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A recent study demonstrated that 6-week oral supplementation of Anaerostipes rhamnosivorans (ARHAM) combined with the prebiotic myo-inositol (MI) reduced fasting glucose levels in mice. In the present study, we investigated the effects of a 13-week ARHAM-MI supplementation in high-fat diet-fed mice and examined the metabolic fate of MI, including its microbial conversion into short-chain fatty acids (SCFAs), using 13C-MI and stable isotope tracers in the cecum, portal vein, and peripheral blood. The results showed that the ARHAM-MI group gained less weight than the MI-only and placebo groups. Analysis of intestinal mRNA and stable isotope tracing revealed that MI is primarily absorbed in the upper gastrointestinal tract, whereas microbial conversion to SCFAs predominantly occurs in the cecum and is enhanced by ARHAM. ARHAM-MI mice also showed increased cecal Gpr43 mRNA expression, indicating enhanced SCFA-mediated signaling. Notably, SCFAs derived from MI displayed distinct distribution patterns: 13C-butyrate was detected exclusively in the cecum, 13C-propionate was present in the cecum and portal vein, whereas 13C-acetate was the only SCFA detected in peripheral blood. Collectively, ARHAM-MI co-supplementation confers modest metabolic benefits in high-fat diet-fed mice, underscoring the need to optimize the dosage and administration frequency of ARHAM-MI to enhance its therapeutic efficacy.<\/jats:p>","DOI":"10.3390\/ijms26199340","type":"journal-article","created":{"date-parts":[[2025,9,25]],"date-time":"2025-09-25T08:56:44Z","timestamp":1758790604000},"page":"9340","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Intestinal Myo-Inositol Metabolism and Metabolic Effects of Myo-Inositol Utilizing Anaerostipes rhamnosivorans in Mice"],"prefix":"10.3390","volume":"26","author":[{"given":"Aldo","family":"Grefhorst","sequence":"first","affiliation":[{"name":"Department of Internal and (Experimental) Vascular Medicine, Amsterdam University Medical Centers, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonella S.","family":"Kleemann","sequence":"additional","affiliation":[{"name":"Department of Internal and (Experimental) Vascular Medicine, Amsterdam University Medical Centers, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Havik","sequence":"additional","affiliation":[{"name":"Department of Internal and (Experimental) Vascular Medicine, Amsterdam University Medical Centers, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7635-5244","authenticated-orcid":false,"given":"Antonio Dario","family":"Troise","sequence":"additional","affiliation":[{"name":"Proteomics, Metabolomics & Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80055 Portici, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9249-7058","authenticated-orcid":false,"given":"Sabrina","family":"De Pascale","sequence":"additional","affiliation":[{"name":"Proteomics, Metabolomics & Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80055 Portici, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9362-8515","authenticated-orcid":false,"given":"Andrea","family":"Scaloni","sequence":"additional","affiliation":[{"name":"Proteomics, Metabolomics & Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80055 Portici, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Max","family":"Nieuwdorp","sequence":"additional","affiliation":[{"name":"Department of Internal and (Experimental) Vascular Medicine, Amsterdam University Medical Centers, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5366-5960","authenticated-orcid":false,"given":"Thi Phuong Nam","family":"Bui","sequence":"additional","affiliation":[{"name":"Department of Internal and (Experimental) Vascular Medicine, Amsterdam University Medical Centers, 1105 AZ Amsterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1038\/s41569-023-00847-5","article-title":"Obesity and the risk of cardiometabolic diseases","volume":"20","author":"Valenzuela","year":"2023","journal-title":"Nat. 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