Adipose-derived stem cell extracellular vesicles attenuate liver fibrosis via restoration of gut barrier function and modulation of gut microbiota

Abstract

Aim: Liver fibrosis (LF) is a major pathological stage that may progress to end-stage chronic liver injury but currently lacks effective treatment strategies. Previous studies have shown that adipose-derived stem cell extracellular vesicles (ADSC-EVs) play crucial roles in tissue repair, immune regulation, and anti-inflammatory effects. This study aims to elucidate the therapeutic effect of ADSC-EVs in LF and reveal their regulation mechanisms in gut-liver axis dysregulation.

Methods: The LF mouse model was established by intraperitoneal injection of diethylnitrosamine/CCl₄. LF mice for ADSC-EV treatment received ADSC-EVs (200 μg per mouse) twice a week for three weeks. Then, hepatic function tests, liver and gut histopathology, and gut microbiota analyses were performed.

Results: ADSC-EVs effectively improved hepatic function, reduced collagen deposition and suppressed hepatic stellate cell activation, exhibiting potent anti-fibrotic potential in LF mice. Additionally, they significantly restored intestinal barrier integrity by reducing intestinal permeability and reinforcing the mucus barrier. Furthermore, ADSC-EV treatment regulated gut microbiota dysbiosis, increased the abundance of beneficial intestinal bacteria such as Akkermansia muciniphila. ADSC-EV intervention also elevated the level of butyric acid in cecal contents and significantly reduced systemic inflammation.

Conclusion: Our findings suggest that ADSC-EVs represent a promising novel therapeutic strategy for LF, promoting liver tissue repair, enhancing intestinal barrier function, and maintaining gut homeostasis to establish a virtuous circle within the liver-gut axis.

Keywords

Liver fibrosis / ADSC-derived extracellular vesicles / gut microbiota / intestinal barrier / butyric acid / inflammation

References

[1] Karsdal MA,Daniels SJ,Krag A.Is the total amount as important as localization and type of collagen in liver fibrosis attributable to steatohepatitis?.Hepatology2020;71:346-51

[2] Villesen IF,Leeming DJ,Nielsen MJ.Review article: the signalling and functional role of the extracellular matrix in the development of liver fibrosis.Aliment Pharmacol Ther2020;52:85-97

[3] Zamani M,Ajmera V,Murad MH.Global prevalence of advanced liver fibrosis and cirrhosis in the general population: a systematic review and meta-analysis.Clin Gastroenterol Hepatol2025;23:1123-34

[4] Ferdek PE,Stopa KB.When healing turns into killing - the pathophysiology of pancreatic and hepatic fibrosis.J Physiol2022;600:2579-612

[5] Chen S,Wu X.Comparison of fibrosis regression of entecavir alone or combined with pegylated interferon alpha2a in patients with chronic hepatitis B.Hepatol Int2021;15:611-20

[6] Cai X,Xie W.Hydronidone for the treatment of liver fibrosis related to chronic hepatitis B: a phase 2 randomized controlled trial.Clin Gastroenterol Hepatol2023;21:1893-901.e7

[7] Rotman Y.Current and upcoming pharmacotherapy for non-alcoholic fatty liver disease.Gut2017;66:180-90

[8] Di Zeo-Sánchez DE,Pinazo-Bandera JM.Intestinal permeability and immune-inflammatory markers in patients with idiosyncratic drug-induced liver injury, drug-induced steatosis and metabolic dysfunction-associated steatotic liver disease (MASLD).Br J Pharmacol2025;182:5303-16

[9] Chopyk DM.Contribution of the intestinal microbiome and gut barrier to hepatic disorders.Gastroenterology2020;159:849-63 PMCID:PMC7502510

[10] Lau HC,Yu J.Gut microbiome in metabolic dysfunction-associated steatotic liver disease and associated hepatocellular carcinoma.Nat Rev Gastroenterol Hepatol2025;22:619-38

[11] He C,Luo Y.Exosome theranostics: biology and translational medicine.Theranostics2018;8:237-55 PMCID:PMC5743472

[12] Jeppesen DK,Franklin JL.Reassessment of exosome composition.Cell2019;177:428-45.e18 PMCID:PMC6664447

[13] Zhou Y,Xu W.Exosomes released by human umbilical cord mesenchymal stem cells protect against cisplatin-induced renal oxidative stress and apoptosis in vivo and in vitro.Stem Cell Res Ther2013;4:34 PMCID:PMC3707035

[14] Gatti S,Deregibus MC.Microvesicles derived from human adult mesenchymal stem cells protect against ischaemia-reperfusion-induced acute and chronic kidney injury.Nephrol Dial Transplant2011;26:1474-83

[15] Bruno S,Collino F.Microvesicles derived from mesenchymal stem cells enhance survival in a lethal model of acute kidney injury.PLoS One2012;7:e33115 PMCID:PMC3303802

[16] Zhang B,Gong A.HucMSC-exosome mediated-Wnt4 signaling is required for cutaneous wound healing.Stem Cells2015;33:2158-68

[17] Li T,Wang B.Exosomes derived from human umbilical cord mesenchymal stem cells alleviate liver fibrosis.Stem Cells Dev2013;22:845-54 PMCID:PMC3585469

[18] Jin Y,Li H.Extracellular vesicles secreted by human adipose-derived stem cells (hASCs) improve survival rate of rats with acute liver failure by releasing lncRNA H19.EBioMedicine2018;34:231-42 PMCID:PMC6116414

[19] Zhao S,Wang J.Engineering exosomes derived from subcutaneous fat MSCs specially promote cartilage repair as miR-199a-3p delivery vehicles in Osteoarthritis.J Nanobiotechnology2023;21:341 PMCID:PMC10515007

[20] Daemen S,Schilling JD.Comprehensive analysis of liver macrophage composition by flow cytometry and immunofluorescence in murine NASH.STAR Protoc2021;2:100511 PMCID:PMC8102804

[21] Wu B,Guo J.ADSCs-derived exosomes ameliorate hepatic fibrosis by suppressing stellate cell activation and remodeling hepatocellular glutamine synthetase-mediated glutamine and ammonia homeostasis.Stem Cell Res Ther2022;13:494 PMCID:PMC9531400

[22] Li C,Pu M,Lu Q.Coptisine alleviates colitis through modulating gut microbiota and inhibiting TXNIP/NLRP3 inflammasome.J Ethnopharmacol2024;335:118680

[23] Liu Y,Sun Z.Synergistic toxicity of combined exposure to acrylamide and polystyrene nanoplastics on the gut-liver axis in mice.Biology2025;14:523 PMCID:PMC12109039

[24] Aimaier D,Zhang Y.Susceptibility factor TNF-α synergizes with Polygonum multiflorum to drive idiosyncratic liver injury in mice by disrupting gut microbiota composition and hepatic metabolite homeostasis.J Inflamm Res2025;18:9477-94 PMCID:PMC12278978

[25] Ahmadi Badi S,Moradi HR.Comparative study of liver injury protection by Akkermansia muciniphila and Faecalibacterium prausnitzii interventions in live and cell-free supernatant forms via targeting the hepcidin - ferroportin axis in mice with CCl₄-induced liver fibrosis.Gut Pathog2025;17:54 PMCID:PMC12273000

[26] Oguri N,Nishinarita Y.Akkermansia muciniphila in the small intestine improves liver fibrosis in a murine liver cirrhosis model.NPJ Biofilms Microbiomes2024;10:81 PMCID:PMC11405509

[27] Ding Y,Lao X.The role of Akkermansia muciniphila in disease regulation.Probiotics Antimicrob Proteins2025;17:2027-38

[28] Mandaliya DK,Seshadri S.Postbiotic potential of SCFAs on metaflammation and gut microbiota alteration in diabetes.J Biosci2025;50:57

[29] Aydın MM.Liver fibrosis.Turk J Gastroenterol2018;29:14-21 PMCID:PMC6322608

[30] Hernandez-Gea V.Pathogenesis of liver fibrosis.Annu Rev Pathol2011;6:425-56

[31] Burki T.WHO’s 2024 global hepatitis report.Lancet Infect Dis2024;24:e362-3

[32] Gan C,Xue T.Focal adhesion kinase inhibitors in fibrotic diseases therapy: development and therapeutic potential.Eur J Med Chem2025;296:117882

[33] Kaya E,Oruc N.Non-invasive tests for resmetirom treatment fail to accurately define the target population: evidence from a biopsy-proven MASLD cohort.Hepatol Forum2025;6:111-5 PMCID:PMC12268769

[34] Adali G.Non-invasive tests fail to ensure therapeutic precision for resmetirom in MASLD.Hepatol Forum2025;6:89-90 PMCID:PMC12268762

[35] Wang W,Chen Y.TGF-β inhibitors: the future for prevention and treatment of liver fibrosis?.Front Immunol2025;16:1583616 PMCID:PMC12245701

[36] Dumitru A,Bădescu AC.Linking gut permeability to liver steatosis: noninvasive biomarker evaluation in MASLD patients - a prospective cross-sectional study.Medicine2025;104:e42476 PMCID:PMC12114047

[37] Cui C,Shi J.Gut-liver axis: the role of intestinal microbiota and their metabolites in the progression of metabolic dysfunction-associated steatotic liver disease.Gut Liver2025;19:479-507 PMCID:PMC12261135

[38] Jasirwan COM,Hasan I,Rinaldi I.Correlation of gut Firmicutes/Bacteroidetes ratio with fibrosis and steatosis stratified by body mass index in patients with non-alcoholic fatty liver disease.Biosci Microbiota Food Health2021;40:50-8 PMCID:PMC7817510

[39] Mantovani A,Thoen RU.Firmicutes/Bacteroidetes and Firmicutes/Proteobacteria ratios are associated with worse prognosis in a cohort of Latin American patients with cirrhosis.Clinics2024;79:100471 PMCID:PMC11345307

[40] Masihi KN,Lange W,Ribi E.Trehalose dimycolate from various mycobacterial species induces differing anti-infectious activities in combination with muramyl dipeptide.Infect Immun1985;50:938-40 PMCID:PMC261175

[41] Piao C,Kou Z.Effects of exosomes derived from adipose-derived mesenchymal stem cells on pyroptosis and regeneration of injured liver.Int J Mol Sci2022;23:12065 PMCID:PMC9602906

[42] Choi H,Kim DH.Quantitative biodistribution and pharmacokinetics study of GMP-grade exosomes labeled with ⁸⁹Zr radioisotope in mice and rats.Pharmaceutics2022;14:1118 PMCID:PMC9229812

[43] Yu JX,Chen X.Gut microbiota in liver diseases: initiation, development and therapy.Front Med2025;12:1615839 PMCID:PMC12174154

[44] Zhang YL,Gou HZ,Zhang L.The gut microbiota-bile acid axis: a potential therapeutic target for liver fibrosis.Front Cell Infect Microbiol2022;12:945368 PMCID:PMC9519863

[45] Pisarello MJL,Chaia AP.Targeting gut health: probiotics as promising therapeutics in alcohol-related liver disease management.AIMS Microbiol2025;11:410-35 PMCID:PMC12207258

[46] Cai Z,Li J.Treatment of inflammatory bowel disease: a comprehensive review.Front Med2021;8:765474 PMCID:PMC8720971