Treatment of animals with fatty liver disease using a drug based on the seeds of Silybum marianum

  • V. Vlizlo Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies
  • O. Prystupa Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies
  • L. Slivinska Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies
  • B. Gutyj Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies
  • I. Maksymovych Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies
  • A. Shcherbatyy Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies
  • M. Lychuk Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies
  • U. Partyka Andrei Krupynskyi Lviv Medical Academy
  • B. Chernushkin Stepan Gzhytskyi National University of Veterinary
  • V. Rusyn Stepan Gzhytskyi National University of Veterinary
  • M. Leno Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies
  • K. Leskiv Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies
Keywords: rats; tetrachloromethane; Silybum marianum; liposomal drug; treatment

Abstract

Medicinal plants are a source of various therapeutic preparations. Therefore, the aim of our work was to prepare a liposomal drug from extract from the seeds of Silybum marianum (L.) Gaertn., adding tocopherol acetate, lecithin, squalene, and Tween 80. The drug was used on the laboratory animals (rats) intramuscularly to measure the efficacy of treatment of experimentally modeled toxic fatty liver disease. The fatty infiltration of the liver in the rats was caused by tetrachloromethane (ССl4). The efficacy of the liposomal drug based on the extract from S. marianum seeds was studied on 25 animals in which the liver pathology had been caused by 50% oil solution of ССl4, administered in the dose of 5 mL per kg of body weight. The diseased rats were divided into five groups, each consisting five animals. Animals of the four experimental groups – first, second, third, and fourth - received the drug intramuscularly in the doses of 0.05, 0.25, 0.50, and 1.50 mL/kg of body weight three times every two days, respectively. At the same time, the control rats received three-time intramuscular injection of physiological solution in the dose of 0.5 mL/kg of body weight. Treatment of the animals with fatty liver disease by injections of the drug based on the extract from S. marianum seeds normalized the general condition, significantly improved the functions and structure of the liver. Biochemical studies of blood serum of the sick animals after the treatment revealed increase in albumin content, which may suggest reduction of the protein-synthesizing function of the liver. The normalization of the bile-forming and bile-excreting functions of the liver, and also elimination of cholestasis were evidenced by reduced contents of bile acids and total bilirubin and increased total cholesterol in the blood serum of the rats. After treating the animals with the created drug, we saw decrease in the activity of the liver-indicator enzymes (aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, and glutamate dehydrogenase) in the blood serum, which is a sign of recovery of the structure of hepatocytes and elimination of cytolysis. Histological studies of the liver of the treated rats confirmed the positive effect of the liposomal drug on the organ’s structure. In the future studies, we plan to test this combination of agents in treatment of agricultural and domestic animals with liver pathologies.

References

Abenavoli, L., Izzo, A. A., Milić, N., Cicala, C., Santini A., & Capasso, R. (2018). Milk thistle (Silybum marianum): A concise overview on its chemistry, pharmacological, and nutraceutical uses in liver diseases. Phytotherapy Research, 32(11), 2202–2213.
Adetuyi, B. O., Omolabi, F. K., Olajide, P. A., & Oloke, J. K. (2021). Pharmacological, biochemical and therapeutic potential of milk thistle (Silymarin): A review. World News of Natural Sciences, 37, 75–91.
Armanini, E. H., Boiago, M. M., Cécere, B. G. O., Oliveira, P. V., Teixeira, C. J. S., Strapazzon, J. V., Bottari, N. B., Silva, A. D., Fracasso, M., Vendruscolo, R. G., Wagner, R., Gloria, E. M. D., Horn, V. W., Mendes, R. E., Baldissera, M. D., Vedovatto, M., & Da Silva, A. S. (2021). Protective effects of silymarin in broiler feed contaminated by mycotoxins: growth performance, meat antioxidant status, and fatty acid profiles. Tropical Animal Health and Production, 53(4), 442.
Bashchenko, M. I., Boiko, О. V., Honchar, О. F., Gutyj, B. V., Lesyk, Y. V., Ostapyuk, A. Y., Kovalchuk, І. І., & Leskiv, K. Y. (2020). The effect of milk thistle, metiphen, and silimevit on the protein-synthesizing function of the liver of lay ing hens in experimental chronic cadmium toxicosis. Ukrainian Journal of Ecology, 10(6), 164–168.
Bencze-Nagy, J., Strifler, P., Horváth, B., Such, N., Farkas, V., Dublecz, K., & Pál, L. (2023). Effects of dietary milk thistle (Silybum marianum) supplementation in ducks fed mycotoxin-contaminated diets. Veterinary Sciences, 10(2), 100.
Chernushkin, B. O., Vlizlo, V. V., Slivinska, L. G., Gutyj, B. V., Shcherbatyy, A. R., Maksymovych, I. A., Leno, M. I., Rusyn, V. I., Lychuk, M. H., Fedorovych, V. L., Lukashchuk, B. O., Zinko, H. O., & Prystupa, O. I. (2020). Treatment strategies for sheep with acute yellow athrophy of the liver caused by the fasciolosis. Ukrainian Journal of Ecology, 10(2), 294–301.
Curcio, A., Romano, A., Cuozzo, S., Nicola, A. D., Grassi, O., Schiaroli, D., Nocera, G. F., & Pironti, M. (2020). Silymarin in combination with vitamin C, vitamin E, coenzyme Q10 and selenomethionine to improve liver enzymes and blood lipid profile in NAFLD patients. Medicina (Kaunas), 56(10), 544.
Denev, S., Sotirov, L., Chobanova, S., Koynarski, T., Ivanov, V., Bozakova, N., & Stoev, S. (2020). Effect of silymarin and ochratoxin A on humoral natural immunity of broiler chickens. Journal of Central European Agriculture, 21(3), 492–498.
Dočkalová, H., Horký, P., Zeman, L., & Skládanka, J. (2018). Influence of milk thistle pressed parts on rats liver histology. Potravinarstvo Slovak Journal of Food Sciences, 12(1), 33–39.
Egresi, A., Sule, K., Szentmihalyi, K., Blazovics, A., Feher, E., Hagymasi, K., & Febel, H. (2020). Impact of milk thistle (Silybum marianum) on the mycotoxin caused redox-homeostasis imbalance of ducks liver. Toxicon, 187, 181–187.
El-Sheshtawy, S. M., El-Zoghby, A. F., Shawky, N. A., & Samak, D. H. (2021). Aflatoxicosis in Pekin duckling and the effects of treatments with lycopene and silymarin. Veterinary World, 14(3), 788–793.
Federico, A., Dallio, M., Masarone, M., Gravina, A. G., Di Sarno, R., Tuccillo, C., Cossiga, V., Lama, S., Stiuso, P., Morisco, F., Persico, M., & Loguercio, C. (2019). Evaluation of the effect derived from silybin with vitamin D and vitamin E administration on clinical, metabolic, endothelial dysfunction, oxidative stress parameters, and serological worsening markers in nonalcoholic fatty liver disease patients. Oxidative Medicine and Cellular Longevity, 2019, 8742075.
Fitzgerald, M., Heinrich, M., & Booker, A. (2020). Medicinal plant analysis: A historical and regional discussion of emergent complex techniques. Frontiers in Pharmacology, 10, 1480.
Galli, F., Bonomini, M., Bartolini, D., Zatini, L., Reboldi, G., Marcantonini, G., Gentile, G., Sirolli, V., & Di Pietro, N. (2022). Vitamin E (alpha-tocopherol) metabolism and nutrition in chronic kidney disease. Antioxidants, 11(5), 989.
Guerrini, A., & Tedesco, D. E. (2023). Restoring activity of milk thistle (Silybum marianum L.) on serum biochemical parameters, oxidative status, immunity, and performance in poultry and other animal species, poisoned by mycotoxins: A review. Animals, 13, 330.
Guo, L., Yao, H., Chen, W., Wang, X., Ye, P., Xu, Z., Zhang, S., & Wu, H. (2022). Natural products of medicinal plants: Biosynthesis and bioengineering in post-genomic era. Horticulture Research, 9, uhac223.
Gutyj, B. V., Martyshuk, T. V., Parchenko, V. V., Kaplaushenko, A. H., Bushueva, I. V., Hariv, I. I., Bilash, Y. P., Brygadyrenko, V. V., Turko, Y. I., & Radzykhovskyi, M. L. (2022). Effect of liposomal drug based on interferon and extract from Silybum marianum on antioxidative status of bulls against the background of contamination of fodders by cadmium and plumbum. Regulatory Mechanisms in Biosystems, 13(4), 419–425.
Gutyj, B. V., Ostapyuk, A. Y., Sobolev, O. I., Vishchur, V. J., Gubash, O. P., Kurtyak, B. M., Kovalskyi, Y. V., Darmohray, L. M., Hunchak, A. V., Tsisaryk, O. Y., Shcherbatyy, A. R., Farionik, T. V., Savchuk, L. B., Palyadichuk, O. R., & Hrymak, K. (2019). Cadmium burden impact on morphological and biochemical blood indicators of poultry. Ukrainian Journal of Ecology, 9(1), 235–239.
Gutyj, B., Stybel, V., Darmohray, L., Lavryshyn, Y., Turko, I., Hachak, Y., Shcherbatyy, A., Bushueva, I., Parchenko, V., Kaplaushenko, A., & Krushelnytska, O. (2017). Prooxidant-antioxidant balance in the organism of bulls (young cattle) after using cadmium load. Ukrainian Journal of Ecology, 7(4), 589–596.
Heidarian, E., & Nouri, A. (2021). Hepatoprotective effects of silymarin against diclofenac-induced liver toxicity in male rats based on biochemical parameters and histological study. Archives of Physiology and Biochemistry, 127(2), 112–118.
Islam, A., Mishra, A., Siddiqui, M. A., & Siddiquie, S. (2021). Recapitulation of evidence of phytochemical, pharmacokinetic and biomedical application of silybin. Drug Research (Stuttgart), 71(9), 489–503.
Javed, S., Ahsan, W., & Kohli, K. (2018). Pharmacological influences of natural products as bioenhancers of silymarin against carbon tetrachloride-induced hepatotoxicity in rats. Clinical Phytoscience, 4, 18.
Jiang, G., Sun, C., Wang, X., Mei, J., Li, C., Zhan, H., Liao, Y., Zhu, Y., & Mao, J. (2022). Hepatoprotective mechanism of Silybum marianum on nonalcoholic fatty liver disease based on network pharmacology and experimental verification. Bioengineered, 13(3), 5216–5235.
Juráňová, J., Aury-Landas, J., Boumediene, K., Baugé, C., Biedermann, D., Ulrichová, J., & Franková, J. (2019). Modulation of skin inflammatory response by active components of silymarin. Molecules, 24(1), 123.
Juretić, N., Sepúlveda, R., D'Espessailles, A., Vera, D. B., Cadagan, C., de Miguel, M., González-Mañán, D., & Tapia, G. (2021). Dietary alpha- and gamma-tocopherol (1:5 ratio) supplementation attenuates adipose tissue expansion, hepatic steatosis, and expression of inflammatory markers in a high-fat-diet-fed murine model. Nutrition, 85, 111139.
Karpenko, Y., Hunchak, Y., Gutyj, B., Hunchak, A., Parchenko, M., & Parchenko, V. (2022). Advanced research for physico-chemical properties and parameters of toxicity piperazinium 2-((5-(furan-2-YL)-4-phenyl-4H-1,2,4-triazol-3-yl) thio)acetate. ScienceRise: Pharmaceutical Science, 36, 18–25.
Khaleghipour, B., Khosravinia, H., Toghiyani, M., & Azarfar, A. (2019). Effects of silymarin on productive performance, liver function and serum biochemical profile in broiler Japanese quail challenged with dietary aflatoxins. Italian Journal of Animal Science, 18(1), 564–573.
Khazaei, R., Seidavi, A., & Bouyeh, M. (2022). A review on the mechanisms of the effect of silymarin in milk thistle (Silybum marianum) on some laboratory animals. Veterinary Medicine and Science, 8(1), 289–301.
Li, H.-Y., Gan, R.-Y., Shang, A., Mao, Q.-Q., Sun, Q.-C., Wu, D.-T., Geng, F., He, X.-Q., & Li, H.-B. (2021). Plant-based foods and their bioactive compounds on fatty liver disease: Effects, mechanisms, and clinical application. Oxidative Medicine and Cellular Longevity, 2021, 6621644.
Liang, M., Guo, M., Saw, P. E., & Yao, Y. (2022). Fully natural lecithin encapsulated nano-resveratrol for anti-cancer therapy. International Journal of Nanomedicine, 17, 2069–2078.
Lieshchova, M. A., & Brygadyrenko, V. V. (2023). Effect of Echinacea purpurea and Silybum marianum seeds on the body of rats with an excessive fat diet. Biosystems Diversity, 31(1), 90–99.
Lou-Bonafonte, J. M., Martínez-Beamonte, R., Sanclemente, T., Surra, J. C., Herrera-Marcos, L. V., Sanchez-Marco, J., Arnal, C., & Osada, J. (2018). Current insights into the biological action of squalene. Molecular Nutrition and Food Research, 62(15), 1800136.
Lozano-Grande, M. A., Gorinstein, S., Espitia-Rangel, E., Dávila-Ortiz, G., & Martínez-Ayala, A. L. (2018). Plant sources, extraction methods and uses of squalene. International Journal of Agronomy, 2018, 1829160.
Martyshuk, T. V., Gutyj, B. V., Vishchur, O. I., & Todoriuk, V. B. (2019). Biochemical indices of piglets blood under the action of feed additive “Butaselmevit-plus”. Ukrainian Journal of Veterinary and Agricultural Sciences, 2(2), 27–30.
Martyshuk, T. V., Gutyj, B. V., Zhelavskyi, M. M., Midyk. S. V., Fedorchenko, A. M., Todoriuk, V. B., Nahirniak, T. B., Kisera, Y. V., Sus, H. V., Chemerys, V. A., Levkivska, N. D., & Iglitskej, I. I. (2020). Effect of Butaselmevit-Plus on the immune system of piglets during and after weaning. Ukrainian Journal of Ecology, 10(2), 347–352.
Martyshuk, T., Gutyj, B., Vyshchur, O., Paterega, I., Kushnir, V., Bigdan, O., & Tkachenko, N. (2022). Study of acute and chronic toxicity of “Butaselmevit” on laboratory animals. Archives of Pharmacy Practice, 13(3), 70–75.
Mengesha, T., Gnanasekaran, N., & Mehare, T. (2021). Hepatoprotective effect of silymarin on fructose induced nonalcoholic fatty liver disease in male albino wistar rats. BMC Complementary Medicine and Therapies, 21(1), 104.
Molaei, S., Amiri, F., Salimi, R., Ferdowsi, S., & Bahadori, M. (2022). Therapeutic effects of mesenchymal stem cells-conditioned medium derived from suspension cultivation or silymarin on liver failure mice. Molecular Biology Reports, 49, 10315–10325.
Nehmi, V. A., Murata, G. M., Moraes, R. C. M., Lima, G. C. A., De Miranda, D. A., Radloff, K., Costa, R. G. F., Jesus, J. C. R., De Freitas, J. A., Viana, N. I., Pimenta, R., Leite, K. R. M., Otoch, J. P., & Pessoa, A. F. M. (2021). A novel supplement with yeast β-glucan, prebiotic, minerals and Silybum marianum synergistically modulates metabolic and inflammatory pathways and improves steatosis in obese mice. Journal of Integrative Medicine, 19(5), 439–450.
Nehmi-Filho, V., Santamarina, A. B., de Freitas, J. A., Trarbach, E. B., de Oliveira, D. R., Palace-Berl, F., de Souza, E., de Miranda, D. A., Escamilla-Garcia, A., Otoch, J. P., & Pessoa, A. F. M. (2023). Novel nutraceutical supplements with yeast β-glucan, prebiotics, minerals, and Silybum marianum (silymarin) ameliorate obesity-related metabolic and clinical parameters: A double-blind randomized trial. Frontiers in Endocrinology, 13, 1089938.
Ravichandran, V., Lee, M., Cao, N. T. G., & Shim, M. S. (2021). Polysorbate-based drug formulations for brain-targeted drug delivery and anticancer therapy. Applied Sciences, 11(19), 9336.
Sakamoto, M. I., Murakami, A. E., Fernandes, A. M., Ospina-Rojas, I. C., Nunes, K. C., & Hirata, A. K. (2018). Performance and serum biochemical profile of Japanese quail supplemented with silymarin and contaminated with aflatoxin B1. Poultry Science, 97(1), 159–166.
Salmerón-Manzano, E., Garrido-Cardenas, J. A., & Manzano-Agugliaro, F. (2020). Worldwide research trends on medicinal plants. International Journal of Environmental Research and Public Health, 17(10), 3376.
Santamarina, A. B., Moraes, R. C. M., Nehmi, V., Murata, G. M., de Freitas, J. A., de Miranda, D. A., Cerqueira, A. R. A., Costa, S. K. P., Ferreira, A. F. F., Britto, L. R., de Camargo, J. A., de Oliveira, R. D., de Jesus, F. N., Otoch, J. P., & Pessoa, A. F. M. (2022). The symbiotic effect of a new nutraceutical with yeast β-glucan, prebiotics, minerals, and Silybum marianum (silymarin) for recovering metabolic homeostasis via Pgc-1α, Il-6, and Il-10 gene expression in a type-2 diabetes obesity model. Antioxidants, 11(3), 447.
Shahin, S., Mansour, A., Abdel-Rahim, M., El-Dahhar, A., El Basuini, M., & Elhetawy, A. (2023). Silymarin, Silybum marianum, supplemented weaning diet boosted survival, growth, antioxidant status, and fatty acids profile of seabass, dicentrarchus labrax. Annals of Animal Science, 23(1), 253–264.
Stoev, S., Mircheva, T., Denev, S., Chobanova, S., & Ivanov, V. (2021). The protective effect of silymarin against ochratoxin a induced histopathological and biochemical changes in chicks. Journal of Advanced Veterinary Research, 11(1), 1–8.
Tajmohammadi, A., Razavi, B. M., & Hosseinzadeh, H. (2018). Silybum marianum (milk thistle) and its main constituent, silymarin, as a potential therapeutic plant in metabolic syndrome: a review. Phytotherapy Research, 32(10), 1933–1949.
Tedesco, D. E., & Guerrini, A. (2022). Use of milk thistle in farm and companion animals: A review. Planta Medica, 89(6), 584–607.
Tvrdý, V., Pourová, J., Jirkovský, E., Křen, V., Valentová, K., & Mladěnka, P. (2021). Systematic review of pharmacokinetics and potential pharmacokinetic interactions of flavonolignans from silymarin. Medicinal Research Reviews, 41(4), 2195–2246.
Ungurianu, A., Zanfirescu, A., Nițulescu, G., & Margină, D. (2021). Vitamin E beyond its antioxidant label. Antioxidants, 10(5), 634.
Viñado, A., Castillejos, L., & Barroeta, A. C. (2019). Soybean lecithin high in free fatty acids for broiler chicken diets: impact on performance, fatty acid digestibility and saturation degree of adipose tissue. Animals, 9(10), 802.
Vivchar, R. Y., & Lapovets, L. Y. (2018). Effect of lecethin usage on changes of cytokin levels in patients with multiple sclerosis. Visnyk Problem Biolohiji і Medycyny, 4(1), 57–60.
Vlizlo, V. V., Prystupa, O. I., Slivinska, L. G., Lukashchuk, B. O., Hu, S., Gutyj, B. V., Maksymovych, I. A., Shcherbatyy, A. R., Lychuk, M. G., Chernushkin, B. O., Leno, M. I., Rusyn, V. I., Drach, M. P., Fedorovych, V. L., Zinko, H. O., & Yaremchuk, V. Y. (2021). Functional state of the liver in cows with fatty liver disease. Ukrainian Journal of Ecology, 11(3), 167–173.
Vudmaska, I., Petrukh, I., Sachko, S., Vlizlo, V., Kosenko, Y., Kozak, M., & Petruk, A. (2021). Using hop cones, vitamin E, methionine, choline and carnitine for treatment of subclinical ketosis in transition dairy cows. Advances in Animal and Veterinary Sciences, 9(1), 55–62.
Wadhwa, K., Pahwa, R., Kumar, M., Kumar, S., Sharma, P. C., Singh, G., Verma, R., Mittal, V., Singh, I., Kaushik, D., & Jeandet, P. (2022). Mechanistic insights into the pharmacological significance of silymarin. Molecules, 27(16), 5327.
Wagner, K.-H., Shiels, R. G., Lang, C. A., Khoei, N. S., & Bulmer, A. C. (2018). Diagnostic criteria and contributors to Gilbert’s syndrome. Critical Reviews in Clinical Laboratory Sciences, 55(2), 129–139.
Wahyuni, L., Wirjosentono, B., & Tamrin, T. (2020). Effect of surfactant tween 80 (polyoxyethylene sorbitan mono oleate) addition on viscosity and activation energy on making asphalt emulsion. Journal of Chemical Natural Resources, 2(2), 113–120.
Wang, X., Zhang, Z., & Wu, S.-C. (2020). Health benefits of Silybum marianum: Phytochemistry, pharmacology, and applications. Journal of Agricultural and Food Chemistry, 68(42), 11644–11664.
Yu, Z., Wu, F., Tian, J., Guo, X., & An, R. (2018). Protective effects of compound ammonium glycyrrhizin, L-arginine, silymarin and glucurolactone against liver damage induced by ochratoxin A in primary chicken hepatocytes. Molecular Medicine Reports, 18(3), 2551–2560.
Zelenina, O., Vlizlo, V., Kozak, M., Ostapiv, D., Samaryk, V., Dron, I., Stetsko, T., Skrypka, M., Tomchuk, V., Danchuk, O., & Levchenko, A. (2022). Antimicrobial activity of the PEGylated antibiotic enrofloxacin and its functional and structural effect on the liver in rats. Journal of Applied Pharmaceutical Science, 12(6), 68–75.
Zhang, C., Shao, Q., Liu, M., Wang, X., Loor, J. J., Jiang, Q., Cuan, S., Li, X., Wang, J., Li, Y., He, L., Huang, Y., Liu, G., & Lei, L. (2023). Liver fibrosis is a common pathological change in the liver of dairy cows with fatty liver. Journal of Dairy Science, 106(4), 2700–2715.
Zhang, R., Xu, D., Zhang, Y., Wang, R., Yang, N., Lou, Y., Zhao, H., Aa, J., Wang, G., & Xie, Y. (2021). Silybin restored CYP3A expression through the sirtuin 2/nuclear factor κ-B pathway in mouse nonalcoholic fatty liver disease. Drug Metabolism and Disposition, 49(9), 770–779.
Zhu, S. Y., Jiang, N., Yang, J., Tu, J., Zhou, Y., Xiao, X., & Dong, Y. (2018). Silybum marianum oil attenuates hepatic steatosis and oxidative stress in high fat diet-fed mice. Biomedicine and Pharmacotherapy, 100, 191–197.
Published
2023-08-23
How to Cite
Vlizlo, V., Prystupa, O., Slivinska, L., Gutyj, B., Maksymovych, I., Shcherbatyy, A., Lychuk, M., Partyka, U., Chernushkin, B., Rusyn, V., Leno, M., & Leskiv, K. (2023). Treatment of animals with fatty liver disease using a drug based on the seeds of Silybum marianum . Regulatory Mechanisms in Biosystems, 14(3), 424-431. https://doi.org/10.15421/10.15421/022362