{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T16:16:40Z","timestamp":1788970600865,"version":"build-2803163510"},"reference-count":53,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T00:00:00Z","timestamp":1788912000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Metabolic dysfunction-associated steatotic liver disease may be influenced by human gut microbiota-derived metabolites. Fructooligosaccharides (FOS) and probiotic\n                      <jats:italic>Lactobacillus<\/jats:italic>\n                      species have been reported to be associated with metabolic improvements. However, the strain-specific contribution of defined probiotic consortia and the interplay between substrate-driven microbial metabolites and hepatic lipid regulation remain insufficiently characterized. This study aims to investigate whether FOS-driven metabolism of a three-strain\n                      <jats:italic>Lactobacillus<\/jats:italic>\n                      consortium (\n                      <jats:italic>L. plantarum<\/jats:italic>\n                      PBS067,\n                      <jats:italic>L. acidophilus<\/jats:italic>\n                      PBS066, and\n                      <jats:italic>L. reuteri<\/jats:italic>\n                      PBS072) modulates hepatocyte lipid accumulation in steatotic HepG2 cells within a reconstructed reductionist\n                      <jats:italic>in vitro<\/jats:italic>\n                      human gut microbiota model.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      A previously validated minimal core community, representing metabolic functions of the human gut microbiota, was supplemented with the probiotic consortium and cultured in the presence of FOS with an average degree of polymerization ~10.\n                      <jats:italic>Lactobacillus<\/jats:italic>\n                      strain growth and modulation, transcriptional activation of FOS-related genes, and short-chain fatty acid (SCFA) metabolic profiles were characterized. Microbiota-derived metabolites were then tested on palmitate-induced steatotic HepG2 cells.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results and discussion<\/jats:title>\n                    <jats:p>\n                      FOS supported growth of both probiotic and minimal core members in a strain-dependent manner within the community, with higher growth levels of\n                      <jats:italic>L. plantarum<\/jats:italic>\n                      PBS067 and\n                      <jats:italic>L. reuteri<\/jats:italic>\n                      PBS072. RT-qPCR analyses of\n                      <jats:italic>sacA, scrB<\/jats:italic>\n                      , and\n                      <jats:italic>sacK1<\/jats:italic>\n                      genes revealed species-specific transcriptional activation and context-dependent regulation of FOS utilization pathways. Microbial metabolic profiling showed production of SCFAs and organic acids typical of carbohydrate fermentation. Treatment of steatotic HepG2 cells with these metabolites significantly reduced intracellular lipid accumulation and down-regulated Cd36 gene encoding a key fatty acid transporter involved in palmitic acid-induced steatosis. These findings suggest that FOS-driven microbial metabolism within a defined\n                      <jats:italic>Lactobacillus<\/jats:italic>\n                      consortium could be associated with hepatocyte lipid uptake under controlled\n                      <jats:italic>in vitro<\/jats:italic>\n                      conditions, highlighting the interplay among microbial transcriptional activation and their metabolic profiles.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1866842","type":"journal-article","created":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T15:47:48Z","timestamp":1788968868000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Fructooligosaccharides-driven metabolism of a defined Lactobacillus probiotic consortium modulates hepatic lipid accumulation in a reductionist in vitro gut microbiota\u2013liver interaction model"],"prefix":"10.3389","volume":"17","author":[{"given":"Margherita","family":"Finazzi","sequence":"first","affiliation":[{"name":"Department of Biotechnology and Biosciences, University of Milano-Bicocca","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federica","family":"Bovio","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biosciences, University of Milano-Bicocca","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matilde","family":"Forcella","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biosciences, University of Milano-Bicocca","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Lasagni","sequence":"additional","affiliation":[{"name":"Department of Earth and Environmental Sciences, University of Milano-Bicocca","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola","family":"Fusi","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biosciences, University of Milano-Bicocca","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Zampolli","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biosciences, University of Milano-Bicocca","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrizia","family":"Di Gennaro","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biosciences, University of Milano-Bicocca","place":["Milano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,9]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"3011","DOI":"10.3390\/ijerph16173011","article-title":"Diet and non-alcoholic fatty liver disease: the Mediterranean way","volume":"16","author":"Abenavoli","year":"2019","journal-title":"Int. J. Environ. Res. Public Health"},{"key":"B2","doi-asserted-by":"publisher","first-page":"403","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic local alignment search tool","volume":"215","author":"Altschul","year":"1990","journal-title":"J. Mol. Biol"},{"key":"B3","doi-asserted-by":"publisher","first-page":"e44409","DOI":"10.1371\/journal.pone.0044409","article-title":"Transcriptional analysis of prebiotic uptake and catabolism by Lactobacillus acidophilus NCFM","volume":"7","author":"Andersen","year":"2012","journal-title":"PLos ONE"},{"key":"B4","doi-asserted-by":"publisher","first-page":"8957","DOI":"10.1073\/pnas.1332765100","article-title":"Functional and comparative genomic analyses of an operon involved in fructooligosaccharide utilization by Lactobacillus acidophilus","volume":"100","author":"Barrangou","year":"2003","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B5","doi-asserted-by":"publisher","first-page":"3816","DOI":"10.1073\/pnas.0511287103","article-title":"Global analysis of carbohydrate utilization by Lactobacillus acidophilus using cDNA microarrays","volume":"103","author":"Barrangou","year":"2006","journal-title":"Proc. Natl. Acad. Sci."},{"key":"B6","doi-asserted-by":"publisher","first-page":"104816","DOI":"10.1016\/j.micinf.2021.104816","article-title":"Defined gut microbial communities: promising tools to understand and combat disease","volume":"23","author":"Bayer","year":"2021","journal-title":"Microbes Infect."},{"key":"B7","doi-asserted-by":"publisher","first-page":"715","DOI":"10.4254\/wjh.v9.i16.715","article-title":"Non-alcoholic fatty liver disease: an expanded review","volume":"9","author":"Benedict","year":"2017","journal-title":"World J. Hepatol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1038\/nrgastro.2015.47","article-title":"Towards a more comprehensive concept for prebiotics","volume":"12","author":"Bindels","year":"2015","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"261","DOI":"10.1038\/s41574-019-0156-z","article-title":"Gut microbial metabolites in obesity, NAFLD and T2DM","volume":"15","author":"Canfora","year":"2019","journal-title":"Nat. Rev. Endocrinol."},{"key":"B10","doi-asserted-by":"publisher","first-page":"2374","DOI":"10.1007\/s00125-007-0791-0","article-title":"Selective increases of bifidobacteria in gut microflora improve high-fat-diet-induced diabetes in mice through a mechanism associated with endotoxaemia","volume":"50","author":"Cani","year":"2007","journal-title":"Diabetologia"},{"key":"B11","doi-asserted-by":"publisher","first-page":"1071284","DOI":"10.3389\/fnut.2022.1071284","article-title":"Lactobacillus plantarum ZJUIDS14 alleviates non-alcoholic fatty liver disease in mice in association with modulation in the gut microbiota","volume":"9","author":"Cao","year":"2023","journal-title":"Front. Nutr."},{"key":"B12","doi-asserted-by":"publisher","first-page":"7697","DOI":"10.1128\/AEM.02426-15","article-title":"Metabolism of fructooligosaccharides in Lactobacillus plantarum ST-III via differential gene transcription and alteration of cell membrane fluidity","volume":"81","author":"Chen","year":"2015","journal-title":"Appl. Environ. Microbiol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"13452","DOI":"10.3390\/ijms222413452","article-title":"The carbohydrate metabolism of Lactiplantibacillus plantarum","volume":"22","author":"Cui","year":"2021","journal-title":"Int. J. Mol. Sci."},{"key":"B14","doi-asserted-by":"publisher","first-page":"373","DOI":"10.3390\/ijms24010373","article-title":"Synergistic antioxidant effect of prebiotic ginseng berries extract and probiotic strains on healthy and tumoral colorectal cell lines","volume":"24","author":"De Giani","year":"","journal-title":"Int. J. Mol. Sci."},{"key":"B15","doi-asserted-by":"publisher","first-page":"e0301822","DOI":"10.1371\/journal.pone.0301822","article-title":"Positive modulation of a new reconstructed human gut microbiota by Maitake extract helpfully boosts the intestinal environment in vitro","volume":"19","author":"De Giani","year":"2024","journal-title":"PLoS ONE"},{"key":"B16","doi-asserted-by":"publisher","first-page":"1256","DOI":"10.3390\/microorganisms10061256","article-title":"Effects of inulin-based prebiotics alone or in combination with probiotics on human gut microbiota and markers of immune system: a randomized, double-blind, placebo-controlled study in healthy subjects","volume":"10","author":"De Giani","year":"","journal-title":"Microorganisms"},{"key":"B17","doi-asserted-by":"publisher","first-page":"1472","DOI":"10.1111\/1462-2920.12343","article-title":"Synthetic microbial ecosystems: an exciting tool to understand and apply microbial communities","volume":"16","author":"De Roy","year":"2014","journal-title":"Environ. Microbiol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1128\/mBio.00095-20","article-title":"Synergy between cell surface glycosidases and glycan-binding proteins dictates the utilization of specific beta (1, 3)-glucans by human gut Bacteroides","volume":"11","author":"D\u00e9jean","year":"2020","journal-title":"MBio."},{"key":"B19","doi-asserted-by":"publisher","first-page":"639","DOI":"10.1038\/nrendo.2011.126","article-title":"Targeting gut microbiota in obesity: effects of prebiotics and probiotics","volume":"7","author":"Delzenne","year":"2011","journal-title":"Nat. Rev. Endocrinol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"202","DOI":"10.1016\/j.jhep.2020.03.039","article-title":"A new definition for metabolic dysfunction-associated fatty liver disease: an international expert consensus statement","volume":"73","author":"Eslam","year":"2020","journal-title":"J. Hepatol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"2974","DOI":"10.3748\/wjg.v24.i27.2974","article-title":"Pathogenesis of non-alcoholic fatty liver disease in children and adolescence: from \u201ctwo hit theory\u201d to \u201cmultiple hit model\u201d","volume":"24","author":"Fang","year":"2018","journal-title":"World J. Gastroenterol."},{"key":"B22","doi-asserted-by":"publisher","first-page":"192","DOI":"10.3390\/microorganisms13010192","article-title":"Beneficial effects of in vitro reconstructed human gut microbiota by ginseng extract fermentation on intestinal cell lines","volume":"13","author":"Finazzi","year":"2025","journal-title":"Microorganisms"},{"key":"B23","doi-asserted-by":"publisher","first-page":"e1001314","DOI":"10.1371\/journal.pgen.1001314","article-title":"The evolution of host specialization in the vertebrate gut symbiont Lactobacillus reuteri","volume":"7","author":"Frese","year":"2011","journal-title":"PLoS Genet."},{"key":"B24","doi-asserted-by":"publisher","first-page":"96","DOI":"10.1016\/S0016-5085(70)80098-1","article-title":"Carbohydrate digestion and absorption","volume":"58","author":"Gray","year":"1970","journal-title":"Gastroenterology"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.29219\/fnr.v64.3751","article-title":"Bile salt hydrolase-overexpressing Lactobacillus strains can improve hepatic lipid accumulation in vitro in an NAFLD cell model","author":"Huang","year":"2020","journal-title":"Food Nutr. Res"},{"key":"B26","doi-asserted-by":"publisher","first-page":"114300","DOI":"10.1016\/j.biopha.2023.114300","article-title":"Fructooligosaccharides attenuate non-alcoholic fatty liver disease by remodeling gut microbiota and association with lipid metabolism","volume":"159","author":"Huang","year":"2023","journal-title":"Biomed. Pharmacother."},{"key":"B27","doi-asserted-by":"publisher","first-page":"1992","DOI":"10.3390\/microorganisms9091992","article-title":"Comparative genomics and specific functional characteristics analysis of Lactobacillus acidophilus","volume":"9","author":"Huang","year":"2021","journal-title":"Microorganisms"},{"key":"B28","doi-asserted-by":"publisher","first-page":"4423","DOI":"10.1128\/JCM.40.12.4423-4427.2002","article-title":"Quantification of bacteria adherent to gastrointestinal mucosa by real-time PCR","volume":"40","author":"Huijsdens","year":"2002","journal-title":"J. Clin. Microbiol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1186\/s12986-016-0067-0","article-title":"Probiotics in prevention and treatment of obesity: a critical view","volume":"13","author":"Kobyliak","year":"2016","journal-title":"Nutr. Metab."},{"key":"B30","doi-asserted-by":"publisher","first-page":"158","DOI":"10.1016\/j.jff.2014.12.044","article-title":"Influences of structures of galactooligosaccharides and fructooligosaccharides on the fermentation in vitro by human intestinal microbiota","volume":"13","author":"Li","year":"2015","journal-title":"J. Funct. Foods"},{"key":"B31","doi-asserted-by":"publisher","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2013\u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"B32","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1111\/1462-2920.13589","article-title":"Formation of propionate and butyrate by the human colonic microbiota","volume":"19","author":"Louis","year":"2017","journal-title":"Environ. Microbiol."},{"key":"B33","doi-asserted-by":"publisher","first-page":"227","DOI":"10.3892\/ijmm.2025.5668","article-title":"Metabolic dysfunction-associated steatotic liver disease: pathogenesis, model and treatment","volume":"56","author":"Ma","year":"2025","journal-title":"Int. J. Mol. Med."},{"key":"B34","doi-asserted-by":"publisher","first-page":"1021","DOI":"10.3390\/nu9091021","article-title":"Effects of probiotics, prebiotics, and synbiotics on human health","volume":"9","author":"Markowiak","year":"2017","journal-title":"Nutrients"},{"key":"B35","doi-asserted-by":"publisher","first-page":"1394","DOI":"10.1136\/gut.2010.222844","article-title":"Hepatic fatty acid translocase CD36 upregulation is associated with insulin resistance, hyperinsulinaemia and increased steatosis in non-alcoholic steatohepatitis and chronic hepatitis C","volume":"60","author":"Miquilena-Colina","year":"2011","journal-title":"Gut"},{"key":"B36","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1080\/19490976.2015.1134082","article-title":"Formation of short chain fatty acids by the gut microbiota and their impact on human metabolism","volume":"7","author":"Morrison","year":"2016","journal-title":"Gut Microbes"},{"key":"B37","doi-asserted-by":"publisher","first-page":"111560","DOI":"10.1016\/j.biopha.2021.111560","article-title":"Probiotics-rich emulsion improves insulin signalling in Palmitate\/Oleate-challenged human hepatocarcinoma cells through the modulation of Fetuin-A\/TLR4-JNK-NF-\u03baB pathway","volume":"139","author":"Mularczyk","year":"2021","journal-title":"Biomed. Pharmacother."},{"key":"B38","doi-asserted-by":"publisher","first-page":"542","DOI":"10.3390\/nu12020542","article-title":"Beneficial effects of Lactobacillus plantarum strains on non-alcoholic fatty liver disease in high fat\/high fructose diet-fed rats","volume":"12","author":"Park","year":"2020","journal-title":"Nutrients"},{"key":"B39","doi-asserted-by":"publisher","first-page":"5613","DOI":"10.1007\/s00253-015-6482-8","article-title":"Evaluation of the probiotic properties of new Lactobacillus and Bifidobacterium strains and their in vitro effect","volume":"99","author":"Presti","year":"2015","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"802","DOI":"10.1038\/s41419-020-03003-w","article-title":"Understanding lipotoxicity in NAFLD pathogenesis: is CD36 a key driver?","volume":"11","author":"Rada","year":"2020","journal-title":"Cell Death Dis."},{"key":"B41","doi-asserted-by":"publisher","first-page":"103549","DOI":"10.1016\/j.isci.2021.103549","article-title":"In vitro models for non-alcoholic fatty liver disease: emerging platforms and their applications","volume":"25","author":"Ramos","year":"2022","journal-title":"Iscience"},{"key":"B42","doi-asserted-by":"publisher","first-page":"605","DOI":"10.1038\/s41575-019-0173-3","article-title":"Probiotics and prebiotics in intestinal health and disease: from biology to the clinic","volume":"16","author":"Sanders","year":"2019","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"1753","DOI":"10.1128\/AEM.01151-06","article-title":"Identification of prebiotic fructooligosaccharide metabolism in Lactobacillus plantarum WCFS1 through microarrays","volume":"73","author":"Saulnier","year":"2007","journal-title":"Appl. Environ. Microbiol."},{"key":"B44","doi-asserted-by":"publisher","first-page":"1772","DOI":"10.1093\/gbe\/evu137","article-title":"From prediction to function using evolutionary genomics: human-specific ecotypes of Lactobacillus reuteri have diverse probiotic functions","volume":"6","author":"Spinler","year":"2014","journal-title":"Genome Biol. Evol."},{"key":"B45","doi-asserted-by":"publisher","first-page":"958","DOI":"10.3390\/nu16070958","article-title":"Lactiplantibacillus plantarum ZDY2013 inhibits the development of non-alcoholic fatty liver disease by regulating the intestinal microbiota and modulating the PI3K\/Akt pathway","volume":"16","author":"Teng","year":"2024","journal-title":"Nutrients"},{"key":"B46","doi-asserted-by":"publisher","first-page":"D609","DOI":"10.1093\/nar\/gkae1010","article-title":"UniProt: the universal protein knowledgebase in 2025","volume":"53","year":"2025","journal-title":"Nucleic Acids Res."},{"key":"B47","doi-asserted-by":"publisher","first-page":"2035","DOI":"10.1136\/gutjnl-2016-312729","article-title":"Gut microbiome and liver diseases","volume":"65","author":"Tilg","year":"2016","journal-title":"Gut"},{"key":"B48","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1007\/s00394-018-1790-2","article-title":"Adipose tissue inflammation and metabolic syndrome. The proactive role of probiotics","volume":"58","author":"Torres","year":"2019","journal-title":"Eur. J. Nutr."},{"key":"B49","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1038\/s41575-018-0011-z","article-title":"The gut\u2013liver axis and the intersection with the microbiome","volume":"15","author":"Tripathi","year":"2018","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"B50","doi-asserted-by":"publisher","first-page":"305","DOI":"10.1007\/978-3-319-16104-4_27","article-title":"\u201cThe simulator of the human intestinal microbial ecosystem (SHIME\u00ae),\u201d","author":"Van de Wiele","year":"2015","journal-title":"The Impact of Food Bioactives on Health: In Vitro and Ex Vivo Models"},{"key":"B51","doi-asserted-by":"publisher","first-page":"103666","DOI":"10.1016\/j.jff.2019.103666","article-title":"Impact of probiotics and prebiotics targeting metabolic syndrome","volume":"64","author":"Xavier-Santos","year":"2020","journal-title":"J. Funct. Foods"},{"key":"B52","doi-asserted-by":"publisher","first-page":"105329","DOI":"10.1016\/j.jff.2022.105329","article-title":"In vitro fermentation of fructooligosaccharide and galactooligosaccharide and their effects on gut microbiota and SCFAs in infants","volume":"99","author":"Yao","year":"2022","journal-title":"J. Funct. Foods"},{"key":"B53","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1038\/nrgastro.2017.109","article-title":"Global burden of NAFLD and NASH: trends, predictions, risk factors and prevention","volume":"15","author":"Younossi","year":"2018","journal-title":"Nat. Rev. Gastroenterol. Hepatol."}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1866842\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T15:47:49Z","timestamp":1788968869000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1866842\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,9]]},"references-count":53,"alternative-id":["10.3389\/fmicb.2026.1866842"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2026.1866842","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,9]]},"article-number":"1866842"}}