{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T06:09:53Z","timestamp":1790143793692,"version":"4.0.1"},"reference-count":30,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T00:00:00Z","timestamp":1784592000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Gut barrier dysfunction and portal endotoxemia contribute to metabolic dysfunction-associated steatohepatitis (MASH) progression through activation of hepatic inflammatory signaling. Milk-derived extracellular vesicles (EVs) contain bioactive microRNAs and have recently attracted attention as modulators of intestinal homeostasis. This study investigated the effect of bovine milk-derived extracellular vesicles (B-mEVs) in ameliorating MASH by improving intestinal barrier function. C57BL\/6J mice fed a choline-deficient amino acid-defined high-fat diet (CDA-HFD) were orally treated with B-mEVs, and therapeutic effects were evaluated. Liver histology, fibrosis, portal lipopolysaccharide (LPS) levels, intestinal permeability, and gut microbiota composition were evaluated. The direct effects of B-mEVs on intestinal barrier function were assessed using palmitic acid-stimulated Caco-2 cells. Small RNA sequencing and microRNA enrichment analyses were performed to characterize B-mEV cargo. B-mEV treatment attenuated hepatic steatosis, inflammation, and fibrosis in CDA-HFD-fed mice and reduced serum aminotransferase levels, portal LPS concentrations, hepatic macrophage accumulation, and hepatic TLR4\/NF-\u03baB signaling activation. Meanwhile, B-mEVs restored intestinal tight junction proteins, including ZO-1, occludin, and claudin-1, and improved intestinal permeability in vivo. In Caco-2 cells, B-mEVs attenuated palmitic acid-induced barrier dysfunction and suppressed myosin light chain kinase expression. Gut microbiota analysis revealed partial restoration of Akkermansia abundance after B-mEV administration. Furthermore, to explore the molecular basis of these protective effects, small RNA sequencing demonstrated enrichment of regulatory microRNAs, including let-7a-5p, and pathway analyses identified associations with intestinal barrier and inflammatory signaling pathways. B-mEVs ameliorated CDA-HFD-induced steatohepatitis by restoring intestinal barrier integrity and suppressing gut-derived LPS\/TLR4 inflammatory signaling. These findings suggested that milk EVs may represent a novel gut\u2013liver axis-targeted therapeutic strategy for MASH.<\/jats:p>","DOI":"10.3390\/ijms27146485","type":"journal-article","created":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T15:17:52Z","timestamp":1784647072000},"page":"6485","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Bovine Milk-Derived Extracellular Vesicles Ameliorate Steatohepatitis by Restoring Gut Barrier in CDA-HFD-Fed Mice"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-5703-016X","authenticated-orcid":false,"given":"Tatsuya","family":"Nakatani","sequence":"first","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3049-3443","authenticated-orcid":false,"given":"Shinya","family":"Sato","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kosuke","family":"Kaji","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroki","family":"Kachi","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naoki","family":"Nishimura","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masafumi","family":"Oyama","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-9174-2477","authenticated-orcid":false,"given":"Jun-ichi","family":"Hanatani","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3575-5609","authenticated-orcid":false,"given":"Satoshi","family":"Iwai","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soichi","family":"Takeda","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6295-3283","authenticated-orcid":false,"given":"Norihisa","family":"Nishimura","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5794-1512","authenticated-orcid":false,"given":"Koh","family":"Kitagawa","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3158-5318","authenticated-orcid":false,"given":"Tadashi","family":"Namisaki","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hitoshi","family":"Yoshiji","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Nara Medical University, 840 Shijo-cho, Kashihara 634-8522, Nara, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1547","DOI":"10.1002\/hep.27368","article-title":"Fibrosis stage is the strongest predictor for disease-specific mortality in NAFLD after up to 33 years of follow-up","volume":"61","author":"Ekstedt","year":"2015","journal-title":"Hepatology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1265","DOI":"10.1016\/j.jhep.2017.07.027","article-title":"Fibrosis stage but not NASH predicts mortality and time to development of severe liver disease in biopsy-proven NAFLD","volume":"67","author":"Hagstrom","year":"2017","journal-title":"J. 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