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Microbiota dysbiosis occurs when energy imbalances appear due to an unhealthy diet and a sedentary lifestyle. Dysbiosis has a critical impact on increasing intestinal permeability and epithelial barrier deterioration, contributing to bacterial and antigen translocation to the liver, triggering non-alcoholic fatty liver disease (NAFLD) progression. In this study, the potential therapeutic\/beneficial effects of a combination of metabolic cofactors (a multi-ingredient; MI) (betaine, N-acetylcysteine, L-carnitine, and nicotinamide riboside) against NAFLD were evaluated. In addition, we investigated the effects of this metabolic cofactors\u2019 combination as a modulator of other players of the gut-liver axis during the disease, including gut barrier dysfunction and microbiota dysbiosis. Diet-induced NAFLD mice were distributed into two groups, treated with the vehicle (NAFLD group) or with a combination of metabolic cofactors (NAFLD-MI group), and small intestines were harvested from all animals for histological, molecular, and omics analysis. The MI treatment ameliorated gut morphological changes, decreased gut barrier permeability, and reduced gene expression of some proinflammatory cytokines. Moreover, epithelial cell proliferation and the number of goblet cells were increased after MI supplementation. In addition, supplementation with the MI combination promoted changes in the intestinal microbiota composition and diversity, as well as modulating short-chain fatty acids (SCFAs) concentrations in feces. Taken together, this specific combination of metabolic cofactors can reverse gut barrier disruption and microbiota dysbiosis contributing to the amelioration of NAFLD progression by modulating key players of the gut-liver axis.<\/jats:p>","DOI":"10.3390\/ijms232213675","type":"journal-article","created":{"date-parts":[[2022,11,8]],"date-time":"2022-11-08T08:52:56Z","timestamp":1667897576000},"page":"13675","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":50,"title":["Microbiota Dysbiosis and Gut Barrier Dysfunction Associated with Non-Alcoholic Fatty Liver Disease Are Modulated by a Specific Metabolic Cofactors\u2019 Combination"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6588-1193","authenticated-orcid":false,"given":"Sergio","family":"Quesada-V\u00e1zquez","sequence":"first","affiliation":[{"name":"Eurecat, Technology Centre of Catalunya, Nutrition and Health Unit, 43204 Reus, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caitlin","family":"Bone","sequence":"additional","affiliation":[{"name":"Gut Microbes and Health Institute Strategic Programme, Quadram Institute Bioscience, Norwich Research Park, Norwich NR4 7UQ, Norfolk, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shikha","family":"Saha","sequence":"additional","affiliation":[{"name":"Food Innovation and Health Institute Strategic Programme, Quadram Institute Bioscience, Norwich Research Park, Norwich NR4 7UQ, Norfolk, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iris","family":"Triguero","sequence":"additional","affiliation":[{"name":"Eurecat, Technology Centre of Catalunya, Nutrition and Health Unit, 43204 Reus, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1760-5175","authenticated-orcid":false,"given":"Marina","family":"Colom-Pellicer","sequence":"additional","affiliation":[{"name":"Nutrigenomics Research Group, Department of Biochemistry and Biotechnology, Universitat Rovira i Virgili, 43007 Tarragona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8657-5726","authenticated-orcid":false,"given":"Gerard","family":"Aragon\u00e8s","sequence":"additional","affiliation":[{"name":"Nutrigenomics Research Group, Department of Biochemistry and Biotechnology, Universitat Rovira i Virgili, 43007 Tarragona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0078-8948","authenticated-orcid":false,"given":"Falk","family":"Hildebrand","sequence":"additional","affiliation":[{"name":"Gut Microbes and Health Institute Strategic Programme, Quadram Institute Bioscience, Norwich Research Park, Norwich NR4 7UQ, Norfolk, UK"},{"name":"Digital Biology, Earlham Institute, Norwich Research Park, Norwich NR4 7UZ, Norfolk, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0700-2004","authenticated-orcid":false,"given":"Josep M.","family":"del Bas","sequence":"additional","affiliation":[{"name":"Eurecat, Centre Tecnol\u00f2gic de Catalunya, Biotechnology Area, 43204 Reus, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6144-0294","authenticated-orcid":false,"given":"Antoni","family":"Caimari","sequence":"additional","affiliation":[{"name":"Eurecat, Centre Tecnol\u00f2gic de Catalunya, Biotechnology Area, 43204 Reus, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naiara","family":"Beraza","sequence":"additional","affiliation":[{"name":"Gut Microbes and Health Institute Strategic Programme, Quadram Institute Bioscience, Norwich Research Park, Norwich NR4 7UQ, Norfolk, UK"},{"name":"Food Innovation and Health Institute Strategic Programme, Quadram Institute Bioscience, Norwich Research Park, Norwich NR4 7UQ, Norfolk, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1172-3995","authenticated-orcid":false,"given":"Xavier","family":"Escot\u00e9","sequence":"additional","affiliation":[{"name":"Eurecat, Technology Centre of Catalunya, Nutrition and Health Unit, 43204 Reus, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ji, Y., Yin, Y., Sun, L., and Zhang, W. 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