Scientific Reports of the National University of Life and Environmental Sciences of Ukraine

Corrective efficacy of milk phospholipids in tetracycline-induced changes in the amino acid composition of blood in rats

Serhii Bryzhenko, Viktoriia Gryshchenko
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Abstract

The aim of the study was to establish the patterns of changes in the amino acid spectrum of whole blood in laboratory rats against the background of tetracycline-induced hepatopathy and the administration of milk phospholipids. To experimentally reproduce a model of acute hepatopathy, laboratory rats were intragastrically administered tetracycline hydrochloride at a dose of 250 mg/kg body weight for 7 days. Four groups were formed, including rats that were: clinically healthy; affected by tetracycline-induced hepatosis (without treatment); affected by tetracycline-induced hepatosis and administered milk phospholipids; clinically healthy and administered milk phospholipids. The amino acid profile of whole blood was determined using paper chromatography with ninhydrin detection. The development of an imbalance in the amino acid profile of whole blood in rats as a result of the cytotoxic effect of tetracycline hydrochloride on hepatocytes was experimentally established. In animals with experimental hepatopathy under conditions of spontaneous recovery, a significant increase in the level of the heterogeneous histidine/taurine/asparagine fraction (by 54%) was observed in whole blood, along with a tendency towards increased levels of the combined cystine/cysteine, glycine/serine, and ornithine cycle amino acid fractions. The concentration of individual amino acids remained relatively stable. Administration of the phospholipid dietary supplement to diseased animals was accompanied by a pronounced mobilisation of amino acids in the blood within the combined fractions of cysteine/cystine (by 161%), arginine/ornithine/lysine (by 69%), histidine/taurine/asparagine (by 196%), glycine/serine/glutamine/aspartic acid (by 92%), and valine/norvaline/tryptophan (by 86%), indicating stimulation of nitrogen metabolism and ureagenesis and reflecting activation of detoxification processes, antioxidant defence, and the hepatotropic effect of phospholipid correction. The obtained results may be used for the diagnosis of metabolic disturbances related to intermediate amino acid metabolism in drug-induced hepatopathies and as markers for evaluating the efficacy of newly developed medicinal products

Keywords

liver; hepatopathy; fatty hepatosis model; corrective therapy; paper chromatography; marker changes; drug-induced complications

Suggested citation
Bryzhenko, S., & Gryshchenko, V. (2026). Corrective efficacy of milk phospholipids in tetracycline-induced changes in the amino acid composition of blood in rats. Scientific Reports of the National University of Life and Environmental Sciences of Ukraine, 22(3),117-135. https://doi.org/10.31548/dopovidi/3.2026.117
References
  1. Aghaei, F., Arabzadeh, E., Mahmoodzadeh Hosseini, H., & Shirvani, H. (2023). Exercise training and probiotic Lacticaseibacillus rhamnosus GG reduce tetracycline-induced liver oxidative stress and inflammation in rats with hepatic steatosis. Probiotics and Antimicrobial Proteins, 15(5), 1393-1405. doi: 10.1007/s12602-022-09994-6.
  2. Allison, R., Guraka, A., Shawa, I.T., Tripathi, G., Moritz, W., & Kermanizadeh, A. (2023). Drug induced liver injury – a 2023 update. Journal of Toxicology and Environmental Health. Part B, Critical Reviews, 26(8), 442-467. doi: 10.1080/10937404.2023.2261848.
  3. Amanatidou, A.I., et al. (2023). Plasma amino acids in NAFLD patients with obesity are associated with steatosis and fibrosis: Results from the MAST4HEALTH study. Metabolites, 13(8), article number 959. doi: 10.3390/metabo13080959.
  4. Baliou, S., Adamaki, M., Ioannou, P., Pappa, A., Panayiotidis, M.I., Spandidos, D.A., Christodoulou, I., Kyriakopoulos, A.M., & Zoumpourlis, V. (2021). Protective role of taurine against oxidative stress (Review). Molecular Medicine Reports, 24(2), article number 605. doi: 10.3892/mmr.2021.12242.
  5. Block, R.J., Durrum, E.L., & Zweig, G. (1955). A manual of paper chromatography and paper electrophoresis. New York: Academic Press.
  6. Borgne-Sanchez, A., & Fromenty, B. (2025). Mitochondrial dysfunction in drug-induced hepatic steatosis: Recent findings and current concept. Clinics and Research in Hepatology and Gastroenterology, 49(3), article number 102529. doi: 10.1016/j.clinre.2025.102529.
  7. Bryzhenko, S., & Gryshchenko, V. (2026). Amino acids of the biliary system in rats with tetracycline-induced hepatosis and the use of milk phospholipids. Ukrainian Journal of Veterinary Sciences, 17(1), 47-71. doi: 10.31548/veterinary1.2026.47.
  8. Cesarini, L., Grignaffini, F., Alisi, A., & Pastore, A. (2024). Alterations in glutathione redox homeostasis in metabolic dysfunction-associated fatty liver disease: A systematic review. Antioxidants, 13(12), article number 1461. doi: 10.3390/antiox13121461.
  9. Consden, R., Gordon, A.H., & Martin, A.J.P. (1944). Qualitative analysis of proteins: A partition chromatographic method using paper. Biochemical Journal, 38(3), 224-232. doi: 10.1042/bj0380224.
  10.  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/eli/dir/2010/63/oj/eng.
  11. Ghanem, S.E., et al. (2022). Evaluation of amino acids profile as non-invasive biomarkers of hepatocellular carcinoma in Egyptians. Tropical Medicine and Infectious Disease, 7(12), article number 437. doi: 10.3390/tropicalmed7120437.
  12. Gu, K., et al. (2023). Tryptophan alleviates lipopolysaccharide-induced liver injury and inflammation by modulating necroptosis and pyroptosis signaling pathways in piglets. Animal Biotechnology, 34(8), 4069-4080. doi: 10.1080/10495398.2023.2255064.
  13. Hackman, R.H., & Lazarus, M. (1956). Quantitative analysis of amino acids using paper chromatography. Australian Journal of Biological Sciences, 9(2), 281-292. doi: 10.1071/BI9560281.
  14. Han, S.K., Baik, S.K., & Kim, M.Y. (2025). Protein and macronutrient metabolism in liver cirrhosis: About sarcopenia. Nutrients, 17(21), article number 3346. doi: 10.3390/nu17213346.
  15. Kinny-Köster, B., Bartels, M., Becker, S., Scholz, M., Thiery, J., & Ceglarek, U. (2016). Plasma amino acid concentrations predict mortality in patients with end-stage liver disease. Plos One, 11(7), article number e0159205. doi: 10.1371/journal.pone.0159205.
  16. Korolova, D., Gryshchenko, V., Chernyshenko, T., Platonov, O., Hornytska, O., Chernyshenko, V., Klymenko, P., Reshetnik, Y., & Platonova, T. (2023). Blood coagulation factors and platelet response to drug-induced hepatitis and hepatosis in rats. Animal Models and Experimental Medicine, 6(1), 66-73. doi: 10.1002/ame2.12301.
  17.  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.
  18. Leonetti, S., Herzog, R.I., Caprio, S., Santoro, N., & Tricò, D. (2020). Glutamate-serine-glycine index: A novel potential biomarker in pediatric non-alcoholic fatty liver disease. Children, 7(12), article number 270. doi: 10.3390/children7120270.
  19. Li, Z., et al. (2020). Methionine metabolism in chronic liver diseases: An update on molecular mechanism and therapeutic implication. Signal Transduction and Targeted Therapy, 5, article number 280. doi: 10.1038/s41392-020-00349-7.
  20. Melnychuk, D., Hryshchenko, V., & Litvinenko, O. (2009). Veterinary bioactive additive of liposomal form and method of reparative therapy in hepatology. (Patent of Ukraine No. 86516). Retrieved from https://sis.nipo.gov.ua/uk/search/detail/422804/.
  21. Moore, S., & Stein, W.H. (1948). Photometric ninhydrin method for use in the chromatography of amino acids. Journal of Biological Chemistry, 176(1), 367-388. doi: 10.1016/S0021-9258(18)51034-6.
  22.  Order of the Ministry of Education and Science, Youth and Sports of Ukraine No. 249 “On Approval of the Procedure for Conducting Research and Experiments on Animals by Scientific Institutions”. (2012, March). Retrieved from https://zakon.rada.gov.ua/laws/show/z0416-12#Text.
  23. Partridge, S.M. (1948). Filter-paper partition chromatography of sugars: I. General description and application to the qualitative analysis of sugars in apple juice, egg white and foetal blood of sheep. Biochemical Journal, 42(2), 238-250. doi: 10.1042/bj0420238.
  24. Prechtl, L., et al. (2024). Circulating amino acid signature features urea cycle alterations associated with coronary artery disease. Scientific Reports, 14, article number 25848. doi: 10.1038/s41598-024-76835-7.
  25. Saitoh, W., Yamauchi, S., Watanabe, K., Takasaki, W., & Mori, K. (2014). Metabolomic analysis of arginine metabolism in acute hepatic injury in rats. Journal of Toxicological Sciences, 39(1), 41-50. doi: 10.2131/jts.39.41.
  26. Sun, Y., Demagny, H., Faure, A., Pontanari, F., Jalil, A., Bresciani, N., Yildiz, E., Korbelius, M., Perino, A., & Schoonjans, K. (2023). Asparagine protects pericentral hepatocytes during acute liver injury. Journal of Clinical Investigation, 133(7), article number e163508. doi: 10.1172/JCI163508.
  27. Tan, Q., Deng, S., & Xiong, L. (2025). Role of kynurenine and its derivatives in liver diseases: Recent advances and future clinical perspectives. International Journal of Molecular Sciences, 26(3), article number 968. doi: 10.3390/ijms26030968.
  28. Tang, Y., Chen, X., Chen, Q., Xiao, J., Mi, J., Liu, Q., You, Y., Chen, Y., & Ling, W. (2022). Association of serum methionine metabolites with non-alcoholic fatty liver disease: A cross-sectional study. Nutrition & Metabolism, 19, article number 21. doi: 10.1186/s12986-022-00647-7.
  29. Wang, Y., Li, J., Matye, D., Zhang, Y., Dennis, K., Ding, W.-X., & Li, T. (2018). Bile acids regulate cysteine catabolism and glutathione regeneration to modulate hepatic sensitivity to oxidative injury. JCI Insight, 3(8), article number e99676. doi: 10.1172/jci.insight.99676.
  30. Williams, R.J., & Kirby, H. (1948). Paper chromatography using capillary ascent. Science, 107(2784), 481-483. doi: 10.1126/science.107.2784.481.
  31. Yamakado, M., et al. (2017). Plasma amino acid profile associated with fatty liver disease and co-occurrence of metabolic risk factors. Scientific Reports, 7, article number 14485. doi: 10.1038/s41598-017-14974-w.
  32. Yan, Y., Chen, Q., Dai, X., Xiang, Z., Long, Z., Wu, Y., Jiang, H., Zou, J., Wang, M., & Zhu, Z. (2024). Amino acid metabolomics and machine learning for assessment of post-hepatectomy liver regeneration. Frontiers in Pharmacology, 15, article number 1345099. doi: 10.3389/fphar.2024.1345099.
  33. Yao, X., Cheng, Z., Agathokleous, E., Wei, Y., Feng, X., Li, H., Zhang, T., Li, S., Dhawan, G., & Luo, X.-S. (2024). Tetracycline and sulfadiazine toxicity in human liver cells Huh-7. Environmental Pollution, 345, article number 123454. doi: 10.1016/j.envpol.2024.123454.