Effects of triterpenoids in fatty products on liver condition of laboratory animals with acute toxic hepatitis
AbstractOne of the principles of prevention and non-medicamentous treatment of liver diseases, including hepatitis of different etiology, is the normalization of the diet through the consumption of food with physiologically active ingredients, in particular betulin, which helps to eliminate the causes of metabolic and oxidative disorders within liver cells.
The aim of the research was to assess in vivo the influence of triterpene alcohol betulin extracted from Betula pendula Roth. birch bark in fat-containing products (for example mayonnaise) on the blood biochemical parameters and liver morphological structure of rats with initiated acute toxic hepatitis.
Material and methods. Hepatoprotective and antioxidant activities of betulin as part of mayonnaise samples has been investigated in vivo on the model of toxic hepatitis initiated by carbon tetrachloride in male Wistar rats weighing 210–265 g. The animals were divided into 4 groups of 10 animals each: CG-1 – intact, CG-2 and MG – with carbon tetrachloride initiated toxic hepatitis. rats of the main groups were orally administered mayonnaise once a day at a dosage of 1 ml for 21 days after the formation of the model pathology: OG-1 with the added betulin (1 mg per 1 kg of body weight), OG-2 without betulin. Disorders of metabolic and oxidative processes in liver cells of animals were evaluated by biochemical indicators of blood plasma: the level of glucose, albumin, total cholesterol, triglycerides and urea and the activity of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and γ-glutamyltransferase. Oxidative stress in rats was estimated by the activity of catalase and superoxide dismutase in blood hemolysate (at a dilution of 1:200 and 1:10, respectively); the total prooxidant (in blood plasma) and total antioxidant (in blood hemolysate at a dilution of 1:10) activity were determined spectrophotometrically (colored complexes of TWIN-80 oxidation products with thiobarbituric acid). The morphological structure of rats’ liver was estimated by microscopy of prepared cuts of hepatic tissue.
Results. Based on biochemical parameters of rat blood plasma, it has been established that the administration of mayonnaise with betulin prevents the development of cytolic syndrome and suppresses the process of peroxidation by directly neutralizing free radicals. Aspartate aminotransferase and alkaline phosphatase activity in blood plasma of the experimental animals of the main group MG-1 reduced by 20.7 and 35.2% compared with indicators of the rats of the main group MG-2. Glucose concentration normalized to the level of the control group CG-1. The concentration of bilirubin and triglycerides decreased by 22.9 and by 48.1%, which indicates a significant reduction in the indicators of cholestatic syndrome in the group of animals OG-1 compared to OG-2. The total prooxidant activity and the concentration of thiobarbiturate-reactive products decreased compared to the CG-2 and MG-2 groups, which indicates the suppression of oxidative stress and, as a result, an improvement in liver conditions of animals with toxic hepatitis even when taking a fat-containing product. In liver histopeparates of animals receiving mayonnaise with betulin, necrobotic changes were less pronounced in comparison with the group MG-2. They were estimated at 1 point: small-drip dystrophy spots were found, haemorrhages in the interregional septum with inflammatory infiltration in the course of hemorrhages against the presence of necrosis hepatocytes with pronounced adipose dystrophy in the centres of the lobules, step necrosis with signs of replacing the damaged hepatocytes of the connective tissue, accompanied by centrolobular hemorrhages in MG-2 rats.
Conclusion. Introduced into the composition of mayonnaise betulin, reduces the development of cytolic syndrome in toxic hepatitis and suppresses the process of peroxidation, on the basis of which fat-containing foods with betulin can be recommended for clinical examination as specialized products in acute and chronic liver diseases, including complicated cholestasis.
Keywords:betulin; mayonnaise; toxic hepatitis; rats; blood chemistry; antioxidant activity; liver morphology
Funding. This work was supported by the project 075-00316-20-01 (FZMM-2020-0013, mnemocode 0611-2020-013) from the Ministry of Science and Higher Education of the Russian Federation.
Conflict of interest. The authors declare no conflict of interest.
Contribution. The concept and design of the study – Averyanova E.V., Shkolnikova M.N.; data collection and processing – Mazko O.N.; statistical data processing – Chugunova O.V.; writing the text – Shkolnikova M.N., Averyanova E.V.; editing, approval of the final version of the article, responsibility for the integrity of all parts of the article – all authors.
For citation: Averyanova E.V., Shkolnikova M.N., Chugunova O.V., Mazko O.N. Effects of triterpenoids in fatty products on liver condition of laboratory animals with acute toxic hepatitis. Voprosy pitaniia [Problems of Nutrition]. 2023; 92 (4): 81–91. DOI: https://doi.org/10.33029/0042-8833-2023-92-4-81-91 (in Russian)
References
1. Schenk A., Ghallab A., Hofmann U., Hassan R., Schwarz M., Schuppert A., et al. Physiologically-based modelling in mice suggests an aggravated loss of clearance capacity after toxic liver damage. Sci Rep. 2017; 7 (1): 6224. DOI: https://doi.org/10.1038/s41598-017-04574-z
2. Balukova E.V., Uspensky Yu.P., Fominykh Yu.A. Liver diseases of various genesis (toxic, drug-induced, dysmetabolic): from etiological heterogenicity to a single unified therapy of patients. RMZh. Meditsinskoe obozrenie [RMJ. Medical Review]. 2018; 2 (1-1): 35–40. (in Russian)
3. Gubergrits N.B., Belyaeva N.V., Klochkov A.E., Lukashevich G.M., Fomenko P.G. Medicinal lesions of the liver: from pathogenesis to treatment. Vestnik kluba pankreatologov [Bulletin of the Pancreatologists Club]. 2020; 11 (46): 72–80. DOI: https://doi.org/10.33149/vkp.2020.01.10 (in Russian)
4. Gubergrits N.B., Belyaeva N.V., Klochkov A.Y., Lukashevish G.M., Fomenko P.G. Modern views on nutrition and physical activity in the treatment of non-alcoholic fatty liver disease. Eksperimentalnaya i klinicheskaya gastoenterologiya [Experimental and Clinical Gastroenterology]. 2018; (2): 100–9. (in Russian)
5. Sasunova A.N., Morozov S.V., Sobolev R.V., Isakov V.A., Kochetkova A.A., Vorob’eva I.S. Efficacy of newly developed food for special dietary use in the diet of patients with non-alcoholic steatohepatitis. Voprosy pitaniia [Problems of Nutrition]. 2022; 91 (2): 31–42. DOI: https://doi.org/10.33029/0042-8833-2022-91-2-31-42 (in Russian)
6. Official website. All oils of the world. URL: https://www.oilworld.ru/analytics/localmarket/334695 (date of access March 05, 2023). (in Russian)
7. Labetsky V.V., Feofilaktova O.V. Research of rheological parameters of emulsion sauces. Pishchevaya promyshlennost’: nauka i tehnologii [Food Industry: Science and Technology]. 2022; 15 (3): 86–9. DOI: https://doi.org/10.47612/2073-4794-2022-15-3(57)-86-89 (in Russian)
8. Vorob’eva V.M., Vorob’eva I.S., Morozov S.V., Sasunova A.N., Kochetkova A.A., Isakov V.A. Specialized food products for dietary correction of the diet of patients with nonalcoholic steatohepatitis. Voprosy pitaniia [Problems of Nutrition]. 2021; 90 (2): 100–9. DOI: https://doi.org/10.33029/0042-8833-2021-90-2-100-109 (in Russian)
9. Morisco F., Vitaglione P., Amoruso D., Russo B., Fogliano V., Capo-raso N. Foods and liver health. Mol Aspects Med. 2008; 29 (1–2): 144–50. DOI: https://doi.org/10.1016/j.mam.2007.09.003
10. Ma C., Wang C., Zhang Y., Zhou H., Li Y. Potential natural compounds for the prevention and treatment of nonalcoholic fatty liver disease: a review on molecular mechanisms. Curr Mol Pharmacol. 2022; 15 (6): 846–61. DOI: https://doi.org/10.2174/1874467215666211217120448
11. Cerletti C., Colucci M., Storto M., Semeraro F., Ammollo C.T., Incampo F., et al. Randomised trial of chronic supplementation with a nutraceutical mixture in subjects with non-alcoholic fatty liver disease. Br J Nutr. 2020; 123 (2): 190–7. DOI: https://doi.org/10.1017/S0007114519002484
12. Inozemtsev P.O., Fedorova L.I., Lepekhova S.A. Modern methods of correction and prevention of liver failure. Effektivnaya farmakoterapiya [Effective Pharmacotherapy]. 2020; 16 (1): 46–51. DOI: https://doi.org/10.33978/2307-3586-2020-16-1-46-51 (in Russian)
13. Chan C.-C., Lee K.-C., Huang Y.-H., Chou C.K., Lin H.C., Lee F.Y. Regulation by resveratrol of the cellular factors mediating liver damage and regeneration after acute toxic liver injury. J Gastroenterol Hepatol. 2014; 29 (3): 603–13. DOI: https://doi.org/10.1111/jgh.12366
14. García-Niño W. R., Pedraza-Chaverrí J. Protective effect of curcumin against heavy metals-induced liver damage. Food Chem Toxicol. 2014; 69: 182–201. DOI: https://doi.org/10.1016/j.fct.2014.04.016
15. Mirzanajafi-Zanjani M., Yousefi M., Ehsani A. Challenges and approaches for production of a healthy and functional mayonnaise sauce. Food Sci Nutr. 2019; 7 (8): 2471–84. DOI: https://doi.org/10.1002/fsn3.1132
16. Fenoglio D., Soto M-D., Alarcon M-J., Ferrario M., Guerrero S., Matiacevich S. Active food additive based on encapsulated yerba mate (Ilex paraguariensis) extract: effect of drying methods on the oxidative stability of a real food matrix (mayonnaise). J Food Sci Technol. 2021; 58: 1574–84. DOI: https://doi.org/10.1007/s13197-020-04669-y
17. Zhang W., Jiang H., Yang J., Jin M., Du Y., Sun Q., Xu H. Safety assessment and antioxidant evaluation of betulin by LC-MS combined with free radical assays. Anal Biochem. 2019; 587: 113460. DOI: https://doi.org/10.1016/j.ab.2019.113460
18. Amiri S., Dastghaib S., Ahmadi M., Mehrbod P., Khadem F., Behrouj H., et al. Betulin and its derivatives as novel compounds with different pharmacological effects. Biotechnol Adv. 2020; 38: 107409. DOI: https://doi.org/10.1016/j.biotechadv.2019.06.008
19. Boparai A., Niazi J., Bajwa N., Singh P.A. Betulin a pentacyclic triterpenoid: an hour to rethink the compound. Open Access J Transl Med Res. 2017; 1 (2): 53–9. DOI: https://doi.org/10.15406/oajtmr.2017.01.00012
20. Vorob’eva O.A., Malygina D.S., Grubova E.V., Mel’nikova N.B. Betulin derivatives. Biological activity and solubility enhancement. Khimiya rastitel’nogo syr’ya [Chemistry of Plant Raw Material]. 2019; (4): 407–30. DOI: https://doi.org/10.14258/jcprm.2019045419 (in Russian)
21. Hordyjewska A., Ostapiuk A., Horecka A., Kurzepa J. Betulin and betulinic acid: Triterpenoids derivatives with a powerful biological potential. Phytochem Rev. 2019; 18: 929–51. DOI: https://doi.org/10.1007/s11101-019-09623-1
22. Nootropic: pat. 2300389/C1 Russian Federation. No. 2005138574/15. Kovalev G.I., Salimov R.M., Balakshin V.V., Chistyakov A.N.; application. 12.12.2005; publ. 10.06.2007, Bull. No. 16: 17 p. (in Russian)
23. A remedy for the prevention and treatment of Parkinson’s disease: pat. 2324492/C1 Russian Federation. No. 2006142767/15. Kovalev G.I., Abaimov D.A., Firstova Yu.Yu., Balakshin V.V., Chistyakov A.N.; application. 04.12.2006; publ. 20.05.2008, Bull. No. 14: 27 p. (in Russian)
24. Rastogi S., Pandey M.M., Kumar Singh Rawat A. Medicinal plants of the genus Betula – traditional uses and a phytochemical-pharmacological review. J Ethnopharmacol. 2015; 159: 62–83. DOI: https://doi.org/10.1016/j.jep.2014.11.010
25. Chen H., Xiao H., Pang J. Parameter optimization and potential bioactivity evaluation of a betulin extract from white birch bark. Plants. 2020; 9 (3): 392–407. DOI: https://doi.org/10.3390/plants9030392
26. Shapekova N.L., Aymakov O.A., Safarov R.Z., Salmanov G.A. Bioactivity of betulin, betulin aldehyde and their derivatives. In: Materials of the scientific and practical conference «Global Science and Innovations IV». Sofia, 2018: 122–30. (in Russian)
27. Furtado J.C., Pirson N., Edelberg L., Miranda H.M., Loira-Pastoriza L., Preat C. Pentacyclic triterpene bioavailability: an overview of in vitro and in vivo studies. Molecules. 2017; 22 (3): 400. DOI: https://doi.org/10.3390/molecules22030400
28. Method of obtaining betulin: pat. 2640587C1 Russian Federation. No. 2017104346. Averyanova E.V., Shkolnikova M.N., Tsyganok S.N., Khmelev V.N., Shakura V.A.; application. 09.02.2017; publ. 10.01.2018, Bull. No. 1: 10 p. (in Russian)
29. Kuznetsova S.A., Skvortsova G.P., Malyar Yu.N., Skurydina E.S., Veselova O.F. Extraction betulin from birch bark and study of its physicochemical and pharmacological properties. Khimiya rastitel’nogo syr’ya [Chemistry of Plant Raw Material]. 2013; (2): 93–100. DOI: https://doi.org/10.14258/jcprm.201302093 (in Russian)
30. Aver’yanova E.V., Shkol’nikova M.N., Chugunova O.V. Antioxidant properties of triterpenoids in fat-containing products. Tekhnika i tekhnologiya pischevykh proizvodstv [Technique and Technology of Food Production]. 2022; 52 (2): 233–43. DOI: https://doi.org/10.21603/2074-9414-2022-2-2358 (in Russian)
31. Skuratov A.G. Tetrachloromethane model of hepatitis and cirrhosis in rats. Eksperimental’naya i klinicheskaya gastoenterologiya [Experimental and Clinical Gastroenterology]. 2012; (9): 37–40. (in Russian)
32. Guidelines for experimental (preclinical) study of new pharmacological substances. Edited by Khabriev R.U. 2nd ed. Moscow: Meditsina, 2005: 832 p. ISBN: 5-225-04219-8. (in Russian)
33. Makarova M.A., Baranova I.A. Main hepatic syndromes in practice of internist. Consilium Medicum. 2017; 19 (8): 69–74. DOI: https://doi.org/10.26442/2075-1753_19.8.69-74 (in Russian)
34. Bryukhanov V.M., Zverev Ya.F., Lampatov V.V., Zharikov A.Yu., Talalaeva O.S. Methods of pre-clinical (experimental) study of the influence of drugs on renal function. Novosibirsk: GEO: 82 p. ISBN 978-5-906284-12-9. (in Russian)
35. Frolov B.A., Kalinina O.V., Kirillova A.V., Shtil’ A.A. Overcoming methotrexate induced liver toxicity: a role of triterpenoids. Klinicheskaya onkogematologiya. Fundamental’nye issledovaniya i klinicheskaya praktika [Clinical Oncohematology. Basic Research and Clinical Practice]. 2013; 6 (1): 1–10. (in Russian)
36. Vorob’eva O.A., Malygina D.S., Solov’eva A.G., Belyaeva K.L., Grubova E.V., Mel’nikova N.B. Antioxidant and pro-oxidant properties of betulin derivatives. Bioradikaly i antioksidanty [Bioradicals and Antioxidants]. 2018; 5 (4): 9–20. (in Russian)
37. Voronova O.A., Plotnikov E.V., Kalieva S.S., Nurpeyis E.E., Mamaeva E.A., Tashenov A.K., et al. Investigation of antioxidant activity representatives of triterpenoids series of lupane and oleane by using voltammetry. Vestnik Karagandinskogo universiteta. Seriya: Khimiya [Bulletin of Karaganda University. Series Chemistry]. 2017; (3): 31–7. (in Russian)
38. Yakovleva M.P., Medvedeva N.I., Saitov K.M., Ishmuratov G.Yu. Chemical behavior of betulin derivatives in reactions with organic nadacids. Vestnik Bashkirskogo universiteta [Bulletin of Bashkir University]. 2022; 27 (2): 323–9. DOI: https://doi.org/10.33184/bulletin-bsu-2022.2.12 (in Russian)
39. Tsymbal I.N. Regularities of the antioxidant effect of natural and synthetic triterpenoids of the lupane and β-amyrin series. Tyumen’: TyumGNGU, 2004: 24 p. (in Russian)