Dietary silymarin effectively alleviated fumonisin B1-induced hepatopancreatic steatosis and fibrosis in juvenile grass carp (Ctenopharyngodon idella)

Abstract

Fumonisin B1 (FB1) is a widely distributed environmental pollutant that specifically targets the liver, leading to hepatotoxicity. However, there is limited reference data on the hepatotoxicity of FB1 in fish and the corresponding mitigation measures. Silymarin (SIL) is a safe natural plant extract that exhibits hepatoprotective effects. This study aimed to evaluate the influence of supplemented SIL in the diet on FB1-induced hepatopancreatic injury in grass carp (Ctenopharyngodon idella). The grass carp (10.92 ± 0.01 g, n = 720) were divided into four groups at random: controls, FB1 (4 mg/kg), SIL (60 mg/kg) and FB1 + SIL group for 30 days. The results indicated that FB1 induced growth inhibition (P < 0.05), hepatopancreatic damage, steatosis and fibrosis. Dietary SIL alleviated growth inhibition, hepatopancreatic steatosis, accompanied by an increase in the expression levels of molecules related to lipid droplet and mitochondrial interaction (P < 0.01), as well as the β-oxidation of fatty acids (P < 0.05). Reduced steatosis mitigated oxidative damage and endoplasmic reticulum stress (P < 0.05). These changes further relieved the activation of apoptosis (P  < 0.01), which further alleviated the activation of the toll-like receptor 4/NF-kappa B pathway (P < 0.01) and transforming growth factor/SMAD family member pathway (P < 0.05), ultimately mitigating hepatopancreatic fibrosis. These results suggested that dietary supplementation with SIL effectively mitigated FB1-induced hepatopancreatic damage. Thus, this study provided new insights into strategies for mitigating FB1-induced hepatotoxicity.

Keywords

Environmental pollutant / Mycotoxin / Novel feed additive / Hepatotoxicity / Steatosis / Fibrosis

References

[1] Aghemo A, Alekseeva OP, Angelico F, Bakulin IG, Bakulina NV, Bordin D, Bueverov AO, Drapkina OM, Gillessen A, Kagarmanova EM, Korochanskaya NV, Kucheryavii UA, Lazebnik LB, Livzan MA, Maev IV, Martynov AI, Osipenko MF, Sas EI, Starodubova A, Uspensky YPet al. . Role of silymarin as antioxidant in clinical management of chronic liver diseases: a narrative review. Ann Med. 2022, 54: 1548-1560.

[2] Ahmadifar E, Moghadam MS, Dawood MAO, Hoseinifar SH. Lactobacillus fermentum and/or ferulic acid improved the immune responses, antioxidative defence and resistance against Aeromonas hydrophila in common carp (Cyprinus carpio) fingerlings. Fish Shellfish Immunol. 2019, 94: 916-923.

[3] Ahmadifar E, Sheikhzadeh N, Roshanaei K, Dargahi N, Faggio C. Can dietary ginger (Zingiber officinale) alter biochemical and immunological parameters and gene expression related to growth, immunity and antioxidant system in zebrafish (Danio rerio)?. Aquaculture. 2019, 507: 341-348.

[4] Ahmadifar E, Hoseinifar SH, Adineh H, Moghadam MS, Dawood MaJaOaS. Assessing the impact of purslane (Portulaca oleracea L.) on growth performance, anti-oxidative, and immune activities in grass carp (Ctenopharyngodon idella). Ann Anim Sci. 2020, 20: 1427-1440.

[5] Ahmadifar E, Pourmohammadi Fallah H, Yousefi M, Dawood MA, Hoseinifar SH, Adineh H, Yilmaz S, Paolucci M, Doan HV. The gene regulatory roles of herbal extracts on the growth, immune system, and reproduction of fish. Animals. 2021, 11: 2167.

[6] Ahmadifar E, Yousefi M, Karimi M, Fadaei Raieni R, Dadar M, Yilmaz S, Dawood MA, Abdel-Latif HM. Benefits of dietary polyphenols and polyphenol-rich additives to aquatic animal health: an overview. Rev Fish Sci Aquac. 2021, 29478-511.

[7] AOAC. Official methods of analysis. 200518Washington, Association of Official Agricultural Chemists

[8] Armanini EH, Boiago MM, Cécere BGDO, Oliveira PV, Teixeira CJ, Strapazzon JV, Bottari NB, Silva AD, Fracasso M, Vendruscolo RG. Protective effects of silymarin in broiler feed contaminated by mycotoxins: growth performance, meat antioxidant status, and fatty acid profiles. Trop Anim Health Prod. 2021, 53: 442.

[9] Baldissera MD, Souza CF, Da Silva HNP, Henn AS, Duarte FA, Da Costa ST, Da Silva AS, Baldisserotto B. Diphenyl diselenide modulates splenic purinergic signaling in silver catfish fed diets contaminated with fumonisin B1: an attempt to improve immune and hemostatic responses. Comp Biochem Physiol C Toxicol Pharmacol. 2020, 227108624.

[10] Bello M. Structural basis of Nrf2 activation by flavonolignans from silymarin. J Mol Graph Model. 2023, 119108393.

[11] Braun M, Wink M. Exposure, occurrence, and chemistry of fumonisins and their cryptic derivatives. Compr Rev Food Sci Food Saf. 2018, 17769-791.

[12] Cao SS, Kaufman RJ. Endoplasmic reticulum stress and oxidative stress in cell fate decision and human disease. Antioxid Redox Signal. 2014, 21: 396-413.

[13] Cao C, Zhu X, Li X, Ouyang H, Wang K, Li X. Assessment of ionic homeostasis imbalance and cytochrome P450 system disturbance in mice during fumonisin B1 (FB1) exposure. Chemosphere. 2020, 251: 126393.

[14] Cao C, Xian R, Lin F, Li X, Li X, Qiang F, Li X. Fumonisin B1 induces hepatotoxicity in mice through the activation of oxidative stress, apoptosis and fibrosis. Chemosphere. 2022, 296133910.

[15] Chen J, Wen J, Tang Y, Shi J, Mu G, Yan R, Cai J. Research progress on fumonisin B1 contamination and toxicity: a review. Molecules. 2021, 265238.

[16] Chen D, Jiang W, Wu P, Liu Y, Ren H, Jin X, Zhou X, Feng L. Mechanism of fumonisin B1 on growth performance and intestinal structural integrity of juvenile grass carp (Ctenopharyngodon idella). Anim Nutr. 2025, 21193-206.

[17] Cho JH, Lee JS, Kim HG, Lee HW, Fang Z, Kwon HH, Kim DW, Lee CM, Jeong JW. Ethyl acetate fraction of Amomum villosum var. xanthioides attenuates hepatic endoplasmic reticulum stress-induced non-alcoholic steatohepatitis via improvement of antioxidant capacities. Antioxidants. 2021, 10: 998.

[18] Dewidar B, Meyer C, Dooley S, Meindl-Beinker AN. Tgf-β in hepatic stellate cell activation and liver fibrogenesis—updated 2019. Cells. 2019, 81419.

[19] Diao L, Auger C, Konoeda H, Sadri AR, Amini-Nik S, Jeschke MG. Hepatic steatosis associated with decreased β-oxidation and mitochondrial function contributes to cell damage in obese mice after thermal injury. Cell Death Dis. 2018, 9530.

[20] Dichtel LE, Cordoba Chacon J, Kineman RD. Growth hormone and insulin-like growth factor 1 regulation of nonalcoholic fatty liver disease. J Clin Endocrinol Metab. 2022, 1071812-1824.

[21] El-Kot SM, Wanas W, Hafez AM, Mahmoud NA, Tolba AM, Younis AH, Sayed GE, Abdelwahab HE. Effect of silymarin on the relative gene expressions of some inflammatory cytokines in the liver of CCl₄-intoxicated male rats. Sci Rep. 2023, 13: 15245.

[22] Enkler L, Spang A. Functional interplay of lipid droplets and mitochondria. FEBS Lett. 2024, 5981235-1251.

[23] Faienza MF, Brunetti G, Ventura A, D’aniello M, Pepe T, Giordano P, Monteduro M, Cavallo L. Nonalcoholic fatty liver disease in prepubertal children born small for gestational age: influence of rapid weight catch-up growth. Horm Res Paediatr. 2013, 79: 103-109.

[24] Fromenty B, Roden M. Mitochondrial alterations in fatty liver diseases. J Hepatol. 2023, 78: 415-429.

[25] Gao Z, Luo K, Peng J, Zhu Q, Liu C, Wang X, Li S, Zhang H. The natural occurrence, toxicity mechanisms and management strategies of fumonisin B1: a review. Environ Pollut. 2023, 320: 121065.

[26] García-Muñoz AM, Victoria-Montesinos D, Ballester P, Cerdá B, Zafrilla P. A descriptive review of the antioxidant effects and mechanisms of action of berberine and silymarin. Molecules. 2024, 294576.

[27] Gbore F, Adewole A, Oginni O, Oguntolu M, Bada A, Akele O. Growth performance, haematology and serum biochemistry of african catfish (Clarias gariepinus) fingerlings fed graded levels of dietary fumonisin b 1. Mycotoxin Res. 2010, 26221-227.

[28] Gonçalves R, Schatzmayr D, Hofstetter U, Santos G. Occurrence of mycotoxins in aquaculture: preliminary overview of asian and european plant ingredients and finished feeds. World Mycotoxin J. 2017, 10: 183-194.

[29] Gonçalves R, Hofstetter U, Schatzmayr D, Jenkins T. Mycotoxins in southeast asian aquaculture: plant-based meals and finished feeds. World Mycotoxin J. 2018, 11: 265-276.

[30] He X, Gao J, Hou H, Qi Z, Chen H, Zhang XX. Inhibition of mitochondrial fatty acid oxidation contributes to development of nonalcoholic fatty liver disease induced by environmental cadmium exposure. Environ Sci Technol. 2019, 53: 13992-14000.

[31] Ignat SR, Dinescu S, Hermenean A, Costache M. Cellular interplay as a consequence of inflammatory signals leading to liver fibrosis development. Cells. 2020, 9: 461.

[32] Kim SH, Singh MP, Sharma C, Kang SC. Fumonisin B1 actuates oxidative stress-associated colonic damage via apoptosis and autophagy activation in murine model. J Biochem Mol Toxicol. 2018, 22e22161.

[33] Kiruthiga PV, Shafreen RB, Pandian SK, Devi KP. Silymarin protection against major reactive oxygen species released by environmental toxins: exogenous H₂O₂ exposure in erythrocytes. Basic Clin Pharmacol Toxicol. 2007, 100414-419.

[34] Kracizy RO, Brazão CC, De Marco Viott A, Ribeiro K, Koppenol A, Dos Santos AMV, Ballester ELC. Evaluation of aflatoxin and fumonisin in the diet of Pacific white shrimp (Litopenaeus vannamei) on their performance and health. Aquaculture. 2021, 544: 737051.

[35] Liu Q, Wang Q, Xu C, Shao W, Zhang C, Liu H, Jiang Z, Gu A. Organochloride pesticides impaired mitochondrial function in hepatocytes and aggravated disorders of fatty acid metabolism. Sci Rep. 2017, 746339.

[36] Liu Q, Zou X, Zhao M, Guan Q, Xuan Z, Liu L, Gao Z. Integrated transcriptome and metabolome analysis of liver reveals unsynchronized growth mechanisms in blunt-snout bream (Megalobrama amblycephala). BMC Genom. 2025, 2630.

[37] Lumlertdacha S, Lovell RT, Shelby RA, Lenz SD, Kemppainen BW. Growth, hematology, and histopathology of channel catfish, Ictalurus punctatus, fed toxins from Fusarium moniliforme. Aquaculture. 1995, 130201-218.

[38] Ma Y, Lee G, Heo S, Roh Y. Oxidative stress is a key modulator in the development of nonalcoholic fatty liver disease. Antioxidants. 2022, 11: 91.

[39] Murao A, Aziz M, Wang H, Brenner M, Wang P. Release mechanisms of major damps. Apoptosis. 2021, 26152-162.

[40] Nagata S. Apoptosis by death factor. Cell. 1997, 88: 355-365.

[41] NRC. Nutrient requirements of fish and shrimp. 2011, Washington, National Academic Press

[42] Ong G, Logue SE. Unfolding the interactions between endoplasmic reticulum stress and oxidative stress. Antioxidants. 2023, 12: 981.

[43] Oyadomari S, Mori M. Roles of chop/gadd153 in endoplasmic reticulum stress. Cell Death Differ. 2004, 11381-389.

[44] Salomone F, Barbagallo I, Godos J, Lembo V, Currenti W, Cinà D, Avola R, D’orazio N, Morisco F, Galvano F, Li Volti G. Silibinin restores NAD⁺ levels and induces the sirt1/ampk pathway in non-alcoholic fatty liver. Nutrients. 2017, 9: 1086.

[45] Singh MP, Kang SC. Endoplasmic reticulum stress-mediated autophagy activation attenuates fumonisin B1 induced hepatotoxicity in vitro and in vivo. Food Chem Toxicol. 2017, 110371-382.

[46] Sumana SL, Xue T, Hu H, Abdullateef MM, Shui Y, Ayana GU, Kayiira JC, Zhang C, Samwel BJ, Zhu J. Medicinal plants as ecological solutions for fish growth and immunostimulatory effects in aquaculture. Aquaculture. 2025, 2025: 9778623.

[47] Surai PF, Surai A, Earle-Payne K. Silymarin and inflammation: food for thoughts. Antioxidants. 2024, 1398.

[48] Thapaliya S, Wree A, Povero D, Inzaugarat ME, Berk M, Dixon L, Papouchado BG, Feldstein AE. Caspase 3 inactivation protects against hepatic cell death and ameliorates fibrogenesis in a diet-induced NASH model. Dig Dis Sci. 2014, 591197-1206.

[49] Trappoliere M, Caligiuri A, Schmid M, Bertolani C, Failli P, Vizzutti F, Novo E, Di Manzano C, Marra F, Loguercio C, Pinzani M. Silybin, a component of sylimarin, exerts anti-inflammatory and anti-fibrogenic effects on human hepatic stellate cells. J Hepatol. 2009, 50: 1102-1111.

[50] Wang XQ, Chang YH, Wang XC, Liu RQ, Yang SJ, Hu ZY, Jiang FW, Chen M-S, Wang JX, Liu S. Sirt1 regulates fumonisin B1-induced LMH cell panoptosis and antagonism of lycopene. J Agric Food Chem. 2025, 73: 4923-4935.

[51] Wei X-Q, Wu W-D, Zhou Y, Jiang S-Y, Liu L, Kuang L, Tang FW, Feng LJA, Wang JX. Dietary silymarin supplementation enhanced growth performance and improved intestinal apical junctional complex on juvenile grass carp (Ctenopharyngodon idella). Aquaculture. 2020, 525: 735311.

[52] Wigraiboon S, Panchan R, Luang-In V, Ounjit W, Panase P, Sookying S, Sutthi N. Effects of dietary tuber ethanolic extract of nut grass (Cyperus rotundus L.) on growth, immune response, and disease resistance in Nile tilapia (Oreochromis niloticus). Animals. 2024, 14503.

[53] Yayeh T, Jeong HR, Park YS, Moon S, Sur B, Yoo HS, Oh S. Fumonisin B1-induced toxicity was not exacerbated in glutathione peroxidase-1/catalase double knock out mice. Biomol Ther. 2021, 2952-57.

[54] Zhang H, Jiang W, Wu P, Liu Y, Ren H, Jin X, Zhou X, Feng L. Revealing high-efficiency natural mycotoxin antidotes in zebrafish model screening against zearalenone-induced toxicity. Engineering. 2024, 42: 196-212.

[55] Zhao J, Feng L, Liu Y, Jiang W, Wu P, Jiang J, Zhang Y, Zhou X. Effect of dietary isoleucine on the immunity, antioxidant status, tight junctions and microflora in the intestine of juvenile jian carp (Cyprinus carpio var. Jian). Fish Shellfish Immunol. 2014, 41: 663-673.

[56] Zhao Y, Yan Y, Zhao Z, Li S, Yin J. The dynamic changes of endoplasmic reticulum stress pathway markers grp78 and chop in the hippocampus of diabetic mice. Brain Res Bull. 2015, 111: 27-35.

[57] Zhao P, Zhang L, Feng L, Jiang WD, Wu P, Liu Y, Ren HM, Jin XW, Zhou XQ. Novel perspective on mechanism in muscle growth inhibited by ochratoxin a associated with ferroptosis: model of juvenile grass carp (Ctenopharyngodon idella) in vivo and in vitro trials. J Agric Food Chem. 2024, 72: 4977-4990.

[58] Zheng C, Wang L, Zou T, Lian S, Luo J, Lu Y, Hao H, Xu Y, Xiang Y, Zhang X, Xu G, Zou X, Jiang R. Ileitis promotes masld progression via bile acid modulation and enhanced TGR5 signaling in ileal CD8+ T cells. J Hepatol. 2024, 80: 764-777.

[59] Zhu L, Li J, Yang S, Deng X, Wang Z, Cao C. Fumonisin B1 induces endoplasmic reticulum damage and inflammation by activating the NXR response and disrupting the normal CYP450 system, leading to liver damage in juvenile quail. J Food Sci. 2024, 89: 5967-5979.

RIGHTS & PERMISSIONS

The Author(s)