MORPHOLOGICAL MONITORING OF EXPERIMENTAL LIVER FIBROSIS IN RATS
Abstract
Background. Though thioacetamide (TAA)-induced liver fibrosis (LF) is recognized as a classical model of toxic liver damage, there is no literature data on the description of its successive stages of histological and ultrastructural changes in various cell populations involved in fibrosis. Objective. To conduct morphological monitoring of fibrosis formation in the liver of rats using the TAA model of LF based on histological and ultrastructural changes in hepatocytes and perisinusoidal lipocytes (HSC). Material and methods. The experiment was carried out on 18 sexually mature male rats. LF was modeled by intraperitoneal injection of 2% TAA solution at a dose of 10 ml / kg every other day. Light microscopy of semi-thin sections of the liver was performed, as well as electron microscopy of ultrathin sections. Results. The study of semi-thin sections of rat liver tissue from the control group showed a normal architecture of the parenchyma, a large number of HSCs containing large lipid droplets ("resting" phenotype), a very small amount of cytoplasmic matrix poor in membrane organelles. In the animals that were receiving TAA for 4 weeks, a mesenchymalepithelial transition of HSCs from the "resting" type to a fibrogenic state (fibrogenic phenotype) was recorded, that was accompanied by a gradual decrease in the number of retinol-containing drops and the appearance of fibroblastlike cells (FLC) in HSCs. In the animals, that were receiving TAA for 12 weeks, the pool of fibrogenic cells in the liver increased, a mesothelial-mesenchymal transition occurred, characterized by the mesothelial cell migration deeper into the parenchyma and their acquisition of a mesenchymal phenotype. Lipid containing activated FLC were also found in fibrous tissue around the central vein. Foci of hepatic tissue destruction caused by necrosis and apoptosis of hepatocytes were much more common. Conclusions. Administration of TAA induces liver fibrosis while histological and ultrastructural monitoring of the state of hepatocytes and HSCs allows to monitor all stages of fibrosis, clarifying the mechanisms of damage to intracellular organelles and variants of hepatocyte death. This model of LF in rats can be used to test new antifibrotic drugs.
References
Delire B, Stärkel P, Leclercq I. Animal Models for Fibrotic Liver Diseases: What We Have, What We Need, and What Is under Development. J Clin Transl Hepatol. 2015;3(1):53-66. https://doi.org/10.14218/JCTH.2014.00035.
Starkel P, Leclercq IA. Animal models for the study of hepatic fibrosis. Best Pract Res Clin Gastroenterol. 2011;25(2):319-33. https://doi.org/10.1016/j.bpg.2011.02.004.
Hoffmann C, Djerir NEH, Danckaert A, Fernandes J, Roux P, Charrueau C, Lachagès AM, Charlotte F, Brocheriou I, Clément K, Aron-Wisnewsky J, Foufelle F, Ratziu V, Hainque B, Bonnefont-Rousselot D, Bigey P, Escriou V. Hepatic stellate cell hypertrophy is associated with metabolic liver fibrosis. Sci Rep. 2020;10(1):3850. https://doi.org/10.1038/s41598-020-60615-0.
Porter WR, Gudzinowicz MJ, Neal RA. Thioacetamideinduced hepatic necrosis. II. Pharmacokinetics of thioacetamide and thioacetamide-S-oxide in the rat. J Pharmacol Exp Ther. 1979;208(3):386-91.
Gilmore TD, Wolenski FS. NF-κB: where did it come from and why? Immunol Rev. 2012;246(1):14-35. https://doi.org/10.1111/j.1600-065X.2012.01096.x.
Akhtar T, Sheikh N. An overview of thioacetamide-induced hepatotoxicity. Toxin Reviews. 2013;32(3):43-46. https://doi.org/10.3109/15569543.2013.805144.
Tsyrkunov V, Andreev V, Kravchuk R, Kondratovich I. Ito stellate cells (hepatic stellate cells) in diagnosis of liver fibrosis. Gastroenterol Hepatol Open Access. 2019;10(4):213-219. https://doi.org/10.15406/ghoa.2019.10.00384.
Novogrodskaya Ya, Ostrovskaya O, Kravchuk R, Doroshenko Ye, Huliai I, Aleschyk A, Shelesnaya S, Kurbat M. Sposob modelirovanija jeksperimentalnogo tioacetamidnogo porazhenija pecheni u krys [The method of modelling of experimental thioacetamide liver damage in rats]. Gepatologija i gastrojenterologija [Hepatology and Gastroenterology]. 2020;4(1):90-95. https://doi.org/10.25298/2616-5546-2020-4-1-90-95. (Russian).
Sato T, Takagi I. An electron microscopic study of specimens fixed for longer periods in phosphate buffered formalin. J Electron Microsc. 1982;31(4):423-8.
Lee YH, Son JY, Kim KS, Park YJ, Kim HR, Park JH, Kim KB, Lee KY, Kang KW, Kim IS, Kacew S, Lee BM, Kim HS. Estrogen Deficiency Potentiates Thioacetamide-Induced Hepatic Fibrosis in Sprague-Dawley Rats. Int J Mol Sci. 2019;20(15):3709. https://doi.org/10.3390/ijms20153709.
Liedtke C, Luedde T, Sauerbruch T, Scholten D, Streetz K, Tacke F, Tolba R, Trautwein C, Trebicka J, Weiskirchen R. Experimental liver fibrosis research: update on animal models, legal issues and translational aspects. Fibrogenesis Tissue Repair. 2013;6(1):19. https://doi.org/10.1186/1755-1536-6-19.
An P, Wei LL, Zhao S, Sverdlov DY, Vaid KA, Miyamoto M, Kuramitsu K, Lai M, Popov YV. Hepatocyte mitochondria-derived danger signals directly activate hepatic stellate cells and drive progression of liver fibrosis. Nat Commun. 2020;11(1):2362. https://doi.org/10.1038/s41467-020-16092-0.
Tsyrkunov V, Chernyak S, Prokopchik N, Andreev V, Shulika V. Vlijanie bakterialnogo lipopolisaharida – pirogenala na regress fibroza v pecheni pri hronicheskom gepatite C [Impact of bacterial lipopolysaccharide – pyrogenal on a regression of liver fibrosis in chronic hepatitis C]. Recept [Recipe]. 2015;6:45-53. (Russian).
1.png)
