Figure legend
Fig.1. Description and analysis of GP (A)Summarized extraction procedure. (B) HPLC of monosaccharide PMP derivatization. 1.PMP. 2. Mannose. 3. Rhamnose. 4. Glucuronic acid. 5.Galacturonic acid. 6. Glucose. 7. Galactose. 8. Arabinose. 9. Fucose. (C) Infrared spectroscopy. (D) Morphology by SEM. (E) Molecular weight distribution.
Fig. 2. GP alleviated the intestinal inflammation in rats with DSS induced colitis. (B)H&E
stained sections of the differently treated rats as described above. (C) (D) The effects of GP on Rats’ basal body weight and Spleen weight. (E) (F) and (G) Changes of cytokines in different groups in vivo. All data shown are representative of 3 independent experiments. Bars in graphs represent mean ± SD, # P<0.05,## P<0.01 VS Control group; *P<0.05, **P<0.01 VS DSS group.
Fig. 3. Effects of GP on gut microbiota. (A) Rarefaction curves of OTU quantity. (B)Heatmap at the level of phylumgenus. (C)Heatmap at the level of genus. (D) A phylogenetictree of OTUs by GraPhlAn visualization. (E) PCA analysis. (F) LPS in colon contents.
Fig. 4. GP inhibits inflammation via TLR4 pathway and activates mTOR dependent autophagy in DSS-induced colitis in vivo . (A) (C) Immunofluorescence for TLR4 and p62 was performed on colon sections. (B) (D) The expressions of TLR4 and p62 were calculated relative to DAPI staining from three independent experiments. (E) The expression of autophagy-related and inflammation-related proteins measured by western blot. (F) The expressions of proteins were quantified by the ratio of phosphorylated protein/total protein and total amount protein/GAPDH. All data shown are representative of 3 independent experiments. Bars in graphs represent mean ± SD, # P<0.05,## P<0.01 VS Control group; *P<0.05, **P<0.01 VS DSS group.
Fig. 5. Spearman correlation analysis of the bacterial genera with highest abundance and phenotypes.
Fig. 6. Transplanting of fecal microbiome intervened by GP improves intestinal inflammation. (A) The experimental protocol for FMT. (B) Changes of body weight (n=10). (C) Histopathological changes after HE staining. (D) Rarefaction curves of OUT quantity. (E) PCA analysis. (F) The gut microbiota composition among experimental groups at phylum/genus level.
Fig. 7. Transplanting of fecal microbiome intervened by GP improves intestinal inflammation. (A) Immunofluorescence for p-NF-κB was performed on colon sections. (B) The expression of autophagy-related and inflammation-related proteins measured by western blot. (C) (D) The expressions of proteins were quantified by the ratio of phosphorylated protein/total protein and total amount protein/GAPDH. All data shown are representative of 3 independent experiments. Bars in graphs represent mean ± SD, # P<0.05, ## P<0.01 VS Control group; *P<0.05, **P<0.01 VS DSS group.
Fig. 8. Blocking upstream TLR4-MAPK pathway activated autophagy and quenched inflammation. (A) Changes of cytokines contents in different groups in LPS-induced HT-29 cells. (B) The changes of autophagy-related and inflammation-related proteins measured by western blot. (C-G) The expressions of proteins were quantified by the ratio of phosphorylated protein/total protein and total amount protein/GAPDH. All data shown are representative of 3 independent experiments. Bars in graphs represent mean ± SD, *P<0.05, **P<0.01.
Fig. 9. Schematic representation of the proposed mechanism of GP on ameliorating intestinal inflammation.