Comparative assessment of the hepatoprotective activity of genistein and silymarin in experimental carbon tetrachloride liver damage

Introduction. The search for new effective and safe hepatoprotectors is an urgent task in experimental toxicology and pharmacology. Natural polyphenols, particularly the isoflavone genistein, are considered promising compounds due to their antioxidant and anti-apoptotic properties.

The aim of this study is to experimentally evaluate the hepatoprotective efficacy of genistein in a model of acute toxic hepatitis induced by carbon tetrachloride (CCl₄).

Material and methods. The study was performed on 84 male white non-pedigree rats. Acute toxic hepatitis was simulated by a single intragastric administration of carbon tetrachloride (CCl₄) at a dose of 1 ml/kg. The hepatoprotective effect of genistein was studied by a single intragastric administration at a dose of 150 mg/kg 2 hours before CCl₄. Silymarin was used as a reference drug, which was also administered intragastrically at a dose of 150 mg/kg 2 hours before CCl₄. The animals were divided into four groups: biological control, CCl₄ administration, silymarin + CCl₄ administration, and genistein + CCl₄ administration. The effectiveness was assessed based on survival rates, body weight dynamics, liver weight, the propofol test, and the activity of ALT, AST, and alkaline phosphatase (ALP) in the blood serum. Statistical processing of the obtained data was performed using the GraphPad Prism 10 software. The statistical significance of differences in survival rates was assessed using the χ² test, and the Mann-Whitney U-test was used to assess quantitative indicators.

Results. CCl₄ administration led to the death of 13% of the animals, while silymarin reduced the mortality rate to 9%, while genistein provided 100% survival rate. Genistein statistically significantly reduced the activity of ALT on the 1^st and 3^rd days compared to the CCl₄ group and demonstrated better efficacy compared to silymarin in correcting cholestasis (reduction of ALP) and restoring the liver’s detoxification function (propofol test).

Limitations. The limitations of the study are due to the fact that the results were obtained in experiments on rats, which does not allow for direct extrapolation to humans. The work did not carry out a morphological assessment of the liver, any parameters of oxidative stress, inflammation, serum bilirubin and albumin, GGT activity or other additional markers of liver damage were not determined, which limits the completeness of the characteristics of the hepatoprotective effect of genistein.

Conclusion. Genistein has a hepatoprotective effect, exhibits a more pronounced effect compared to silymarin in a number of studied parameters and can be considered as a promising compound for further study in hepatology.

Compliance with ethical standards. The experimental study was approved by the local bioethics committee of the Center for Experimental Pharmacology at SPbSU (protocol No. Rats–01–FD-25 dated 06.10.25 and No. Rats–01–FD-26 dated 11.11.25).

Contribution of the authors:
Erlin G.V. – conducting the experiment; statistical processing and analysis of the results, writing text;
Karavaeva A.V. – biochemical analysis of rat serum samples;
Strelova O.Yu. – material analysis, editing;
Grebenyuk A.N. – the concept and design of the study; material analysis, editing.
All co-authors – approval of the final version of the article, responsibility for the integrity of all parts of the article.

Conflict of interests. The authors declare the absence of obvious and potential conflicts of interest in connection with the publication of this article.

Funding. The study had no sponsorship.

Received: April 23, 2026 / Revised: April 25, 2026 / Accepted: June 1, 2026 / Published: June 30, 2026

1. Niu C., Zhang J., Okolo P.I. The possible pathogenesis of liver fibrosis: therapeutic potential of natural polyphenols. Pharmacol. Rep. 2024; 76(5): 944–61. https://doi.org/10.1007/s43440-024-00638-w https://elibrary.ru/jockrs

2. Haleeva N.B., Arbuzov A.L. Romanov A.A. The experience of using the innovative medicinal hepatoprotector Gepafor with a probiotic component in the treatment of alcoholic liver damage. Vestnik medicinskogo instituta nepreryvnogo obrazovaniya. 2025; 2(5): 122-126 (in Russian)

3. Ayala-Calvillo E., Rodríguez-Fragoso L., Álvarez-Ayala E., Leija-Salas А. EGF-receptor phosphorylation and downstream signaling are activated by genistein during subacute liver damage. J. Mol. Histol. 2023; 54(3): 231–44. https://doi.org/10.1007/s10735-023-10127-8 https://elibrary.ru/agqekv

4. Erlin G.V., Strelova O.Yu., Grebenyuk A.N. Prospects for the use of genistein for the treatment of poisoning with hepatotoxic chemicals (literature review). Toksikologicheskii vestnik. 2024; 32(2): 63–71. https://elibrary.ru/rknhsm (in Russian)

5. Guidelines for Conducting Preclinical Drug Trials. Part One [Rukovodstvo po provedeniyu doklinicheskikh issledovanii lekarstvennykh sredstv: Chast’ pervaya]. Moscow: Grif i K; 2012. (in Russian)

6. Sukhanov D.S., Okovityi S.V., Yablonskyi P.K., Vinogradova T.I., Pavlova M.V. Hepatotropic therapy in treatment of liver injury. Antibiotiki i khimioterapiya. 2012; 57(5-6): 41–52. https://elibrary.ru/sclfud (in Russian)

7. Okovityi S.V., Raikhelson K.L., Prikhodko V.A. Combined hepatoprotective pharmacotherapy for liver disease. Eksperimental‘naya i klinicheskaya gastroenterologiya. 2022; (7): 5–20. https://doi.org/10.31146/1682-8658-ecg-203-7-5-20 https://elibrary.ru/ykqbbi (in Russian)

8. Vovk E.I. Drug-induced liver damage: silymarin. RMZh. 2017; 25(10): 750–3. https://elibrary.ru/zmrmtj (in Russian)

9. Al-Rasheed N., Faddah L., Al-Rasheed N., Bassiouni Y.A., Hasan I.H., Mahmoud A.M., et al. Protective effects of silymarin, alone or in combination with chlorogenic acid and/or melatonin, against carbon tetrachloride-induced hepatotoxicity. Pharmacogn. Mag. 2016; 12(Suppl. 3): S337–45. https://doi.org/10.4103/0973-1296.185765 https://elibrary.ru/yegsup

10. Chernova A.P., Shormanov V.K., Davydkina A.E. Propofol: use, toxicology and assay features. Sudebno-meditsinskaya ekspertiza. 2022; 65(5): 46–51. https://doi.org/10.17116/sudmed20226505146 https://elibrary.ru/gijmvl (in Russian)

11. Kravchenko L.V., Trusov N.V., Uskova M.A., Aksyonov I.V., Avrenyeva L.I., Guseva G.V., et al. Characterization of carbon tetrachloride acute toxicity as a model of oxidative stress. Toksikologicheskii vestnik. 2009; (1): 12–7. https://elibrary.ru/tpeyox (in Russian)

12. Golovko A.B., Yakovleva E.G. Dynamics of live weight of rats on the background of toxic experimental hepatitis. Vestnik Kurskoi gosudarstvennoi sel’skokhozyaistvennoi akademii. 2023; (9): 88–92. https://elibrary.ru/ydzkez (in Russian)

13. Chrungoo V.J., Singh K., Singh J. Silymarin mediated differential modulation of toxicity induced by carbon tetrachloride, paracetamol and D-galactosamine in freshly isolated rat hepatocytes. Indian J. Exp. Biol. 1997; 35(6): 611–7.

14. Nikitin N.S., Kuznecov S.L., Tyshko N.V. Microstructure of the liver in rats after administration of CCl4 against the background of vitamin and mineral deficiency. Morfologia.2019; 155(3): 42-7. (in Russian)

15. Satyam S.M., Bairy L.K., Rehman A., Attia M., Ahmed L., Emad K., et al. Unlocking synergistic hepatoprotection: dapagliflozin and silymarin combination therapy modulates nuclear erythroid 2-related factor 2/heme oxygenase-1 pathway in carbon tetrachloride-induced hepatotoxicity in Wistar rats. Biology (Basel). 2024; 13(7): 473. https://doi.org/10.3390/biology13070473 https://elibrary.ru/ahlsjp

16. Farruggio S., Cocomazzi G., Marotta P., Romito R., Surico D., Calamita G., et al. Genistein and 17β-Estradiol Protect Hepatocytes from Fatty Degeneration by Mechanisms Involving Mitochondria, Inflammasome and Kinases Activation. Cell. Physiol. Biochem. 2020; 54(3): 401–16. https://doi.org/10.33594/000000227 https://elibrary.ru/cvkxyw

17. Kamm D.R., McCommis K.S. Hepatic stellate cells in physiology and pathology. J. Physiol. 2022; 600(8): 1825–37. https://doi.org/10.1113/JP281061 https://elibrary.ru/vfwdtn

18. Campana L., Esser H., Huch M., Forbes S. Liver regeneration and inflammation: from fundamental science to clinical applications. Nat. Rev. Mol. Cell Biol. 2021; 22(9): 608–24. https://doi.org/10.1038/s41580-021-00373-7 https://elibrary.ru/ajpuvl

19. Borah L., Sen S., Mishra M., Barbhuiya P.A., Pathak M.P. Therapeutic potential of genistein: insights into multifaceted mechanisms and perspectives for human wellness. Curr. Top. Med. Chem. 2025; 25(27): 3190–202. https://doi.org/10.2174/0115680266377646250527075042 https://elibrary.ru/jidzhq

20. Tuli H.S., Tuorkey M.J., Thakral F., Sak K., Kumar M., Sharma A.K., et al. Molecular mechanisms of action of genistein in cancer: recent advances. Front. Pharmacol. 2019; 10: 1336. https://doi.org/10.3389/fphar.2019.01336

21. McGill M.R., Jaeschke H. Biomarkers of drug-induced liver injury: progress and utility in research, medicine, and regulation. Expert Rev. Mol. Diagn. 2018; 18(9): 797–807. https://doi.org/10.1080/14737159.2018.1508998 https://elibrary.ru/yjqxvz