Radiological study of lung tissue in rats with bleomycin-induced fibrosis

Yuliia Paramonova, Anatoliy Mazurkevich, Iurii Kharkevych, Taras Savchuk
Abstract

The increasing incidence of pulmonary fibrosis in animals caused by toxic exposure necessitates a detailed analysis of radiological changes in lung tissue. This is essential for assessing the progression of fibrosis and developing effective therapeutic approaches. In this context, the study aimed to utilise radiological methods to identify time-dependent changes in the lungs of rats subjected to a model of bleomycin-induced pulmonary fibrosis, contributing to the improvement of diagnostic techniques for this pathological condition. Pulmonary fibrosis was experimentally induced in laboratory rats by administering bleomycin. Radiological examinations were conducted on days 7, 14, 30, and 45 to monitor the condition of lung tissue and detect progressive changes. Particular attention was given to identifying characteristic radiological signs of fibrosis, such as lung tissue consolidation and reduced transparency. The findings revealed a progressive nature of interstitial lesions. It was established that as early as seven days after bleomycin administration, signs of inflammation appeared in the lungs, accompanied by reduced transparency of the pulmonary fields. By day 14, initial signs of fibrosis were observed, marked by increased lung tissue density and the presence of infiltrates. By day 30 of pathology progression, significant fibrosis development was evident, characterised by more pronounced linear inclusions and reduced transparency. By day 45, scar tissue formation occurred, leading to substantial structural deterioration of the lungs. The findings highlighted the importance of radiological examination as an informative method for diagnosing and monitoring the progression of pulmonary fibrosis. The radiological changes observed at different stages of fibrosis development enable a more detailed characterisation of the disease pathogenesis, which is crucial for evaluating the effectiveness of therapeutic interventions. Accordingly, the study results could be utilised to establish new diagnostic criteria and enhance the monitoring of patients with pulmonary fibrosis

Keywords

pulmonary fibrosis; radiographic imaging; interstitial lung disease; experimental model; laboratory rats; diagnosis

Suggested citation
Paramonova, Yu., Mazurkevich, A., Kharkevych, Iu., & Savchuk, T. (2024). Radiological study of lung tissue in rats with bleomycin-induced fibrosis. Ukrainian Journal of Veterinary Sciences, 15(4), 63-79. https://doi.org/10.31548/veterinary4.2024.63
References

[1] Abu Qubo, A., Capaccione, K.M., Bernstein, E.J., Padilla, M.L., & Salvatore, M.M. (2022). The role of radiology in progressive fibrosing interstitial lung disease. Frontiers in Medicine, 8, article number 679051. doi: 10.3389/fmed.2021.679051.

[2] Ayilya, B.L., Balde, A., Ramya, M., Benjakul, S., Kim, S., & Nazeer, R.A. (2023). Insights on the mechanism of bleomycin to induce lung injury and associated in vivo models: A review. International Immunopharmacology, 121, article number 110493. doi: 10.1016/j.intimp.2023.110493.

[3] Cony, F.G., Argenta, F.F., Heck, L.C., Moreira, L.F., Costa, F.V., Sonne, L., & Pavarini, S.P. (2019). Clinical and pathological aspects of idiopathic pulmonary fibrosis in cats. Pesquisa Veterinária Brasileira, 39(02), 134-141. doi: 10.1590/1678-5150-PVB-5942.

[4] Courtoy, G.E., Leclercq, I.A., Froidure, A., Schiano, G., Morelle, J., Devuyst, O., Huaux, F., & Bouzin, C. (2020). Digital image analysis of picrosirius red staining: A robust method for multiorgan fibrosis quantification and characterization. Biomolecules, 10(11), article number 1585. doi: 10.3390/biom10111585.

[5] Craven, A., et al. (2024). Clinical findings and outcome predictors for multinodular pulmonary fibrosis in horses: 46 cases (2009-2019). Journal of Veterinary Internal Medicine, 38(3), 18421857. doi: 10.1111/jvim.17084.

[6] Directive 2010/63/EU of the European Parliament and of the Council on the protection of animals used for scientific purposes (2010, September). Retrieved from https://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2010:276:0033:0079:en:PDF.

[7] Doğan, E., Okur, S., & Hayirli, A. (2022). Vertebral heart score and cardiothoracic ratio in Wistar rats. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 70(1), 43-48. doi: 10.33988/auvfd.905135/.

[8] Easton-Jones, C.A., Cissell, D.D., Mohr, F.C., Chigerwe, M., & Pusterla, N. (2020). Prognostic indicators and long-term survival in 14 horses with equine multinodular pulmonary fibrosis. Equine Veterinary Education, 32(11), 41-46. doi: 10.1111/eve.13204.

[9] Ferrazza, A.M., & Baldassarri, P. (2020). Presentation and diagnosis of interstitial lung diseases. In Thoracic radiology (pp. 105-125). Cham: Springer. doi: 10.1007/978-3-030-35765-8_9.

[10] Gassert, F.T., et al. (2021). X-ray dark-field chest imaging: Qualitative and quantitative results in healthy humans. Radiology, 301(2), 389-395. doi: 10.1148/radiol.2021210963.

[11] Gupta, N., Paryani, M., Patel, S., Bariya, A., Srivastava, A., Pathak, Y., & Butani, S. (2024). Therapeutic strategies for idiopathic pulmonary fibrosis – thriving present and promising tomorrow. The Journal of Clinical Pharmacology, 64(7), 779-798. doi: 10.1002/jcph.2408.

[12] He, A., He, L., Chen, T., Li, X., & Cao, C. (2024). Biomechanical properties and cellular responses in pulmonary fibrosis. Bioengineering, 11(8), article number 747. doi: 10.3390/bioengineering11080747.

[13] Hjerpe, A., Abd Own, S., & Dobra, K. (2020). Integrative approach to cytologic and molecular diagnosis of malignant pleural mesothelioma. Translational Lung Cancer Research, 9(3), 934943. doi: 10.21037/tlcr-2019-pps-10.

[14] Hoffman, T.W., van Es, H.W., Biesma, D.H., & Grutters, J.C. (2022). Potential interstitial lung abnormalities on chest X-rays prior to symptoms of idiopathic pulmonary fibrosis. BMC Pulmonary Medicine, 22, article number 329. doi: 10.1186/s12890-022-02122-8.

[15] Ishida, Y., Kuninaka, Y., Mukaida, N., & Kondo, T. (2023). Immune mechanisms of pulmonary fibrosis with bleomycin. International Journal of Molecular Sciences, 24(4), article number 3149. doi: 10.3390/ijms24043149.

[16] Keshavan, S., Bannuscher, A., Drasler, B., Barosova, H., Petri-Fink, A., & Rothen-Rutishauser, B. (2023). Comparing species-different responses in pulmonary fibrosis research: Current understanding of in vitro lung cell models and nanomaterials. European Journal of Pharmaceutical Sciences: Official Journal of the European Federation for Pharmaceutical Sciences, 183, article number 106387. doi: 10.1016/j.ejps.2023.106387.

[17] Kolb, P., Upagupta, C., Vierhout, M., Ayaub, E.A., Bellaye, P., Gauldie, J., Shimbori, C., Inman, M.D., Ask, K., & Kolb, M.R.J. (2020). The importance of interventional timing in the bleomycin model of pulmonary fibrosis. European Respiratory Journal, 55(6), article number 1901105. doi: 10.1183/13993003.01105-2019.

[18] Laurila, H.P., & Rajamäki, M.M. (2020). Update on canine idiopathic pulmonary fibrosis in West Highland white terriers. Veterinary Clinics of North America: Small Animal Practice, 50(2), 431-446. doi: 10.1016/j.cvsm.2019.11.004.

[19] Law of Ukraine No. 3447-IV “On the Protection of Animals from Cruelty”. (2006, February). Retrieved from https://zakon.rada.gov.ua/laws/show/3447-15#Text.

[20] Lee, K.S., Han, J., Wada, N., Hata, A., Lee, H.Y., Yi, C.A., Hino, T., Doyle, T.J., Franquet, T., & Hatabu, H. (2023). Imaging of pulmonary fibrosis: An update, from the AJR special series on imaging of fibrosis. American Journal of Roentgenology, 222(2), article number e2329119. doi: 10.2214/AJR.23.29119.

[21] Li, D., Liu, Z., Luo, L., Tian, S., & Zhao, J. (2022). Prediction of pulmonary fibrosis based on X-rays by deep neural network. Journal of Healthcare Engineering, 13, article number 3845008. doi: 10.1155/2022/3845008.

[22] Libório-Ramos, S., Barbosa-Matos, C., Fernandes, R., Borges-Pereira, C., & Costa, S. (2023). Іnterstitial macrophages lead early stages of bleomycin-induced lung fibrosis and induce fibroblasts activation. Cells, 12(3), article number 402. doi: 10.3390/cells12030402.

[23] Malinczak, C.A., Fonseca, W., Hrycaj, S.M., Morris, S.B., Rasky, A.J., Yagi, K., Wellik, D.M., Ziegler, S.F., Zemans, R.L., & Lukacs, N.W. (2024). Early-life pulmonary viral infection leads to long-term functional and lower airway structural changes in the lungs. American Journal of Physiology. Lung Cellular and Molecular Physiology, 326(3), 280-291. doi: 10.1152/ajplung.00300.2023.

[24] Mattoo, H.R., & Pillai, S. (2021). Idiopathic pulmonary fibrosis and systemic sclerosis: Pathogenic mechanisms and therapeutic interventions. Cellular and Molecular Life Sciences, 78(14), 5527-5542. doi: 10.1007/s00018-021-03874-y.

[25] Mazurkevych, A.Y., & Surtaieva, Y.V. (2023). Restoration of the morphofunctional state of rats lungs with experimental fibrosis through transplanted stem cells. Agricultural Science and Practice, 10(1), 12-23. doi: 10.15407/agrisp10.01.012.

[26] Mohr, W.L. (1988). Diffuse fibrosing alveolitis in three adult cattle. The Bovine Practitioner, 23, 156-158. doi: 10.21423/bovine-vol0no23p156-158.

[27] Ortiz-Zapater, E., et al. (2022). Epithelial coxsackievirus adenovirus receptor promotes house dust mite-induced lung inflammation. Nature Communications, 13, article number 6407. doi: 10.1038/s41467-022-33882-w.

[28] Persson, I.M., Håkansson, H.F., Örbom, A., Liu, J., von Wachenfeldt, K., & Olsson, L.E. (2020). Imaging biomarkers and pathobiological profiling in a rat model of drug-induced interstitial lung disease induced by bleomycin. Frontiers in Physiology, 11, article number 584. doi: 10.3389/fphys.2020.00584.

[29] Plumb, D. (2008). Plumb’s veterinary drug handbook (6th ed.). Vancouver: Blackwell Publishing.

[30] Ruscitti, F., et al. (2020). Quantification of lung fibrosis in IPF-like mouse model and pharmacological response to treatment by micro-computed tomography. Frontiers in Pharmacology, 11, article number 1117. doi: 10.3389/fphar.2020.01117.

[31] Savin, I.A., Zenkova, M.A., & Sen’kova, A.V. (2022). Pulmonary fibrosis as a result of acute lung inflammation: Molecular mechanisms, relevant in vivo models, prognostic and therapeutic approaches. International Journal of Molecular Sciences, 23(23), article number 14959. doi: 10.3390/ijms232314959.

[32] Scharm, S.C., et al. (2022). Increased regional ventilation as early imaging marker for future disease progression of interstitial lung disease: A feasibility study. European Radiology, 32(9), 6046-6057. doi: 10.1007/s00330-022-08702-w.

[33] Soldati, G., Smargiassi, A., Demi, L., & Inchingolo, R. (2020). Artifactual lung ultrasonography: It is a matter of traps, order, and disorder. Applied Sciences, 10(5), article number 1570. doi: 10.3390/app10051570.

[34] Stanel, S.C., & Rivera-Ortega, P. (2023). Present and future perspectives in early diagnosis and monitoring for progressive fibrosing interstitial lung diseases. Frontiers in Medicine, 10, article number 1114722. doi: 10.3389/fmed.2023.1114722.

[35] Tanabe, N., McDonough, J.E., Vasilescu, D.M., Ikezoe, K., Verleden, S.E., Xu, F., Wuyts, W.A., Vanaudenaerde, B.M., Colby, T.V., & Hogg, J.C. (2020). Pathology of idiopathic pulmonary fibrosis assessed by a combination of micro-computed tomography, histology, and immunohistochemistry. The American Journal of Pathology, 190(12), 2427-2435. doi: 10.1016/j.ajpath.2020.09.001.

[36] Thierry, F., Handel, I.G., Hammond, G., King, L.G., Corcoran, B., & Schwarz, T. (2017). Further characterization of computed tomographic and clinical features for staging and prognosis of idiopathic pulmonary fibrosis in West Highland white terriers. Veterinary Radiology & Ultrasound, 58(4), 381-388. doi: 10.1111/vru.12491.

[37] Vats, A., & Chaturvedi, P. (2023). The regenerative power of stem cells: Treating bleomycininduced lung fibrosis. Stem Cells and Cloning: Advances and Applications, 16, 43-59. doi: 10.2147/SCCAA.S419474.

[38] Wang, X.Z. (1992). The role of oxygen radicals in bleomycin-induced pulmonary fibrosisChinese Journal of Tuberculosis and Respiratory Diseases, 15(3), 158-191.

[39] Xiong, Y., et al. (2021). Dehydrocostus lactone inhibits BLM-induced pulmonary fibrosis and inflammation in mice via the JNK and p38 MAPK-mediated NF-κB signaling pathways. International Immunopharmacology, 98, article number 107780. doi: 10.1016/j.intimp.2021.107780.

[40] Zakaria, D.M., Zahran, N.M., Arafa, S.A., Mehanna, R.A., & Abdel-Moneim, R.A. (2020). Histological and physiological studies of the effect of bone marrow-derived mesenchymal stem cells on bleomycin induced lung fibrosis in adult albino rats. Tissue Engineering and Regenerative Medicine, 18(1), 127-141. doi: 10.1007/s13770-020-00294-0.

[41] Zhang, Y., Tu, B., Song, K., Lin, L., Liu, Z., Lu, D., Chen, Q., & Tao, H. (2023). Epigenetic hallmarks in pulmonary fibrosis: New advances and perspectives. Cellular Signalling, 110, article number 110842. doi: 10.1016/j.cellsig.2023.110842.