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. 2024 Jan 18;14:34. doi: 10.1038/s41398-024-02755-9

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — A Water consumption of NT mice with food restriction (Veh: n = 4, Met: n = 4; p = 0.170; RM-ANOVA), AD mice with food restriction (Veh: n = 8, Met: n = 5; p = 0.160; mixed effects model), and AD mice without food restriction (Veh: n = 2, Met: n = 2; p = 0.696; RM-ANOVA). B Weight of NT mice with food restriction (Veh: n = 16, Met: n = 16; p = 0.634; mixed effects model), AD mice with food restriction (Veh: n = 14, Met: n = 12; p = 0.792; mixed effects model), and AD mice without food restriction (Veh: n = 6, Met: n = 5; p = 0.589; RM-ANOVA). C Fasting blood glucose concentration of NT mice with food restriction (Veh: n = 16, MET: n = 16; p = 0.996; mixed effects model), AD mice with food restriction (Veh: n = 14, Met: n = 12; p = 0.849; mixed effects model), and AD mice without food restriction (Veh: n = 6, Met: n = 5; p = 0.178; RM-ANOVA). D CNCbl and HOCbl concentration in the serum of NT and AD mice (n = 4 per group; CNCbl, NT mice, p = 0.113; AD mice, p = 0.615; HOCbl, NT mice, p = 0.397; AD mice, p = 0.321; t test). E Score scatter plot showing the distribution of vehicle- and metformin-treated NT and AD mice based on principal component analysis. F Permutational multivariate analysis of variance showing changes in serum metabolites of vehicle- and metformin-treated NT and AD mice clustered by group and genotype (genotype, r² = 0.272, p < 0.001; treatment, r² = 0.08, p = 0.204). G Heatmap of hierarchical clustering analysis showing chemically and biochemically classified metabolites of serum from vehicle- and metformin-treated NT and AD mice clustered by group and genotype. H Multiple linear regression analysis showing changes in serum metabolites of vehicle- and metformin-treated NT and AD mice. Full names of metabolites were shown in Table S2. I Metabolite set enrichment analysis showing changes in serum metabolites of vehicle- and metformin-treated groups of NT mice. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01 versus Veh. Veh, vehicle; Met metformin, NT non-transgenic C57BL/6 mice, AD 3xTg-AD mice, Res food restriction, CNCbl cyanocobalamin, HOCbl, hydroxocobalamin.