Lutein mitigates high-fat-diet induced hepatic lipid accumulation and fibrosis by targeting Hmgcs1 in marine fish

Abstract

This study investigated the alleviating effects of dietary lutein (LU) against high-fat diet (HFD)-induced hepatic lipid accumulation and fibrosis in black seabream (Acanthopagrus schlegelii). A 10-week feeding trial demonstrated that while LU did not alter growth performance, it significantly mitigated HFD-induced visceral adiposity. LU supplementation markedly lowered serum triglycerides (TG), cholesterol (CHO), and low-density lipoprotein cholesterol (LDL-C) levels compared to the HFD group. Histological analysis confirmed reduced hepatic lipid droplet accumulation, accompanied by decreased malondialdehyde (MDA) and reactive oxygen species (ROS) levels, alongside enhanced antioxidant enzyme activities. Transcriptomic profiling revealed that LU downregulated genes involved in cholesterol biosynthesis and fibrosis, with 3-hydroxy-3-methylglutaryl-CoA synthase 1 (hmgcs1) identified as a key target. Molecular docking and cellular thermal shift assay results supported an interaction between LU and Hmgcs1. In the established black seabream hepatocytes, LU suppressed oleic acid- (OA)-induced cholesterol biosynthesis and fibrotic gene expression, whereas hmgcs1 knockdown enhanced and hmgcs1 overexpression attenuated these protective effects. Collectively, dietary LU reduced HFD-induced hepatic lipid accumulation, oxidative stress, and fibrotic responses in black seabream, at least partly through downregulation of hmgcs1-associated cholesterol biosynthesis.

Keywords

Acanthopagrus schlegelii / High-fat diet / Hmgcsl / Lutein / Liver injury

References

[1] Ai Q, Zhao J, Mai K, Xu W, Tan B, Ma H, Liufu Z. Optimal dietary lipid level for large yellow croaker (Pseudosciaena crocea) larvae. Aquac Nutr, 2008, 14: 515-522

[2] Akkewar A, Mishra K, Kamble M, Kumar S, Dey J, Sethi K. A mechanistic review on growing multiple therapeutic applications of lutein and its global market research. Phytother Res, 2024, 38: 3190-3217

[3] AOAC. Official methods of analysis, 2006, 18th edn, Arlington Association of Official Analytical Chemists

[4] Arguello G, Balboa E, Arrese M, Zanlungo S. Recent insights on the role of cholesterol in non-alcoholic fatty liver disease. Biochimica Et Biophysica Acta (BBA) - Molecular Basis of Disease, 2015, 1852: 1765-1778

[5] Aust O, Sies H, Stahl W, Polidori M. Analysis of lipophilic antioxidants in human serum and tissues: tocopherols and carotenoids. J Chromatogr A, 2001, 936: 83-93

[6] Bao Y, Zhu J, Yang Q, Wu L, Zhao W, Zhu T, Sun P, Zhou Q, Jin M. Dietary lutein mitigates high fat diet and lipopolysaccharide induced intestinal barrier dysfunction and immune inflammatory responses in juvenile black seabream (Acanthopagrus schlegelii). Fish Shellfish Immunol, 2026, 177 111609

[7] Besen K, Melim E, Da Cunha L, Favaretto E, Moreira M, Fabregat TEHP. Lutein as a natural carotenoid source: effect on growth, survival and skin pigmentation of goldfish juveniles (Carassius auratus). Aquac Res, 2019, 50: 2200-2206

[8] Boujard T, Gélineau A, Covès D, Corraze G, Dutto G, Gasset E, Kaushik S. Regulation of feed intake, growth, nutrient and energy utilisation in European sea bass (Dicentrarchus labrax) fed high fat diets. Aquaculture, 2004, 231: 529-545

[9] Boyd C, D’Abramo L, Glencross B, Huyben D, Juarez L, Lockwood G, McNevin A, Tacon A, Teletchea F, Tomasso J, Tucker C, Valenti W. Achieving sustainable aquaculture: historical and current perspectives and future needs and challenges. J World Aquac Soc, 2020, 51: 578-633

[10] Caldas H. NSDHL, an enzyme involved in cholesterol biosynthesis, traffics through the Golgi and accumulates on ER membranes and on the surface of lipid droplets. Hum Mol Genet, 2003, 12: 2981-2991

[11] Cheng J, Liu D, Zhao J, Li X, Yan Y, Wu Z, Wang H, Wang C. Lutein attenuates oxidative stress and inhibits lipid accumulation in free fatty acids-induced HepG2 cells by activating the AMPK pathway. J Funct Foods, 2019, 60 103445

[12] Chou B, Shiau S. Optimal dietary lipid level for growth of juvenile hybrid tilapia, Oreochromis niloticus × Oreochromis aureus. Aquaculture, 1996, 143: 185-195

[13] Feng R, Du W, Lui P, Zhang J, Liu Y. CAPN2 acts as an indicator of hepatitis B virus to induce hepatic fibrosis. J Cell Biochem, 2020, 121: 2428-2436

[14] Fuad NIN, Sekar M, Gan SH, Lum PT, Vaijanathappa J, Ravi S. Lutein: a comprehensive review on its chemical, biological activities and therapeutic potentials. Pharmacogn J, 2020, 12: 1769-1778

[15] Hu N, Chen C, Wang J, Huang J, Yao D, Li C. Atorvastatin ester regulates lipid metabolism in hyperlipidemia rats via the PPAR-signaling pathway and HMGCR expression in the liver. Int J Mol Sci, 2021, 22 11107

[16] Jafari R, Almqvist H, Axelsson H, Ignatushchenko M, Lundbäck T, Nordlund P, Molina D. The cellular thermal shift assay for evaluating drug target interactions in cells. Nat Protoc, 2014, 9: 2100-2122

[17] Jin M, Zhu T, Tocher D, Luo J, Shen Y, Li X, Pan T, Yuan Y, Betancor M, Jiao L, Sun P, Zhou Q. Dietary fenofibrate attenuated high-fat-diet-induced lipid accumulation and inflammation response partly through regulation of pparα and sirt1 in juvenile black seabream (Acanthopagrus schlegelii). Dev Comp Immunol, 2020, 109 103691

[18] Jokela H, Rantakari P, Lamminen T, Strauss L, Ola R, Mutka AL, Gylling H, Miettinen T, Pakarinen P, Sainio K, Poutanen M. Hydroxysteroid (17β) dehydrogenase 7 activity is essential for fetal de novo cholesterol synthesis and for neuroectodermal survival and cardiovascular differentiation in early mouse embryos. Endocrinology, 2010, 151: 1884-1892

[19] Larsen S, Holven K, Ottestad I, Dagsland K, Myhrstad M, Ulven S. Plasma fatty acid levels and gene expression related to lipid metabolism in peripheral blood mononuclear cells: a cross-sectional study in healthy subjects. Genes Nutr, 2018, 13: 9

[20] Leaver J, Villeneuve N, Obach A, Jensen L, Bron E, Tocher R, Taggart B. Functional genomics reveals increases in cholesterol biosynthetic genes and highly unsaturated fatty acid biosynthesis after dietary substitution of fish oil with vegetable oils in Atlantic salmon (Salmo salar). BMC Genomics, 2008, 9 299

[21] Leeson S, Caston L. Enrichment of eggs with lutein. Poult Sci, 2004, 83: 1709-1712

[22] Li J, Huang J, Li J, Chen H, Huang K, Zheng L. Accumulation of endoplasmic reticulum stress and lipogenesis in the liver through generational effects of high fat diets. J Hepatol, 2012, 56: 900-907

[23] Li K, Zhang J, Lyu H, Yang J, Wei W, Wang Y, Luo H, Zhang Y, Jiang X, Yi H, Wang M, Zhang C, Wu K, Xiao L, Wen W, Xu H, Li G, Wan Y, Yang F, Yang R, et al.. CSN6-SPOP-HMGCS1 axis promotes hepatocellular carcinoma progression via YAP1 activation. Adv Sci, 2024, 11: 2306827

[24] Lian C, Zhai Z, Li Z, Wang L. High fat diet-triggered non-alcoholic fatty liver disease: a review of proposed mechanisms. Chem Biol Interact, 2020, 330 109199

[25] Livak K, Schmittgen T. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 2001, 25: 402-408

[26] Long T, Debler E, Li X. Structural enzymology of cholesterol biosynthesis and storage. Curr Opin Struct Biol, 2022, 74 102369

[27] Luo J, Yang H, Song B. Mechanisms and regulation of cholesterol homeostasis. Nat Rev Mol Cell Biol, 2020, 21: 225-245

[28] Ma X, Bai Y, Liu K, Han Y, Zhang J, Liu Y, Hou X, Hao E, Hou Y, Bai G. Ursolic acid inhibits the cholesterol biosynthesis and alleviates high fat diet-induced hypercholesterolemia via irreversible inhibition of HMGCS1 in vivo. Phytomedicine, 2022, 103 154233

[29] Mansour A, El-feky M, El-Beltagi H, Sallam A. Synergism of dietary co-supplementation with lutein and bile salts improved the growth performance, carotenoid content, antioxidant capacity, lipid metabolism, and lipase activity of the marbled spinefoot rabbitfish, Siganus rivulatus. Animals, 2020, 10 1643

[30] Mattes R. Fat taste and lipid metabolism in humans. Physiol Behav, 2005, 86: 691-697

[31] Murillo A, Aguilar D, Norris G, DiMarco D, Missimer A, Hu S, Smyth J, Gannon S, Blesso C, Luo Y, Fernandez M. Compared with powdered lutein, a lutein nanoemulsion increases plasma and liver lutein, protects against hepatic steatosis, and affects lipoprotein metabolism in guinea pigs. J Nutr, 2016, 146: 1961-1969

[32] Naylor R, Hardy R, Buschmann A, Bush S, Cao L, Klinger D, Little D, Lubchenco J, Shumway S, Troell M. A 20-year retrospective review of global aquaculture. Nature, 2021, 591: 551-563

[33] Nelson J, Schmidt M, Askew E, Bernstein P, Von Tress M. Dietary modification and moderate antioxidant supplementation differentially affect serum carotenoids, antioxidant levels and markers of oxidative stress in older humans. J Nutr, 2003, 133: 3117-3123

[34] Nguyen P, Leray V, Diez M, Serisier S, Bloc’h J, Siliart B, Dumon H. Liver lipid metabolism. J Anim Physiol Anim Nutr, 2008, 92: 272-283

[35] Pan H, Chen H, Chen L, Zhang W, Lan P, Jia P, Li S, Yi M, Jia K. Establishment and characterization of a liver cell line from black porgy. Acanthopagrus Schlegelii Aquac Rep, 2022, 25101213

[36] Qiu X. Ameliorative effects of lutein on non-alcoholic fatty liver disease in rats. World J Gastroenterol, 2015, 21: 8061

[37] Quintana AM, Hernandez JA, Gonzalez CG. Functional analysis of the zebrafish ortholog of HMGCS1 reveals independent functions for cholesterol and isoprenoids in craniofacial development. PLoS ONE, 2017, 12 e0180856

[38] Renzi L, Hammond B, Dengler M, Roberts R. The relation between serum lipids and lutein and zeaxanthin in the serum and retina: results from cross-sectional, case-control and case study designs. Lipids Health Dis, 2012, 11: 33

[39] Ribaya-Mercado J, Blumberg J. Lutein and zeaxanthin and their potential roles in disease prevention. J Am Coll Nutr, 2004, 23: 567S-587S

[40] Schroor M, Mokhtar F, Plat J, Mensink R. Associations between SNPs in intestinal cholesterol absorption and endogenous cholesterol synthesis genes with cholesterol metabolism. Biomedicines, 2021, 9: 1475

[41] Shen Y, Li X, Bao Y, Zhu T, Wu Z, Yang B, Jiao L, Zhou Q, Jin M. Differential regulatory effects of optimal or excessive dietary lipid levels on growth, lipid metabolism and physiological response in black seabream (Acanthopagrus schlegelii). Aquaculture, 2022, 560 738532

[42] Shen Y, Zhao W, Monroig Ó, Bao Y, Zhu T, Jiao L, Sun P, Tocher D, Jin M. High-fat-diet-induced inflammation and apoptosis via activation of Ire1α in liver and hepatocytes of black seabream (Acanthopagrus schlegelii). Fish Shellfish Immunol, 2023, 143 109212

[43] Song F, Qin Y, Geng H, He C, Yang P, Wang W, Chen Y. Transcriptome analysis reveals the effects of dietary lipid level on growth performance and immune response in golden pompano (Trachinotus ovatus). Aquaculture, 2023, 563 738959

[44] Soundarya Priyadharsini K, Nirmala P, Ashok Kumar P, Krishna Prasad T. Effect of lutein in the expression of PPARα and LDLR in hypercholesterolemic male Wistar rats. Int J Basic Clin Pharmacol, 2018, 7: 684

[45] Teratani T, Tomita K, Suzuki T, Oshikawa T, Yokoyama H, Shimamura K, Tominaga S, Hiroi S, Irie R, Miura S, Hibi T. A high-cholesterol diet exacerbates liver fibrosis in mice via accumulation of free cholesterol in hepatic stellate cells. Gastroenterology, 2012, 142: 152-164.e10

[46] Tomita K, Teratani T, Suzuki T, Shimizu M, Sato H, Narimatsu K, Okada Y, Kurihara C, Irie R, Hibi T. Free cholesterol accumulation in hepatic stellate cells: mechanism of liver fibrosis aggravation in nonalcoholic steatohepatitis in mice. Hepatology, 2014, 59: 154-169

[47] Wang J, Wen Y, Zhao W, Zhang Y, Lin F, Ouyang C, Wang H, Yao L, Ma H, Zhao Y, Huang H, Shi X, Feng L, Lin D, Jiang B, Li Q. Hepatic conversion of acetyl-CoA to acetate plays crucial roles in energy stress. eLife, 2023, 12 RP87419

[48] Xiao M, Li F, Xie P, Qi Y, Xie J, Pei X. Gypenosides suppress hepatocellular carcinoma cells by blocking cholesterol biosynthesis through inhibition of MVA pathway enzyme HMGCS1. Chem Biol Interact, 2023, 383 110674

[49] Xu Z, Fan J, Ding X, Qiao L, Wang G. Characterization of high-fat, diet-induced, non-alcoholic steatohepatitis with fibrosis in rats. Dig Dis Sci, 2010, 55: 931-940

[50] Yan R, Ji S, Ku T, Sang N. Cross-omics analyses reveal the effects of ambient PM2.5 exposure on hepatic metabolism in female mice. Toxics, 2024, 12: 587

[51] Yang C, Zeisberg M, Mosterman B, Sudhakar A, Yerramalla U, Holthaus K, Xu L, Eng F, Afdhal N, Kalluri R. Liver fibrosis: insights into migration of hepatic stellate cells in response to extracellular matrix and growth factors. Gastroenterology, 2003, 124: 147-159

[52] Yi X, Li J, Xu W, Zhang W, Mai K. Effects of dietary lutein/canthaxanthin ratio on the growth and pigmentation of large yellow croaker Larimichthys croceus. Aquac Nutr, 2016, 22: 683-690

[53] Yousefi M, Adineh H, Ghafarifarsani H, Raeeszadeh M, Farsani M, Hashemianfar S. Immunological, antioxidant, growth responses, and disease resistance of rainbow trout, Oncorhynchus mykiss, with feeding diets supplemented with Lactobacillus salivarius and lutein. Ann Anim Sci, 2024, 24: 1211-1222

[54] Zheng Q, Li X, Huang N, Li F, Ge J, Wang D, Sun R, Liu R. Saikosaponins ameliorate hyperlipidemia in rats by enhancing hepatic lipid and cholesterol metabolism. J Ethnopharmacol, 2023, 305 116110

[55] Zhu T, Corraze G, Plagnes-Juan E, Skiba-Cassy S. Cholesterol metabolism regulation mediated by SREBP-2, LXRα and miR-33a in rainbow trout (Oncorhynchus mykiss) both in vivo and in vitro. PLoS ONE, 2020, 15 e0223813

[56] Zhu A, Xu D, Li Q, Li W, Zhang X, Yan X. Effects of ursodeoxycholic acid on growth performance and lipid metabolism in juvenile large yellow croaker (Larimichthys crocea). Aquaculture, 2024, 592 741166

Rights & permissions

The Author(s)

Just Accepted

This article has successfully passed peer review and final editorial review, and will soon enter typesetting, proofreading and other publishing processes. The currently displayed version is the accepted final manuscript. The officially published version will be updated with format, DOI and citation information upon launch. We recommend that you pay attention to subsequent journal notifications and preferentially cite the officially published version. Thank you for your support and cooperation.