Mechanism of arbutin in metabolic dysfunction-associated fatty liver disease based on multi-omics research

Abstract

Introduction

Gut microbiota regulation is a key strategy for treating metabolic dysfunction-associated fatty liver disease (MAFLD). Arbutin (ARB) is a natural hydroquinone active agent with anti-inflammatory and antioxidant effects, as well as regulatory effects on the gut microbiota. However, its therapeutic effect on MAFLD and the responsible mechanisms remain unclear.

Objectives

This study explored the therapeutic effect and mechanisms of ARB in MAFLD treatment.

Methods

High-fat diet (HFD)-fed mice served as the in vivo MAFLD model, and ARB treatment was given simultaneously. The extent of liver injury was assessed through histopathological staining. AML12 cells treated with free fatty acids served as the in vitro model. The effects of ARB were evaluated via oil red O staining and biochemical assays. Subsequently, we utilized bioinformatics techniques to predict the potential mechanisms and targets of ARB. The expression of liver apoptosis-related genes was detected using molecular biology techniques. Alterations in the gut microbiota were analyzed by 16S rRNA sequencing. Ultrahigh-performance liquid chromatography–high-resolution mass spectrometry was used to analyze the changes in fecal metabolite levels.

Results

ARB treatment effectively improved liver injury in mice with MAFLD. Its mechanism was associated with anti-apoptotic effects mediated by signal transducer and activator of transcription 3. Meanwhile, ARB effectively reversed gut microbiota imbalance in mice with MAFLD and altered the composition of gut microbes and fecal metabolites.

Conclusion

ARB displayed potential effects in alleviating the pathology of MAFLD, exerting anti-apoptotic actions, and restoring the gut microbiota balance.

Graphical Abstract

Keywords

MAFLD / Arbutin / Apoptosis / Gut microbiota / Fecal metabolites

References

[1] Bedi O, Aggarwal S, Trehanpati N, Ramakrishna G, Grewal AS, Krishan P (2020) In vitro targeted screening and molecular docking of stilbene, quinones, and flavonoid on 3T3-L1 pre-adipocytes for anti-adipogenic actions. Naunyn-Schmiedeberg’s archives of pharmacology 393. 2093–2106. https://doi.org/10.1007/s00210-020-01919-w

[2] Brunt EM, Kleiner DE, Wilson LA, Belt P, Neuschwander-Tetri BA (2011) Nonalcoholic fatty liver disease (NAFLD) activity score and the histopathologic diagnosis in NAFLD: distinct clinicopathologic meanings. Hepatol (Baltimore Md) 53 810–820. https://doi.org/10.1002/hep.24127

[3] Byrne CD, Targher G. NAFLD: a multisystem disease. J Hepatol. 2015, 62: 47-64.

[4] Daina A, Michielin O, Zoete V (2019) SwissTargetPrediction: updated data and new features for efficient prediction of protein targets of small molecules. 47:W357–w364. Nucleic acids research10.1093/nar/gkz382

[5] den Besten G, Lange K, Havinga R, van Dijk TH, Gerding A, van Eunen K, Müller M, Groen AK, Hooiveld GJ, Bakker BM et al (2013) Gut-derived short-chain fatty acids are vividly assimilated into host carbohydrates and lipids. American journal of physiology. Gastrointestinal and liver physiology 305. G900–910. https://doi.org/10.1152/ajpgi.00265.2013

[6] Dennis G Jr., Sherman BT, Hosack DA, Yang J, Gao W, Lane HC, Lempicki RA (2003) DAVID: Database for Annotation, Visualization, and Integrated Discovery. 4:P3Genome biology

[7] Di Vincenzo F, Del Gaudio A, Petito V, Lopetuso LR, Scaldaferri F. Gut microbiota, intestinal permeability, and systemic inflammation: a narrative review. Intern Emerg Med. 2024, 19: 275-293.

[8] Dong X, Lu S, Tian Y, Ma H, Wang Y, Zhang X, Sun G, Luo Y, Sun X. Bavachinin protects the liver in NAFLD by promoting regeneration via targeting PCNA. J Adv Res. 2024, 55: 131-144.

[9] Gallo K, Goede A, Preissner R, Gohlke BO (2022) SuperPred 3.0: drug classification and target prediction-a machine learning approach. 50:W726–w731. Nucleic acids research10.1093/nar/gkac297

[10] Jiang T, Xiao Y, Zhou J, Luo Z, Yu L, Liao Q, Liu S, Qi X, Zhang H, Hou Met al. . Arbutin alleviates fatty liver by inhibiting ferroptosis via FTO/SLC7A11 pathway. Redox Biol. 2023, 68: 102963.

[11] Jiang Y, Luo P, Cao Y, Peng D, Huo S, Guo J, Wang M, Shi W, Zhang C, Li Set al. . The role of STAT3/VAV3 in glucolipid metabolism during the development of HFD-induced MAFLD. Int J Biol Sci. 2024, 202027-2043.

[12] Jiang J, Gao Y, Wang J, Huang Y, Yang R, Zhang Y, Ma Y, Wen Y, Luo G, Zhang Set al. . Hepatic sphingomyelin phosphodiesterase 3 promotes steatohepatitis by disrupting membrane sphingolipid metabolism. Cell Metabol. 2025, 37: 1119-1136e1113.

[13] Kilkenny C, Browne WJ, Cuthill IC, Emerson M, Altman DG. Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. PLoS Biol. 2010, 8: e1000412.

[14] Li X, Wang J, Gong X, Zhang M, Kang S, Shu B, Wei Z, Huang ZS, Li D (2020) Upregulation of BCL-2 by acridone derivative through gene promoter i-motif for alleviating liver damage of NAFLD/NASH. Nucleic acids research. 48:8255–8268. https://doi.org/10.1093/nar/gkaa615

[15] Liu Y, Yang X, Gan J, Chen S, Xiao ZX, Cao Y (2022) CB-Dock2: improved protein-ligand blind docking by integrating cavity detection, docking and homologous template fitting. Nucleic acids research 50. W159–w164. https://doi.org/10.1093/nar/gkac394

[16] Liu J, Wang D, Xie Z, Ding L, Li S, Ma X, Liu J, Ren J, Xiao C, Yang Cet al. . Combination of Pioglitazone and Metformin Actions on Liver Lipid Metabolism in Obese Mice. Biomolecules 13. 2023.

[17] Liu T, Cheng Z, Song D, Zhu E, Li H, Lin R, Wan Z, Liu S, Gong Z, Shan C (2024) Arbutin alleviates Mycoplasma gallinarum-induced damage caused by pulmonary fibrosis via the JAK2/STAT3 pathway. 103:104434. Poultry science10.1016/j.psj.2024.104434

[18] Ma J, Chen S, Li Y, Wu X, Song Z. Arbutin improves gut development and serum lipids via Lactobacillus intestinalis. Front Nutr. 2022, 9948573.

[19] Madić V, Petrović A, Jušković M, Jugović D, Djordjević L, Stojanović G, Vasiljević P. Polyherbal mixture ameliorates hyperglycemia, hyperlipidemia and histopathological changes of pancreas, kidney and liver in a rat model of type 1 diabetes. J Ethnopharmacol. 2021, 265113210.

[20] Mao L, Lu J, Yang Q, Liu Z, Wu C, Ke B, Su K, Yuan H, Cui Y, Wang Y et al (2025) Bilirubin Targeting WNK1 to Alleviate NLRP3-Mediated Neuroinflammation. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 12, e2407349. https://doi.org/10.1002/advs.202407349

[21] Marra F, Svegliati-Baroni G. Lipotoxicity and the gut-liver axis in NASH pathogenesis. J Hepatol. 2018, 68280-295.

[22] Meng X, Li S, Li Y, Gan RY, Li HB. Gut Microbiota’s Relationship with Liver Disease and Role in Hepatoprotection by Dietary Natural Products and Probiotics. Nutrients 10. 2018.

[23] Min HK, Mirshahi F, Verdianelli A, Pacana T, Patel V, Park CG, Choi A, Lee JH, Park CB, Ren S et al (2015) Activation of the GP130-STAT3 axis and its potential implications in nonalcoholic fatty liver disease. American journal of physiology. Gastrointestinal and liver physiology 308. G794–803. https://doi.org/10.1152/ajpgi.00390.2014

[24] Mota M, Banini BA, Cazanave SC, Sanyal AJ. Molecular mechanisms of lipotoxicity and glucotoxicity in nonalcoholic fatty liver disease. Metab Clin Exp. 2016, 65: 1049-1061.

[25] Pabst O, Hornef MW, Schaap FG, Cerovic V, Clavel T, Bruns T. Gut-liver axis: barriers and functional circuits. Nat Rev Gastroenterol Hepatol. 2023, 20: 447-461.

[26] Park J, Zhao Y, Zhang F, Zhang S, Kwong AC, Zhang Y, Hoffmann HH, Bushweller L, Wu X, Ashbrook AWet al. . IL-6/STAT3 axis dictates the PNPLA3-mediated susceptibility to non-alcoholic fatty liver disease. J Hepatol. 2023, 78: 45-56.

[27] Qi W, Cao X, Chen Y, Chen H, Zhang N, Liu R, Wang W, Liu Q, Zheng S, Li Set al. . JiGuCao capsule formula alleviates metabolic fatty liver disease by regulating the gut-liver axis and lipid metabolism. Phytomedicine: Int J phytotherapy phytopharmacology. 2025, 140: 156559.

[28] Rong L, Zou J, Ran W, Qi X, Chen Y, Cui H, Guo J. Advancements in the treatment of non-alcoholic fatty liver disease (NAFLD). Front Endocrinol. 2022, 131087260.

[29] Shu Y, He D, Li W, Wang M, Zhao S, Liu L, Cao Z, Liu R, Huang Y, Li Het al. . Hepatoprotective Effect of Citrus aurantium L. Against APAP-induced Liver Injury by Regulating Liver Lipid Metabolism and Apoptosis. Int J Biol Sci. 2020, 16752-765.

[30] Sun Y, Zhang S, Nie Q, He H, Tan H, Geng F, Ji H, Hu J, Nie S. Gut firmicutes: Relationship with dietary fiber and role in host homeostasis. Crit Rev Food Sci Nutr. 2023, 63: 12073-12088.

[31] Szklarczyk D, Kirsch R, Koutrouli M, Nastou K, Mehryary F, Hachilif R, Gable AL, Fang T, Doncheva NT, Pyysalo Set al. . The STRING database in 2023: protein-protein association networks and functional enrichment analyses for any sequenced genome of interest. Nucleic Acids Res. 2023, 51: D638-d646.

[32] Wang X, Pan C, Gong J, Liu X, Li H. Enhancing the Enrichment of Pharmacophore-Based Target Prediction for the Polypharmacological Profiles of Drugs. J Chem Inf Model. 2016, 56: 1175-1183.

[33] Yang JW, Zou Y, Chen J, Cui C, Song J, Yang MM, Gao J, Hu HQ, Xia LQ, Wang LMet al. . Didymin alleviates metabolic dysfunction-associated fatty liver disease (MAFLD) via the stimulation of Sirt1-mediated lipophagy and mitochondrial biogenesis. J translational Med. 2023, 21: 921.

[34] Zhou H, Zhao J, Li A, Reetz MT (2019) Chemical and Biocatalytic Routes to Arbutin (†). Molecules 24. https://doi.org/10.3390/molecules24183303

[35] Zhu J, Zhou M, Zhao X, Mu M, Cheng M. Blueberry, combined with probiotics, alleviates non-alcoholic fatty liver disease via IL-22-mediated JAK1/STAT3/BAX signaling. Food Funct. 2018, 96298-6306.

[36] Zhu Y, Chen B, Zhang X, Akbar MT, Wu T, Zhang Y, Zhi L, Shen Q. Exploration of the Muribaculaceae Family in the Gut Microbiota: Diversity, Metabolism, and Function. Nutrients 16. 2024.

[37] Zou HY, Zhang HJ, Zhao YC, Li XY, Wang YM, Zhang TT, Xue CH (2023) N-3 PUFA Deficiency Aggravates Streptozotocin-Induced Pancreatic Injury in Mice but Dietary Supplementation with DHA/EPA Protects the Pancreas via Suppressing Inflammation, Oxidative Stress and Apoptosis. Mar Drugs 21. https://doi.org/10.3390/md21010039

Funding

Special Fund for Science and Technology Innovation Teams of Shanxi Province(No. 202304051001021)

Fundamental research program of Shanxi province(No. 20210302123289)

the National Natural Science Foundation of China (No.82270915)

RIGHTS & PERMISSIONS

The Author(s)