Ukr.Biochem.J. 2021; Volume 93, Issue 5, Sep-Oct, pp. 72-81


Semicarbazide diminishes the signs of bleomycin-induced pulmonary fibrosis in rats

O. O. Hudkova*, I. P. Krysiuk, T. O. Kishko,
N. M. Popova, L. B. Drobot, N. V. Latyshko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;

Received: 16 July 2021; Accepted: 22 September  2021

The pathogenesis of pulmonary fibrosis (PF) is accompanied by extracellular matrix (ECM) deposition, oxidative stress, and inflammation progression, as well as hyperactivation of amine oxidases (AOs), which contribute to disease manifestation. The present study aims to elucidate the effect of semicarbazide (SC), an inhibitor of Cu-containing AOs: lysyl oxidase (LOX), semicarbazide sensitive amine oxidase (SSAO), diamine oxidase (DAO), on PF induced in rats by bleomycin (BLM). Eighteen male Wistar rats were randomly divided into four groups: Control, rats of BLM group received BLM (5 mg/kg, intratracheally once), BLM+SC group obtained 0.005% solution of SC (about 50 µg per capita per day) for three weeks starting immediately after BLM injection, and the Control+SC group drank the same solution as BLM+SC group. The content of cross-linked collagen in total bronchi and free radicals in lung, activities of LOX, SSAO, DAO, polyamine oxidase (PAO), Cu, Zn-superoxide dismutase (SOD1), catalase (CAT) and glutathione peroxidase (GPx) in lung and blood were measured. BLM injection induced PF that was confirmed histologically and morphometrically as well as by the elevation of the content of cross-linked collagen and free radicals. The activities of LOX and SSAO involved in post-translational modification of ECM and inflammation were significantly increased (P < 0.05). The activities of DAO, and PAO that control polyamine metabolism were also essentially raised. Among antioxidant enzymes, only GPx was activated in the BLM group as compared to control. These changes were absent in the BLM+SC group. SC intake promoted the fact that the histology and morphometric parameters of lung tissue, the content of cross-linked collagen in the bronchi and free radicals in the lung, as well as the activity of the studied enzymes remained at the control level. Our data suggest that SC suppresses the development of BLM-induced PF by inhibiting AOs activities.

Keywords: , , , , ,


  1. Clarke DL, Carruthers AM, Mustelin T, Murray LA. Matrix regulation of idiopathic pulmonary fibrosis: the role of enzymes. Fibrogenesis Tissue Repair. 2013;6(1):20. PubMed, PubMedCentral, CrossRef
  2. Cheresh P, Kim SJ, Tulasiram S, Kamp DW. Oxidative stress and pulmonary fibrosis. Biochim Biophys Acta. 2013;1832(7):1028-1040. PubMed, PubMedCentral, CrossRef
  3. Marttila-Ichihara F, Elima K, Auvinen K, Veres TZ, Rantakari P, Weston C, Miyasaka M, Adams D, Jalkanen S, Salmi M. Amine oxidase activity regulates the development of pulmonary fibrosis. FASEB J. 2017;31(6):2477-2491. PubMedCrossRef
  4. Structure and functions of amine oxidases. (Ed by Mondovi B). CRC Press. 2017. 299 p. CrossRef
  5. Wang J, Zhu Y, Tan J, Meng X, Xie H, Wang R. Lysyl oxidase promotes epithelial-to-mesenchymal transition during paraquat-induced pulmonary fibrosis. Mol Biosyst. 2016;12(2):499-507. PubMed, CrossRef
  6. Salmi M, Jalkanen S. Vascular Adhesion Protein-1: A Cell Surface Amine Oxidase in Translation. Antioxid Redox Signal. 2019;30(3):314-332. PubMed, PubMedCentral, CrossRef
  7. Lucarini L, Pini A, Rosa AC, Lanzi C, Durante M, Chazot PL, Krief S, Schreeb A, Stark H, Masini E. Role of histamine H4 receptor ligands in bleomycin-induced pulmonary fibrosis. Pharmacol Res. 2016;111:740-748. PubMed, CrossRef
  8. Branco ACCC, Yoshikawa FSY, Pietrobon AJ, Sato MN. Role of Histamine in Modulating the Immune Response and Inflammation. Mediators Inflamm. 2018;2018:9524075.  PubMed, PubMedCentral, CrossRef
  9. Hoet PH, Nemery B. Polyamines in the lung: polyamine uptake and polyamine-linked pathological or toxicological conditions. Am J Physiol Lung Cell Mol Physiol. 2000;278(3):L417-L433. PubMed, CrossRef
  10. Grasemann H, Shehnaz D, Enomoto M, Leadley M, Belik J, Ratjen F. L-ornithine derived polyamines in cystic fibrosis airways. PLoS One. 2012;7(10):e46618. PubMed, PubMedCentral, CrossRef
  11. Cox TR, Bird D, Bake AM, Barker HE, Ho MWY, Lang G, Erler JT. LOX-mediated collagen crosslinking is responsible for fibrosis-enhanced metastasis. Cancer Res. 2013;73(6):1721-1732. PubMed, PubMedCentral, CrossRef
  12. Reinert T, Baldotto CSR, Nunes FAP, Scheliga AAS. Bleomycin-Induced Lung Injury. J Cancer Res. 2013;2013:1-9.  CrossRef
  13. Latyshko N, Gudkova O, Dmytrenko M. Semicarbazide as potential source of formaldehyde and nitric oxide formation. Drugs Ther Stud. 2012;2(1):e9. CrossRef
  14. Gudkova OO, Latyshko NV, Gudkova LV, Mikhailovsky VO. Rat liver catalase under artificial hypobiosis conditions. Biopolym Cell. 2005;21(1):28-34. (In Ukrainian). CrossRef
  15. Korzhevsky DE, Gilyarov AV. Fundamentals of histological technique. St. Petersburg: SpetsLit. 2010. 96 p. (In Russian).
  16. Zaides AL, Mikhailov AN, Pushchenko OI. Modified method for determination of oxyproline. Biochemistry. 1964; 29(1): 5-7.
  17. Volodina TT, Dzvonkevych ND, Petrun’ LM, Krysiuk IP, Popova NM, Shandrenko SH, Dmytrenko MP. Altered collagene characteristics and lysyl oxidase activity in lathyrism. Fiziol Zh. 2011;57(3):62-68. (In Ukrainian). PubMed
  18. Gudkova OO, Latyshko NV, Shandrenko SG. Amine oxidases as important agents of pathological processes of rhabdomyolysis in rats. Ukr Biochem J. 2016;88(1):79-87. PubMed, CrossRef
  19. Gudkova OO, Latyshko NV, Zaitseva OV, Shandrenko SG. Purification procedure and assay for the activity of lysyl oxidase. Ukr Biochem J. 2018;90(5):98-105. CrossRef
  20. Labudzynskyi DO, Zaitseva OV, Latyshko NV, Gudkova OO, Veliky MM. Vitamin D(3) contribution to the regulation of oxidative metabolism in the liver of diabetic mice. Ukr Biochem J. 2015; 87(3):75-90. PubMed, CrossRef
  21. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72(1-2):248-254. PubMed, CrossRef
  22. Zhang CY, Duan JX, Yang HH , Sun CC, Zhong WJ, Tao JH, Guan XX, Jiang HL, Hammock BD, Hwang SH, Zhou Y, Guan CX. COX-2/sEH dual inhibitor PTUPB alleviates bleomycin-induced pulmonary fibrosis in mice via inhibiting senescence. FEBS J. 2020;287(8):1666-1680. PubMed, PubMedCentral, CrossRef
  23. Maranghi F, Tassinari R, Marcoccia D, Altieri I, Catone T, De Angelis G, Testai E, Mastrangelo S, Evandri MG, Bolle P, Lorenzetti S. The food contaminant semicarbazide acts as an endocrine disrupter: Evidence from an integrated in vivo/in vitro approach. Chem Biol Interact. 2010;183(1):40-48. PubMed, CrossRef
  24. Dmytrenko MP, Shandrenko SH, Petrun LM, Kishko TO, Sylonova NV, Latyshko NV, Gudkova OO, Sushkova VV. Formaldehyde metabolism in semicarbazide intoxication. Ukr Biokhim Zhurn. 2010;82(4):86-91. (In Ukrainian). PubMed
  25. Bailey AJ, Peach CM, Fowler LJ. Chemistry of the collagen cross-links. Isolation and characterization of two intermediate intermolecular cross-links in collagen. Biochem J. 1970;117(5):819-831. PubMed, PubMedCentral, CrossRef
  26. Mercier N. The role of ‘semicarbazide-sensitive amine oxidase’ in the arterial wall. Artery Res. 2009;3(4):141-147. CrossRef
  27. Giannoni E, Parri M, Chiarugi P. EMT and oxidative stress: a bidirectional interplay affecting tumor malignancy. Antioxid Redox Signal. 2012;16(11):1248-1263. PubMed, CrossRef
  28. Richter K, Kietzmann T. Reactive oxygen species and fibrosis: further evidence of a significant liaison. Cell Tissue Res. 2016;365(3):591-605. PubMed, PubMedCentral, CrossRef
  29. Oury TD, Thakker K, Menache M, Chang LY, Crapo JD, Day BJ. Attenuation of bleomycin-induced pulmonary fibrosis by a catalytic antioxidant metalloporphyrin. Am J Respir Cell Mol Biol. 2001;25(2):164-169. PubMed, CrossRef

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License.