Regenerative response of hepatocytes in the course of diffuse toxic damage

Z.A. Shafigullina¹, I.F. Gette¹, I.G. Danilova¹,²

¹Institute of Immunology and Physiology Ural Branch of RAS, Yekaterinburg, Russian Federation;

²Institute of Medical Cellular Technologies, Yekaterinburg, Russian Federation

Abstract. The study of liver regeneration in the course of its toxic damage in order to prolong the reserves of hepatocyte functioning is an important problem. Objective: to determine the features of the regenerative strategy of hepatocytes in the course of diffuse toxic damage. Materials and methods. Toxic liver damage of male Wistar rats was caused by a single intraperitoneal injection of an oil solution of carbon tetrachloride (CCl₄) at a dose of 50 mg/100 g. Animals were carried out from the experiment on days 3 and 14. The study used biochemical, immunohistochemical research methods, light and confocal microscopy, and statistical analysis. Results. It was found that in response to the action of hepatotropic toxin on day 3, along with destructive processes (structural disorders, increased apoptosis, an imbalance in biochemical parameters), there is an increase in the number of proliferating and binuclear hepatocytes, along with the number of HSP70⁺ hepatocytes. By 14 days after administration of the toxicant, recovery processes are implemented mainly due to intracellular regeneration, as evidenced by an increase in the number of binuclear hepatocytes. Biochemical parameters are normalized, but with reduced HSP60 expression in liver parenchyma cells, apoptotic cell death increases. Conclusion. Features of the regenerative strategy of hepatocytes in the early stages after exposure to hepatotropic venom are manifested in an increase in the mitotic activity of parenchymal cells and simultaneous activation of intracellular recovery processes, accompanied by an increase in binuclear cells and an increase in the metabolic activity of preserved hepatocytes. The dominant type of regeneration in the long term, on the 14th day after intraperitoneal administration of CCl₄, is intracellular regeneration, due to which biochemical parameters return to normal. The regenerative response of hepatocytes is also expressed in changes in the production of heat shock proteins 60 and 70 kDa. An increase in the number of hepatocytes containing Hsp70 granules in the early stages of toxicant exposure can serve as a deterrent to hepatocyte apoptosis, while a decrease in HSP60⁺ hepatocytes, on the contrary, serves as an apoptotic stimulus.

Keywords: regeneration, hepatocyte, liver, apoptosis, heat shock proteins, toxic damage

There is no conflict of interest.

Contact details of the corresponding author:

Zlata A. Shafigullina

zlata_pyankova@mail.ru

Received 14.12.2020

For citation:

Shafigullina Z.A., Gette I.F., Danilova I.G. Regenerative response of hepatocytes in the course of diffuse toxic damage. http://vestnikural.ru/article/1153 [Online] Vestn. Ural. Med. Akad. Nauki. = Journal of Ural Medical Academic Science. 2020, Vol. 17, no. 4, pp. 313–322. DOI: 10.22138/2500-0918-2020-17-4-313-322 (In Russ)

Figure 1. The structure of liver of experimental animals. Hematoxylin-eosin staining, mag. ×400.

Note: 1 — intact group; 2 — CCl₄ 3 days, 3 и 4 — CCl₄ 14 days; A — plethora of central veins and veins of portal tracts, B — focal infiltration, C — hepatocyte necrosis.

Figure 2. The degree of apoptosis and quantitative changes in hepatocytes with the content of heat shock protein granules (HSP70 and HSP60)

Note:* – statistically significant differences (p <0.05) with the group of intact animals, # – statistically significant differences (p <0.05) with the group of CCl₄ 3 days.

Figure 3. Indicators of liver regeneration in normal conditions and in the course of diffuse toxic damage

Note:* – statistically significant differences (p<0.05) with the group of intact animals, # – statistically significant differences (p <0.05) with the group of CCl₄ 3 days.

Table 1

Biochemical parameters of experimental animals in the dynamics of diffuse toxic damage

Note:* – statistically significant differences (p<0.05) with the group of intact animals, # – statistically significant differences (p<0.05) with the group of CCl₄ 3 days.

REFERENCES

1. Forbes S., Newsome P. Liver regeneration – mechanisms and models to clinical application. Nat Rev Gastroenterol Hepatol. 2016, 13: 473–485. doi: 10.1038/nrgastro.2016.97.
2. Gu X., Manautou J. E. Molecular mechanisms underlying chemical liver injury. Expert Rev Mol Med. 2012, 14: e4. doi:10.1017/S1462399411002110.
3. Khan A. A., Alzohairy M. A. Hepatoprotective effects of camel milk against CCl₄ induced hepatotoxicity in rats. Asian Journal of Biochemistry. 2011, 6: 171–180.  doi:  10.3923/ajb.2011.171.180.
4. Mahmood K., Jadoon S., Mahmood Q., Irshad M., Hussain J. Synergistic effects of toxic elements on heat shock proteins. BioMed research international. 2014: 564136. doi: 10.1155/2014/564136.
5. Walter S., Buchner J. Molecular chaperones-cellular machines for protein folding. Angew Chem Int Ed Engl. 2002. 41(7): 1098–113.
6. Shi J., Aisaki K., Ikawa Y., Wake K. Evidence of hepatocyte apoptosis in rat liver after the administration of carbon tetrachloride. Am J Pathol. 1998. 153: 515–525.
7. Guidelines for conducting preclinical studies of drugs. Part one. M .: Grif i K, 2012.944 p. (In Russ.)
8. Zahedi K., Barone S. L., Xu J., Steinbergs N., Schuster R., Lentsch A. B. et al. Hepatocyte-specific ablation of spermine/spermidine-N1-acetyltransferase gene reduces the severity of CCl₄-induced acute liver injury. Am J Physiol Gastrointest Liver Physiol. 2012. 303: 546–560.
9. Taganovich AD et al. Biological chemistry: 2nd ed., Revised. Minsk: Higher School, 2016.671 p. (In Russ.)
10. Kaletina N. I. ed. Toxicological chemistry. Metabolism and analysis of toxicants: a tutorial. M.: GEOTAR-Media, 2008.1016 p. (In Russ.)
11. Kurashova N. A., Madaeva I. M., Kolesnikova L. I. Expression of HSP70 Heat-Shock Proteins under Oxidative Stress. Adv Gerontol. 2020. 10: 20–25. doi: 10.1134/S2079057020010099.

Authors

Zlata A. Shafigullina

Institute of Immunology and Physiology Ural Branch of RAS

Junior researcher Laboratory of Morphology and Biochemistry

106, Pervomayskay street, Yekaterinburg, Russian Federation, 620049

zlata_pyankova@mail.ru

Irina F. Gette

Institute of Immunology and Physiology Ural Branch of RAS

Cand. Sci. (Bio.), Senior researcher Laboratory of Morphology and Biochemistry

106, Pervomayskay street, Yekaterinburg, Russian Federation, 620049

i.goette@yandex.ru

Irina G. Danilova

Institute of Immunology and Physiology Ural Branch of RAS

Dr. Sci. (Bio.), docent, Head of the Laboratory of Morphology and Biochemistry

Institute of Medical Cellular Technologies

Chief Researcher

22a Karl Marks street, Yekaterinburg, Russian Federation, 620026

ig-danilova@yandex.ru