Fig 2: Fourteen days of calorie restriction decreased the bacterial antigen load and systemic inflammatory marker in mice. a Serum LBP. n = 10 for the NC group and n = 6 for the CR group. b Serum TNF-a. n = 10 for both groups. Data are shown as the mean ± s.e.m. Unpaired t test (two-tailed) was used to analyse variation between the NC and CR groups. *P < 0.05 and ***P < 0.001

Fig 3: Effects of Atractylodes macrocephala polysaccharide, chlorogenic acid, and geniposide combination on lipopolysaccharide signaling and endotoxemia in non-alcoholic steatohepatitis induced by high-fat diet. (A) Immunofluorescence staining for F4/80 in the liver tissue (×600 magnification). (B) The LPS level in the plasma, the hepatic LBP content, and the mRNA expression of CD14 and MyD88 in the liver tissue. (C) The hepatic content of IL-1ß and TNF-a. CTR, control; HFD, high-fat diet; ACG, combination consisting of Atractylodes macrocephala polysaccharide, chlorogenic acid, and geniposide; NaB, sodium butyrate; LPS, lipopolysaccharide; LBP, lipopolysaccharide-binding protein; IL-1ß, interleukin-1ß; TNF-a, tumor necrosis factor–a. *p < 0.05 and **p < 0.01.

Fig 4: L. murinus CR147 reduces intestinal permeability and systemic inflammatory marker in old microbiota-colonized gnotobiotic mice. a Intestinal permeability of mice colonized with old microbiota (OM) or OM plus L. murinus CR147 (OM + CR147) measured by FITC-dextran translocation to systemic circulation following oral gavage (OM, n = 7; OM + CR147, n = 6). b Serum LBP (OM, n = 7; OM + CR147, n = 6). c Serum TNF-a (n = 5 per group). All data are shown as the mean ± s.e.m. Unpaired t test (two-tailed) was used to analyse the variation between the two groups. *P < 0.05 and **P < 0.01

Fig 5: Protection on intestinal barrier function contributes to the therapeutic effects of GC combination on NASH. Overgrowth of the Gram-negative bacteria leads to increased content of LPS in the intestine and gut barrier dysfunction. LPS activates RhoA/ROCK signaling which induces activation of MLC2 via direct phosphorylation or phosphorylation of MYPT. The increased phosphorylation of MLC2 leads to the rearrangement of TJ proteins and increase of TJ permeability. The gut-derived LPS go into liver via portal circulation and activate KCs to produce inflammatory factors participating in the onset of NASH. GC combination was found to restore the TJs protein expression and down-regulate RhoA/ROCK signaling while ameliorated NASH. GC, Geniposide and chlorogenic acid combination, IL-1ß, interleukin-1ß, LBP, lipopolysaccharide binding protein, LPS, lipopolysaccharide, MLC, myosin light chain, MYPT, myosin phosphatase target subunit, NASH, non-alcoholic steatohepatitis, ROCK, Rho-associated kinase, TNF, tumor necrosis factor, TLR, Toll-like receptor, TJ, tight junction.