Reactive oxygen species (ROS) burst as a primary driver of high-carbohydrate diet-induced metabolic liver disease in carnivorous fish

Abstract

High-carbohydrate diets (HCD) induce highly consistent metabolic liver disease in carnivorous fish, commonly characterized by redox imbalance and dysregulation of glycolipid metabolism. However, the role of elevated reactive oxygen species (ROS) under oxidative stress in this pathological process remains inadequately elucidated. This study investigated the mechanism by which HCD-induced ROS burst drives liver damage in the carnivorous fish, largemouth bass (Micropterus salmoides). Through integrated physiological, biochemical, and multi-omics analyses, we found that HCD significantly elevated the liver ROS levels, inducing oxidative stress (increased malondialdehyde and suppressed antioxidant enzyme activities), mitochondrial dysfunction (reduced membrane potential, mtDNA damage, and ultrastructural disruption), and inhibition of the PINK1/Parkin-mediated mitophagy pathway. To scavenge excess ROS, the mitochondria-targeted antioxidant Mito-TEMPO was employed for intervention. The results demonstrated that the aforementioned pathological phenotypes were significantly ameliorated, and mitophagy was restored, confirming ROS burst as a core factor exacerbating HCD-induced liver damage. Furthermore, integrated transcriptomic and metabolomic analysis revealed that ROS burst and its scavenging can remodel the sphingolipid and amino acid metabolic networks, with gba2 and chac1 identified as key hub genes. These findings elucidate the central role of ROS in HCD-induced metabolic liver disease in carnivorous fish, providing insights for antioxidant-based intervention strategies against metabolic disorders in aquaculture.

Keywords

ROS burst / High-carbohydrate diets / Metabolic disorder / PINK1/Parkin pathway

References

[1] Bourganou MV, Chondrogianni ME, Kyrou I, Flessa CM, Chatzigeorgiou A, Oikonomou E, Lambadiari V, Randeva HS, Kassi E. Unraveling metabolic dysfunction-associated steatotic liver disease through the use of omics technologies. Int J Mol Sci. 2025, 26. 1589

[2] Buzzetti E, Pinzani M, Tsochatzis EA. The multiple-hit pathogenesis of non-alcoholic fatty liver disease (NAFLD). Metabolism. 2016, 65: 1038-1048.

[3] Caputo V, Tarantino G, Santini SJ, Fracassi G, Balsano C. The role of epigenetic control of mitochondrial (Dys)function in MASLD onset and progression. Nutrients. 2023, 15: 4757.

[4] Chang B, Su Y, Li T, Zheng Y, Yang R, Lu H, Wang H, Ding Y. Mito-TEMPO ameliorates sodium palmitate induced ferroptosis in MIN6 cells through PINK1/Parkin-mediated mitophagy. Biomed Environ Sci. 2024, 371128-1141

[5] Chen P, Yao L, Yuan M, Wang Z, Zhang Q, Jiang Y, Li L. Mitochondrial dysfunction: a promising therapeutic target for liver diseases. Genes Dis. 2024, 11. 101115

[6] Deng Y, Zhang W, Yang Z, Kong Q, Liu P, Liao H, Cui Z, Tang H. Dietary lactobacillus plantarum can alleviate high starch diet-induced liver lipid deposition, tissue damage and oxidative stress in largemouth bass (Micropterus salmoides). Aquacult Rep. 2024, 35. 101955

[7] Devasia AG, Ramasamy A, Leo CH. Current therapeutic landscape for metabolic dysfunction-associated steatohepatitis. Int J Mol Sci. 2025, 26. 1778

[8] Dong Y, Yu M, Wu Y, Xia T, Wang L, Song K, Zhang C, Lu K, Rahimnejad S. Hydroxytyrosol promotes the mitochondrial function through activating mitophagy. Antioxidants. 2022, 11: 893.

[9] Dong Y, Zhuang X, Wang Y, Tan J, Feng D, Li M, Zhong Q, Song Z, Shen HM, Fang EF, Lu J. Chemical mitophagy modulators: drug development strategies and novel regulatory mechanisms. Pharmacol Res. 2023, 194. 106835

[10] Dungubat E, Fujikura K, Kuroda M, Fukusato T, Takahashi Y. Food nutrients and bioactive compounds for managing metabolic dysfunction-associated steatotic liver disease: a comprehensive review. Nutrients. 2025, 172211.

[11] El-Sehrawy AAMA, Mohammed AN, Gupta J, Mohammed JS, Roopashree R, Kashyap A, Janney JB, Sahoo S, Al-Hasnaawei S, Nasr YM. Combating oxidative stress in non-alcoholic fatty liver disease: from mechanisms to therapeutic strategies. Pathol Res Pract. 2025, 272. 156053

[12] Enes P, Panserat S, Kaushik S, Oliva-Teles A. Nutritional regulation of hepatic glucose metabolism in fish. Fish Physiol Biochem. 2009, 35519-539.

[13] Fan Y, Li S, Yang X, Bai S, Tang M, Zhang X, Lu C, Ji C, Du G, Qin Y. Multi-omics approach characterizes the role of Bisphenol F in disrupting hepatic lipid metabolism. Environ Int. 2024, 187. 108690

[14] Feng Z, Zhong Y, He G, Sun H, Chen Y, Zhou W, Lin S. Yeast culture improved the growth performance, liver function, intestinal barrier and microbiota of juvenile largemouth bass (Micropterus salmoides) fed high-starch diet. Fish Shellfish Immunol. 2022, 120: 706-715.

[15] Ge Y, Zhang L, Chen W, Sun M, Liu W, Li X. Resveratrol modulates the redox response and bile acid metabolism to maintain the cholesterol homeostasis in fish Megalobrama amblycephala offered a high-carbohydrate diet. Antioxidants. 2023, 12. 121

[16] Geng H, Xu W, Wu L, Dong B, Han D, Liu H, Zhu X, Yang Y, Xie S, Jin J. Early programming effects of a hyperglucidic diet on carbohydrate and lipid metabolism in gibel carp (Carassius gibelio). Aquacult Rep. 2023, 33. 101838

[17] Huang X, Ouyang X, Wang L, Xue K, Yu H, Liao C, Chen F, Rong K, Zhang X. Longitudinal analysis of metabolic liver disease induced by high-starch diets in largemouth bass (Micropterus salmoides). Aquacult Int. 2025, 33: 333.

[18] Iorio R, Celenza G, Petricca S. Mitophagy: molecular mechanisms, new concepts on Parkin activation and the emerging role of AMPK/ULK1 axis. Cells. 2021, 11. 30

[19] Jiang H. AMPK: balancing mitochondrial quality and quantity through opposite regulation of mitophagy pathways. Mol Cell. 2024, 844261-4263.

[20] Jin S, Li Y, Xia T, Liu Y, Zhang S, Hu H, Chang Q, Yan M. Mechanisms and therapeutic implications of selective autophagy in nonalcoholic fatty liver disease. J Adv Res. 2025, 67: 317-329.

[21] Ju Y, Zhang Y, Tian X, Zhu N, Zheng Y, Qiao Y, Yang T, Niu B, Li X, Yu L, Liu Z, Wu Y, Zhi Y, Dong Y, Xu Q, Yang X, Wang X, Wang X, Deng H, Mao Yet al. . Protein S-glutathionylation confers cellular resistance to ferroptosis induced by glutathione depletion. Redox Biol. 2025, 83. 103660

[22] Körschen HG, Yildiz Y, Raju DN, Schonauer S, Bönigk W, Jansen V, Kremmer E, Kaupp UB, Wachten D. The non-lysosomal β-glucosidase GBA2 is a non-integral membrane-associated protein at the endoplasmic reticulum (ER) and Golgi. J Biol Chem. 2013, 288: 3381-3393.

[23] Kumar S, Enes P, Singha KP, Esmaeili N, Soengas JL, Panserat S. Kumar S. Nutrition and physiology of fish and shellfish: feed regulation, metabolism and digestion. Metabolism of carbohydrates. 20251Amsterdam, Elsevier349393

[24] Li L, Wang H, Zhao S, Zhao Y, Chen Y, Zhang J, Wang C, Sun N, Fan H. Paeoniflorin ameliorates lipopolysaccharide-induced acute liver injury by inhibiting oxidative stress and inflammation via SIRT1/FOXO1a/SOD2 signaling in rats. Phytother Res. 2022, 36: 2558-2571.

[25] Li D, Tian L, Nan P, Zhang J, Zheng Y, Jia X, Gong Y, Wu Z. CerS6 triggered by high glucose activating the TLR4/IKKβ pathway regulates ferroptosis of LO2 cells through mitochondrial oxidative stress. Mol Cell Endocrinol. 2023, 572. 111969

[26] Li R, Chen L, Limbu SM, Qian Y, Zhou W, Chen L, Luo Y, Qiao F, Zhang M, Du Z. High cholesterol intake remodels cholesterol turnover and energy homeostasis in Nile tilapia (Oreochromis niloticus). Mar Life Sci Technol. 2023, 5: 56-74.

[27] Li H, Zeng Y, Wang G, Zhang K, Gong W, Li Z, Tian J, Xia Y, Xie W, Xie J, Xie S, Yu E. Betaine improves appetite regulation and glucose-lipid metabolism in mandarin fish (Siniperca chuatsi) fed a high-carbohydrate-diet by regulating the AMPK/mTOR signaling. Heliyon. 2024, 10. e28423

[28] Li J, Ni Y, Zhang Y, Liu H. GBA3 promotes fatty acid oxidation and alleviates non-alcoholic fatty liver by increasing CPT2 transcription. Aging. 2024, 16: 4591-4608

[29] Lin L, Chen Z, Huang C, Wu Y, Huang L, Wang L, Ke S, Liu L. Mito-TEMPO, a mitochondria-targeted antioxidant, improves cognitive dysfunction due to hypoglycemia: an association with reduced pericyte loss and blood-brain barrier leakage. Mol Neurobiol. 2023, 60: 672-686.

[30] Liu J, Tang D, Kang R. Targeting GPX4 in ferroptosis and cancer: chemical strategies and challenges. Trends Pharmacol Sci. 2024, 45: 666-670.

[31] Liu S, Liu J, Wang Y, Deng F, Deng Z. Oxidative stress: signaling pathways, biological functions, and disease. MedComm. 2025, 6. e70268

[32] Pang X, Yang J, Xiang SA, Sun H, Fu SJ. Interspecific differences in growth, digestion, and feeding metabolism among freshwater fishes with different food habits. Front Mar Sci. 2024, 11. 1490406

[33] Paredes JF, Habte-Tsion M, Riche M, Mejri S, Bradshaw D, Chin LS, Perricone C, Wills PS. Effects of different carbohydrates on growth, hepatic glucose metabolism, and gut microbiome of Florida pompano (Trachinotus carolinus). Aquaculture. 2025, 600. 742237

[34] Radosavljevic T, Brankovic M, Samardzic J, Djuretić J, Vukicevic D, Vucevic D, Jakovljevic V. Altered mitochondrial function in MASLD: key features and promising therapeutic approaches. Antioxidants. 2024, 13: 906.

[35] Undamatla R, Fagunloye OG, Chen J, Edmunds LR, Murali A, Mills A, Xie B, Pangburn MM, Sipula I, Gibson G, St Croix C, Jurczak MJ. Reduced mitophagy is an early feature of NAFLD and liver-specific PARKIN knockout hastens the onset of steatosis, inflammation and fibrosis. Sci Rep. 2023, 13. 7575

[36] Vikas K, Paula E, Krishna PS, Noah E, Jose L, Stephane P. Nutrition and physiology of fish and shellfish: feed regulation, metabolism and digestion. 2025, Amsterdam, Elsevier

[37] Wang PF, Xie K, Cao YX, Zhang A. Hepatoprotective effect of mitochondria-targeted antioxidant Mito-TEMPO against lipopolysaccharide-induced liver injury in mouse. Mediators Inflamm. 2022, 2022. 6394199

[38] Xue M, Yao T, Xue M, Francis F, Qin Y, Jia M, Li J, Gu X. Mechanism analysis of metabolic fatty liver on largemouth bass (Micropterus salmoides) based on integrated lipidomics and proteomics. Metabolites. 2022, 121-21.

[39] Yang H, Ou-Yang K, He Y, Wang X, Wang L, Yang Q, Li D, Li L. Nitrite induces hepatic glucose and lipid metabolism disorders in zebrafish through mitochondrial dysfunction and ERs response. Aquat Toxicol. 2024, 273. 107015

[40] Yang J, Zhang J, Zhang L, Yang Z. Mitochondrial oxidative stress-associated mechanisms in the development of metabolic dysfunction-associated steatotic liver disease. Biocell. 2025, 49: 399-417.

[41] Yu H, Nie Y, Zhang B, Xue J, Xue K, Huang X, Zhang X. Creatine supplementation in largemouth bass (Micropterus salmoides) diets: improving intestinal health and alleviating enteritis. Fish Shellfish Immunol. 2025, 159. 110164

[42] Zhang Y, Liang XF, He S, Wang J, Li L, Zhang Z, Li J, Chen X, Li L, Alam MS. Metabolic responses of Chinese perch (Siniperca chuatsi) to different levels of dietary carbohydrate. Fish Physiol Biochem. 2021, 47: 1449-1465.

[43] Zhang Y, Chen Q, Fu X, Zhu S, Huang Q, Li C. Current advances in the regulatory effects of bioactive compounds from dietary resources on nonalcoholic fatty liver disease: role of autophagy. J Agric Food Chem. 2023, 71: 17554-17569.

[44] Zhang R, Yan Z, Zhong H, Luo R, Liu W, Xiong S, Liu Q, Liu M. Gut microbial metabolites in MASLD: implications of mitochondrial dysfunction in the pathogenesis and treatment. Hepatol Commun. 2024, 8. e0484

[45] Zhao W, Wei HL, Wang ZQ, He XS, Niu J. Effects of dietary carbohydrate levels on growth performance, body composition, antioxidant capacity, immunity, and liver morphology in Oncorhynchus mykiss under cage culture with flowing freshwater. Aquacult Nutr. 2022, 20227820017.

[46] Zhou M, Lv J, Chen X, Shi Y, Chao G, Zhang S. From gut to liver: exploring the crosstalk between gut-liver axis and oxidative stress in metabolic dysfunction-associated steatotic liver disease. Ann Hepatol. 2025, 30. 101777

[47] Zou J, Chen Z, Zheng F, Cao X, Zhao Y, Panserat S, Gao J, Wang Q. Glycogenic hepatopathy in a primitive teleost fish model: the inductive effect of high carbohydrate diet and the alleviating role of betaine. Mar Life Sci Technol. 2025.

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