Predictors of liver fibrosis during ligation of the common bile duct in rats

V. M.  Ratchyk; T. G.  Turytska

doi:10.15421/0224106

V. M. Ratchyk Oles Honchar Dnipro National University
T. G. Turytska Dnipro State Medical University

Keywords: obstructive jaundice; experimental cholestasis; cytokines; fibrosis; liver cirrhosis; purulent cholangitis

Abstract

Determination of predictors of liver fibrosis during ligation of the common bile duct in experimental animals demonstrates the dynamics of the development of biochemical, immunological, and morphological markers of cholestatic disease, which is subsequently accompanied by the formation of liver cirrhosis. An increase in the degree of obstructive jaundice caused pronounced negative changes in the clinical picture of the disease and morphostructural changes of the liver, which, depending on the time of obstruction, became irreversible. In the conditions of a chronic experiment on rats with the aim of identifying patterns of development of obstructive jaundice, assessing the processes of liver damage and developing treatment directions, modeling of obstructive jaundice was performed with further study of the clinical picture and behavioral reactions, biochemical, immunological, morphological data depending on the duration of obstruction of the biliary tract. During the study, three stages of disease development were established: initial (up to 7 days), compensatory (8–15 days) and decompensation stage (from 16 days after ligation of the common bile duct). Hyperbilirubinemia and accumulation of bile acids, an increase in the content of medium-weight molecules with the activation of lipoperoxidation and endotoxemia occurred against the background of strengthening the processes of connective tissue organization. The assessment of the effect of cytokines made it possible to establish a reliable negative correlation between IL-6, TNF-α and IL-10, which led to the strengthening of cytotoxic reactions. Against the background of an imbalance of the cytokine chain under the conditions of the experiment, obstruction of the biliary tract from the first days of ligation was accompanied by the expression of the profibrogenic cytokine TGF-β1 with the activity of fibrotic processes in the liver parenchyma, while a reliable relationship was established between an increase in the concentration of TGF-β1 and fibrosis (r = 0.445; P = 0.041). When assessing the histostructure of rat livers, after 20 days, developing cirrhosis was detected in 25.0% of the animals. Micronodular cirrhosis with loss of the beam structure of the liver parenchyma and fibrosis of the lobes was reproduced in 33.3% of the animals. Against the background of cholestatic hepatitis, secondary obstructive biliary cirrhosis of the liver was reproduced in 58.3% of the animals. Ligation of the common bile duct in rats causes inflammatory, necrobiotic, dystrophic changes in the liver, which, against the background of increasing endogenous intoxication, increased lipoperoxidation and cholestatic phenomena, stimulate a cascade of processes of excessive organization of connective tissue, the final effect of which is the formation of liver cirrhosis. Prospects for further research are the development of processes of influence on the mechanisms of liver regeneration and slowing down of fibrotic reactions. Additional studies are needed to determine changes in liver tissue in the posticteric period and factors affecting the healing of cholestatic liver damage and regeneration of hepatocytes.

References

Akkız, H., Gieseler, R. K., & Canbay, A. (2024). Liver fibrosis: From basic science towards clinical progress, focusing on the central role of hepatic stellate cells. International Journal of Molecular Sciences, 25(14), 7873.

Arjmand, A., Tsipouras, M. G., Tzallas, A. T., Forlano, R., Manousou, P., & Giannakeas, N. (2020). Quantification of liver fibrosis – a comparative study. Applied Sciences, 10(2), 447.

Boyer, J. L. (2007). New perspectives for the treatment of cholestasis: Lessons from basic science applied clinically. Journal of Hepatology, 46(3), 365–371.

Bulte, J. W., Schmieder, A. H., Keupp, J., Caruthers, S. D., Wickline, S. A., & Lanza, G. M. (2014). MR cholangiography demonstrates unsuspected rapid biliary clearance of nanoparticles in rodents: Implications for clinical translation. Nanomedicine: Nanotechnology, Biology, and Medicine, 10(7), 1385–1388.

Daneze, E. R., Terra, G. A., Júnior, J. A. T., Campos, A. G., da Silva, A. A., & Terra, S. A. (2011). Comparative study between ligature with thread or metallic clamping by means of laparoscopy with the purpose of experimental biliary obstruction in swines. Journal of Acta Cirurgica Brasileira, 26(2), 31–37.

Donato, M. T., Gallego-Ferrer, G., & Tolosa, L. (2022). In vitro models for studying chronic drug-induced liver injury. International Journal of Molecular Sciences, 23(19), 11428.

Geh, D., Manas, D. M., & Reeves, H. L. (2021). Hepatocellular carcinoma in non-alcoholic fatty liver disease-a review of an emerging challenge facing clinicians. Hepatobiliary Surgery and Nutrition, 10(1), 59–75.

Ginès, P., Krag, A., Abraldes, J. G., Solà, E., Fabrellas, N., & Kamath, P. S. (2021). Liver cirrhosis. Lancet, 398(10308), 1359–1376.

Gong, Y. H., & Li, W. (2008). The different effects of internal and external biliary drainages on the blood levels of endotoxin, interleukin-2 and interleukin-6 in rats with obstructive jaundice. Weichangbingxue He Ganbingxue Zazhi, 4, 329–331.

Ishiwatari, H., Sato, J., & Sakamoto, H. (2024). Endoscopic biliary drainage for biliary stricture. Nihon Shokakibyo Gakkai Zasshi, 121(4), 275–286.

Jorge, G. D. L., Leonardi, L. S., Boin I. F. S. F., Silva, J. O. C., & Escanhoela C. A. F. (2001). A new method for the experimental induction of secundary biliary cirrhosis in wistar rats. Journal of Acta Cirurgica Brasileira, 16(2), 75–81.

Kiani, A. K., Pheby, D., Henehan, G., Brown, R., Sieving, P., Sykora, P., Marks, R., Falsini, B., Capodicasa, N., Miertus, S., Lorusso, L., Dondossola, D., Tartaglia, G. M., Ergoren, M. C., Dundar, M., Michelini, S., Malacarne, D., Bonetti, G., Dautaj, A., Donato, K., Medori, M. C., Beccari, T., Samaja, M., Connelly, S. T., Martin, D., Morresi, A., Bacu, A., Herbst, K. L., Kapustin, M., Stuppia, L., Lumer, L., Farronato, G., Bertelli, M., & International Bioethics Study Group (2022). Ethical considerations regarding animal experimentation. Journal of Preventive Medicine and Hygiene, 63(2 Suppl. 3), e255–e266.

Krzemień, G., Szmigielska, A., Turczyn, A., & Pańczyk-Tomaszewska, M. (2016). Urine interleukin-6, interleukin-8 and transforming growth factor β1 in infants with urinary tract infection and asymptomatic bacteriuria. Central-European Journal of Immunology, 41(3), 260–267.

Kutovyi, O. B., Rodynska, H. O., & Balyk, D. V. (2018). Dosvid likuvannia khvorykh iz syndromom tiazhkoi mekhanichnoi zhovtianytsi dobroiakisnoi etiolohii [Experience of treating patients with the syndrome of severe mechanical jaundice of benign etiology]. Ukrainskyi Zhurnal Khirurhii, 37, 36–40 (in Ukrainian).

Lee, H., Yu, D. M., Bahn, M. S., Kwon, Y. J., Um, M. J., Yoon, S. Y., Kim, K. T., Lee, M. W., Jo, S. J., Lee, S., Koo, S. H., Jung, K. H., Lee, J. S., & Ko, Y. G. (2022). Hepatocyte-specific Prominin-1 protects against liver injury-induced fibrosis by stabilizing SMAD7. Experimental and Molecular Medicine, 54(8), 1277–1289.

Liu, J. J., Sun, Y. M., Xu, Y., Mei, H. W., Guo, W., & Li, Z. L. (2023). Pathophysiological consequences and treatment strategy of obstructive jaundice. World Journal of Gastrointestinal Surgery, 15(7), 1262–1276.

Long, Y., Dong, X., Yuan, Y., Huang, J., Song, J., Sun, Y., Lu, Z., Yang, L., & Yu, W. (2015). Metabolomics changes in a rat model of obstructive jaundice: Mapping to metabolism of amino acids, carbohydrates and lipids as well as oxidative stress. Journal of Clinical Biochemistry and Nutrition, 57(1), 50–59.

Lv, Y., Yue, J., Gong, X., Han, X., Wu, H., Deng, J., & Li, Y. (2018). Spontaneous remission of obstructive jaundice in rats: selection of experimental models. Experimental and Therapeutic Medicine, 15(6), 5295–5301.

Ma, J., Zhao, Q., Chen, M., Wang, W., He, B., Jiang, Y., & Li, Y. (2022). MicroRNA-122 inhibits hepatic stellate cell proliferation and activation in vitro and represses carbon tetrachloride-induced liver cirrhosis in mice. Annals of Hepatology, 27(4), 100700.

Moraes, P. A. D., Tannuri, A. C. A., Rios, L. M., Paes, V. R., Gonçalves, J. O., Serafini, S., & Tannuri, U. (2020). Sepsis and cirrhosis in growing animals: description of a new experimental model and its pathological and immunological reliability. Clinics, 75, e1858.

Novo, E., Bocca, C., Foglia, B., Protopapa, F., Maggiora, M., Parola, M., & Cannito, S. (2020). Liver fibrogenesis: Un update on established and emerging basic concepts. Аrchives of Biochemistry and Biophysics, 689, 108445.

Odisseos, C., Ioannidis, O., Chatzakis, C., Symeonidis, S., Bitsianis, S., Christidis, P., Loutzidou, L., Mantzoros, I., Kotidis, E., Pramateftakis, M. G., Angelopoulos, S., & Tsalis, K. (2020). The effect of hepatic ischemia in the liver of rats with obstructive jaundice. Annali Italiani di Chirurgia, 91, 334–344.

Oguz, S., Salt, O., Ibis, A.C., Gurcan, S., Albayrak, D., Yalta, T., Sagiroglu, T., & Erenoglu, C. (2018). Combined effectiveness of honey and immunonutrition on bacterial translocation secondary to obstructive jaundice in rats: Experimental study. Medical Science Monitor, 24, 3374–3381.

Ozozan, O. V., Dinc, T., Vural, V., Ozogul, C., Ozmen, M. M., & Coskun, F. (2020). An electron microscopy study of liver and kidney damage in an experimental model of obstructive jaundice. Annali Italiani di Chirurgia, 91, 122–130.

Paulusma, C. C., Lamers, W. H., Broer, S., & van de Graaf, S. F. J. (2022). Amino acid metabolism, transport and signalling in the liver revisited. Biochemical Pharmacology, 201, 115074.

Pavlidis, E. T., & Pavlidis, T. E. (2018). Pathophysiological consequences of obstructive jaundice and perioperative management. Hepatobiliary and Pancreatic Diseases International, 17(1), 17–21.

Pellicoro, A., Ramachandran, P., Iredale, J. P., & Fallowfield, J. A. (2014). Liver fibrosis and repair: Immune regulation of wound healing in a solid organ. Nature Reviews, Immunology, 14(3), 181–194.

Petrie, A., & Sabin, C. (2020). Medical statistics at a glance. 2th ed. Wiley-Blackwell, Oxford.

Pieters, A., Gijbels, E., Cogliati, B., Annaert, P., Devisscher, L., & Vinken, M. (2021). Biomarkers of cholestasis. Biomarkers in Medicine, 15(6), 437–454.

Roy, S. K., & Lambert, A. (2017). Obstructive jaundice: A clinical review for the UK armed forces. Journal of the Royal Naval Medical Service, 103(1), 44–48.

Salas-Silva, S., Simoni-Nieves, A., Chávez-Rodríguez, L., Gutiérrez-Ruiz, M. C., Bucio, L., & Quiroz, L. E. G. (2021). Mechanism of cholangiocellular damage and repair during cholestasis. Annals of Hepatology, 26, 100530.

Salas-Silva, S., Simoni-Nieves, A., Lopez-Ramirez, J., Bucio, L., Gómez-Quiroz, L. E., Gutiérrez-Ruiz, M. C., & Roma, M. G. (2019). Cholangiocyte death in ductopenic cholestatic cholangiopathies: Mechanistic basis and emerging therapeutic strategies. Life Sciences, 218, 324–339.

Schuppan, D., & Afdhal, N. H. (2008). Liver cirrhosis. Lancet, 371(9615), 838–851.

Terai, S., & Sakamaki, A. (2021). Problems and future questions in the clinical practice of liver cirrhosis. Nihon Shokakibyo Gakkai Zasshi, 118(1), 41–45.

Terai, S., & Tsuchiya, A. (2017). Status of and candidates for cell therapy in liver cirrhosis: Overcoming the “point of no return” in advanced liver cirrhosis. Journal of Gastroenterology, 52(2), 129–140.

Thakkar, N., Slizgi, J. R., & Brouwer, K. L. R. (2017). Effect of liver disease on hepatic transporter expression and function. Journal of Pharmaceutical Sciences, 106(9), 2282–2294.

Turk, O., Badak, B., Ates, E., Dundar, E., & Sutken, E. (2016). The role of growth factors on hepatic damage in rats with obstructive jaundice. SpringerPlus, 5(1), 1274.

Van Campenhout, S., Van Vlierberghe, H., & Devisscher, L. (2019). Common bile duct ligation as model for secondary biliary cirrhosis. Methods in Molecular Biology, 1981, 237–247.

Wang, X. L., Yang, M., & Wang, Y. (2024). Roles of transforming growth factor-β signaling in liver disease. World Journal of Hepatology, 16(7), 973–979.

Yang, J. J., Yang, Y., Zhang, C., Li, J., & Yang, Y. (2020). Epigenetic silencing of LncRNA ANRIL enhances liver fibrosis and HSC activation through activating AMPK pathway. Journal of Cellular and Molecular Medicine, 24(4), 2677–2687.

Yorganci, K., Baykal, A., Kologlu, M., Saribaş, Z., Hascelik, G., & Sayek, I. (2004). Endotoxin challenge causes a proinflammatory state in obstructive jaundice. Journal of Investigative Surgery, 17(3), 119–126.

Zapadnjuk, I. P., Zapadnjuk, E. A., Zaharija, E. A., & Zapadnjuk, B. V. (1983). Laboratornye zhivotnye: Razvedenie, soderzhanie, ispol’zovanie v eksperimente [Laboratory animals: Breeding, content, use in experiment]. Vishha Shkola, Kiev (in Ukrainian).

Zhou, P. H., Yao, L. Q., Zhang, Y. Q., Gao, W. D., He, G. J., Xu, M. D., Wang, P., & Qin, X. Y. (2003). Endoscopic biliary drainage for biliary obstruction. Hepatobiliary and Pancreatic Diseases International, 2(4), 598–601.