Chem Toxicol. 2021; 152: 112188. DOI: https://doi.org/10.1016/j.fct.2021.112188
5. Tyshko N.V., Sadykova E.O., Shestakova S.I., Nikitin N.S., Trebukh M.D., et al. The use of the adaptation potential reduction model for reproductive toxicity research in vivo // J. Toxicol. 2020. Vol. 2020. Article ID 8834630. DOI: https://doi.org/10.1155/2020/8834630
6. Tyshko N.V., Nikitin N.S., Shestakova S.I., Sadykova E.O., Trebukh M.D., Guseva G.V. System biomarkers and liver morphology of rats during chronic low doses toxicants administration on the background of vitamin deficiency. Byulleten’ eksperimental’noi biologii i meditsiny [Bulletin of Experimental Biology and Medicine]. 2022; 174 (9): 356–60. DOI: https://doi.org/10.47056/0365-9615-2022-174-9-356-360 (in Russian)
7. Tyshko N.V., Shestakova S.I. Model of vitamin and mineral deficiency for toxicological research: apoptosis activity under conditions of CCL4 intoxication. Toxicol Rep. 2018; 6: 151–4. DOI: https://doi.org/10.1016/j.toxrep.2018.12.005
8. Nikitin N.S., Kuznetsov S.L., Tyshko N.V. Rats’ liver morphology under the conditions of CCL4 administration on the background of vitamin-mineral deficiency. Morfologiya [Morphology]. 2019; 155 (3): 42–7. (in Russian)
9. Korzhevsky D.E. Morphological diagnostics. Preparation of material for histological examination and electron microscopy. Saint Petersburg: SpetsLit, 2013: 128 p. ISBN 978-5-299-00569-1. (in Russian)
10. Bhadoria P., Nagar M., Bahrioke V., Bhadoria A. S. Effect of ethephon on the liver in albino rats: a histomorphometric study. Biomed J. 2015; 38 (5): 421–7. DOI: https://doi.org/10.4103/2319-4170.155589
11. Yanko R.V., Chaka E.G., Zinchenko A.S., Safonov S.L., Levashov M.I. Features of modeling fatty liver disease in rats of different ages based on a high-calorie diet. Ozhirenie i metabolizm [Obesity and Metabolism]. 2021; 18 (4): 387–97. DOI: https://doi.org/10.14341/omet12789 (in Russian)
12. Rosioru C., Talu S., Talu M., Giovanzana S., Craciun C. Morphometric assessments for the healthy rat hepatocytes. Ann RSCB. 2012; 17 (1): 74–9.
13. Prawitasari S., Saraswati T.R., Tana S. Liver histological structure of rats (Rattus norvegicus) in the lactation period after supplemented with organic quail eggs. J Phys Conf Ser. 2019; 1217: 012154. DOI: https://doi.org/10.1088/1742-6596/1217/1/012154
14. Yanko R.V. , Chaka E.G., Litovka I.G., Levashov M.I. Zhurnal Belorusskogo gosudarstvennovo universiteta. Biologiya [Journal of the Belarusian State University. Biology]. 2019; (3): 40 – 8. DOI: https://doi.org/10.33581/2521-1722-2019-3-40-48 (in Russian)
15. Yanko R., Chaka E., Levashov M. Morphofunctional changes in the rat’s liver of different ages after L-methionine administration. J Educ Health Sport. 2022; 12 (1): 125–37. DOI: https://doi.org/10.12775/JEHS.2022.12.01.010
16. Lebedeva E.I., Myadelets O.D., Prudnikov V.S. Morphofunctional characteristics of the white rats’ liver in norm. Uchenye zapiski UO VGAVM [Scientific Notes of the Educational Establishment «Vitebsk the Order of “the Badge of Honor”» State Academy of Veterinary Medicine]. 2015; 51 (1): 80–3. (in Russian)
17. El’chaninov A.V., Bol’shakova G.B. Size of hepatocytes and their nuclei in regenerating fetal rats’ liver. Rossiyskiy mediko-biologicheskiy vestnik imeni akademika I.P. Pavlova [Russian Medical and Biological Bulletin named after academician I.P. Pavlov]. 2011; 19 (2): 1–4. (in Russian)
18. Dzhadranov E.S., Ergazina M.Zh., Dzhangel’dina Z.N., Krasnoshtanov A.V., Krasnoshtanov V.K. Age-related structural features of liver and thyroid gland of laboratory rats. Vestnik KazNMU [Bulletin of KazNMU]. 2015; (3): 220–2. (in Russian).
19. Andreeva S.D. Cytokine status and destructive lesions of the liver in experimental acute pancreatitis. Ippologiya i veterinariya [Hippology and Veterinary]. 2020; (3): 69–73 (in Russian)
20. Conlon I., Raff M. Size control in animal development. Cell. 1999; 96 (2): 235–44. DOI: https://doi.org/10.1016/s0092-8674(00)80563-2
21. Sasaki K., Sonoda Y. Histometrical and three-dimensional analyses of liver hematopoiesis in the mouse embryo. Arch Histol Cytol. 2000; 63 (2): 137–46. DOI: https://doi.org/10.1679/aohc.63.137
22. Guo Y., Zhang X., Huang J., Zeng Y., Liu W., Geng C., et al. Relationships between hematopoiesis and hepatogenesis in the midtrimester fetal liver characterized by dynamic transcriptomic and proteomic profiles. PLoS One. 2009; 4 (10): e7641. DOI: https://doi.org/10.1371/journal.pone.0007641
23. Babaeva A.G. Regeneration: facts and perspectives. Moscow: RAMS Publishing House, 2009: 336 p. ISBN 978-5-7901-0085-7. (in Russian)
24. Timchenko N.A. Aging and liver regeneration. Trends Endocrinol Metab. 2009; 20 (4): 171–6. DOI: https://doi.org/10.1016/j.tem.2009.01.005
25. Baygil’din S.S., Karimov D.O., Khusnutdinova N.Yu., Smolyankin D.A., Repina E.F. Rat liver morphology after 72 hours of exposure to carbon. Meditsina truda i promyshlennaya ekologiya [Occupational Medicine and Industrial Ecology]. 2018; (4); 143–9. (in Russian)
26. Rui L. Energy metabolism in the liver. Comp Physiol. 2014; 4 (1): 177–97. DOI: https://doi.org/10.1002/cphy.c130024