Immunohistochemical analysis of microglial changes in the experimental acute hepatic encephalopathy
DOI:
https://doi.org/10.14739/2310-1237.2021.1.227642Keywords:
acute hepatic encephalopathy, microglial, phagocytosis, CD68Abstract
Hepatic encephalopathy (HE) is a syndrome of impaired brain function in patients with advanced liver failure and it manifests in form of psychometric tests alterations up to decreased consciousness and coma. The current knowledge about HE mainly focused on the theory of ammonia neurotoxicity and neuroinflammation. Microglia being resident innate immune cells of the brain when activated are responsible for the neuroinflammatory reactions.
The aim – immunohistochemical study of the microglial changes in different rat brain regions in conditions of experimental acute HE (AHE).
Materials and methods. We used acetaminophen induced liver failure model in Wistar rats. Four from 10 animals that survived up to 24 h after acetaminophen injection constituted “compensated group”; 6 animals which died within 24 h – “decompensated group”. Microglial reactive changes were analysed by the evaluation of the relative area (S rel., %) of CD68+ expression in the brain cells not associated with meninges and vessels, as well as the changing in shape and number of these cells.
Results. Acetaminophen-induced AHE in rats was characterized by the regional- and time-dependent dynamic increase in CD68 expression level in the rat brain in form of significant (relatively to control) increase of CD68+ S rel. in brain cells and the number of such cells. The medians of CD68+ S rel. and their numbers in significantly changed regions of non-survived rats were, respectively: subcortical white matter – 0.24 (0.20; 0.26) and 11.00 (8.00; 13.00); thalamus – 0.13 (0.90; 0.18) and 6.00 (3.00; 7.00); caudate/putamen – 0.13 (0.12; 0.18) and 7.00 (4.00; 11.00) – all indicators were statistically significant compared to control. In the survived animals, indicators were, respectively: subcortical white matter – 0.24 (0.16; 0,26) and 10.00 (8.00; 12.00); caudate/putamen – 0.12 (0.10; 0.15) and 6.00 (4.00; 10.00) – the differences were significant compared to control.
Conclusions. The highest and significant indicators were revealed at 24 h (compared to earlier time points) of the experiment in the white matter, thalamus and caudate/putamen. This fact reflects time-dependent dynamic boosting of reactive changes in microglia and presumably may indicate the regions of the most active neuroinflammatory response within the brain parenchyma in the conditions of AHE. The appearing of a small percentage of cells with amoeboid transformation among CD68+-cells may mean partial functional insufficiency of such cells due to probable suppressive impact of ammonia or other influencing factors, as well as insignificance of the material that needs to be phagocytosed under established conditions.
References
Ferenci, P. (2017). Hepatic encephalopathy. Gastroenterology report, 5(2), 138-147. https://doi.org/10.1093/gastro/gox013
Flamm, S. L. (2018). Complications of Cirrhosis in Primary Care: Recognition and Management of Hepatic Encephalopathy. The American journal of the medical sciences, 356(3), 296-303. https://doi.org/10.1016/j.amjms.2018.06.008
Zemtsova, I., Görg, B., Keitel, V., Bidmon, H. J., Schrör, K., & Häussinger, D. (2011). Microglia activation in hepatic encephalopathy in rats and humans. Hepatology, 54(1), 204-215. https://doi.org/10.1002/hep.24326
Jayakumar, A. R., Rama Rao, K. V., & Norenberg, M. D. (2015). Neuroinflammation in hepatic encephalopathy: mechanistic aspects. Journal of clinical and experimental hepatology, 5(Suppl 1), S21-S28. https://doi.org/10.1016/j.jceh.2014.07.006
Rao, K. V., Brahmbhatt, M., & Norenberg, M. D. (2013). Microglia contribute to ammonia-induced astrocyte swelling in culture. Metabolic brain disease, 28(2), 139-143. https://doi.org/10.1007/s11011-012-9339-1
Balzano, T., Dadsetan, S., Forteza, J., Cabrera-Pastor, A., Taoro-Gonzalez, L., Malaguarnera, M., Gil-Perotin, S., Cubas-Nuñez, L., Casanova, B., Castro-Quintas, A., Ponce-Mora, A., Arenas, Y. M., Leone, P., Erceg, S., Llansola, M., & Felipo, V. (2020). Chronic hyperammonemia induces peripheral inflammation that leads to cognitive impairment in rats: Reversed by anti-TNF-α treatment. Journal of hepatology, 73(3), 582-592. https://doi.org/10.1016/j.jhep.2019.01.008
Rangroo Thrane, V., Thrane, A. S., Chang, J., Alleluia, V., Nagelhus, E. A., & Nedergaard, M. (2012). Real-time analysis of microglial activation and motility in hepatic and hyperammonemic encephalopathy. Neuroscience, 220, 247-255. https://doi.org/10.1016/j.neuroscience.2012.06.022
Karababa, A., Groos-Sahr, K., Albrecht, U., Keitel, V., Shafigullina, A., Görg, B., & Häussinger, D. (2017). Ammonia Attenuates LPS-Induced Upregulation of Pro-Inflammatory Cytokine mRNA in Co-Cultured Astrocytes and Microglia. Neurochemical research, 42(3), 737-749. https://doi.org/10.1007/s11064-016-2060-4
Stankov, A., Belakaposka-Srpanova, V., Bitoljanu, N., Cakar, L., Cakar, Z., & Rosoklija, G. (2015). Visualisation of Microglia with the use of Immunohistochemical Double Staining Method for CD-68 and Iba-1 of Cerebral Tissue Samples in Cases of Brain Contusions. Prilozi, 36(2), 141-145. https://doi.org/10.1515/prilozi-2015-0062
Masuda, T., Amann, L., Sankowski, R., Staszewski, O., Lenz, M., D Errico, P., Snaidero, N., Costa Jordão, M. J., Böttcher, C., Kierdorf, K., Jung, S., Priller, J., Misgeld, T., Vlachos, A., Meyer-Luehmann, M., Knobeloch, K. P., & Prinz, M. (2020). Novel Hexb-based tools for studying microglia in the CNS. Nature immunology, 21(7), 802-815. https://doi.org/10.1038/s41590-020-0707-4
De Biase, L. M., & Bonci, A. (2019). Region-Specific Phenotypes of Microglia: The Role of Local Regulatory Cues. The Neuroscientist, 25(4), 314-333. https://doi.org/10.1177/1073858418800996
Mitchell, R. A., Rathi, S., Dahiya, M., Zhu, J., Hussaini, T., & Yoshida, E. M. (2020). Public awareness of acetaminophen and risks of drug induced liver injury: Results of a large outpatient clinic survey. PloS one, 15(3), e0229070. https://doi.org/10.1371/journal.pone.0229070
McGill, M. R., Williams, C. D., Xie, Y., Ramachandran, A., & Jaeschke, H. (2012). Acetaminophen-induced liver injury in rats and mice: comparison of protein adducts, mitochondrial dysfunction, and oxidative stress in the mechanism of toxicity. Toxicology and applied pharmacology, 264(3), 387-394. https://doi.org/10.1016/j.taap.2012.08.015
Mossanen, J. C., & Tacke, F. (2015). Acetaminophen-induced acute liver injury in mice. Laboratory animals, 49(1 Suppl), 30-36. https://doi.org/10.1177/0023677215570992
Shulyatnikova, T., & Shavrin, V. (2021). Mobilisation and redistribution of multivesicular bodies to the endfeet of reactive astrocytes in acute endogenous toxic encephalopathies. Brain research, 1751, 147174. https://doi.org/10.1016/j.brainres.2020.147174
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