Fig. 3: Blautia wexlerae-derived metabolites showed anti-inflammatory and anti-adipogenesis properties in adipocytes as well as alteration of mitochondrial metabolism.

A 3T3L1 pre-adipocytes were differentiated into mature adipocytes in the absence (none) or presence of the cultured supernatant of B. wexlerae at a final concentration of 10%. Gene expression of Pparγ (a transcription factor used as a marker of adipocyte differentiation), S100a8 (a chemokine for recruiting macrophages), and Nrf1 (a transcriptional factor used as a marker of mitochondrial biogenesis) was measured in 3T3L1 pre-adipocytes and adipocytes. Data are representative of two independent experiments without samples below the detection limit (n = 3–4, mean ± 1 SD). *P < 0.05; **P < 0.01; n.s. not significant (one-way ANOVA). B Mitochondrial mass was measured by flow cytometry analysis using Mitogreen in 3T3L1 adipocytes treated without or with culture supernatant (sup.) of B. wexlerae at a final concentration of 1% or 10%. Data are representative of two independent experiments (n = 4, mean ± 1 SD). **P = 0.0012 (one-way ANOVA). C By using an XF24 extracellular flux analyzer, the oxygen consumption rate (OCR) was measured in 3T3L1 adipocytes treated without or with the cultured supernatant of B. wexlerae at the concentration of 10%. Data are combined from two independent experiments (n = 14, mean ± 1 SD). D Lipid accumulation was assessed by using oil red O staining in 3T3L1 adipocytes treated without or with culture supernatant of B. wexlerae at a final concentration of 1% or 10%. Data are representative of two independent experiments (n = 4, mean ± 1 SD). *P = 0.0316 (one-way ANOVA). E Gene expression of Nrf1 in the eAT MAF of mice. Data are combined from two independent experiments without samples below the detection limit (n = 9–10, mean ± 1 SD). *P = 0.0421; **P < 0.0001 (one-way ANOVA). F Representative metabolites of glycolysis (lactate) and the TCA cycle (citrate, isocitrate, and succinate) in the eAT MAF of mice were measured by using liquid chromatography–tandem mass spectroscopy (LC–MS/MS). Data are combined from two independent experiments (n = 8, mean ± 1 SD). *P < 0.05; **P < 0.01 (one-way ANOVA). G Acetyl-l-carnitine, a constituent of the inner mitochondrial membrane, in the eAT MAF of mice was measured by using LC–MS/MS. Data are combined from two independent experiments without samples below the detection limit (n = 5–6, mean ± 1 SD). *P = 0.0231 (one-way ANOVA). CD CD-fed mice; HFD HFD-fed mice, HFD + Bw HFD-fed mice supplemented with B. wexlerae.