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.
To validate the protective effect of GP on intestinal inflammation associated with microbe-TLR4-autophagy axis, we detected the levels of several key proteins using Western blot analysis. FMT had almost no effects on the TLR4 pathway and mTOR-dependent autophagy, as shown in Fig. 7. Meanwhile, the colitis rats transplanted with DSS-treated microbiota showed a marked activation of the classic TLR4-p38 MAPK-NF-κB inflammatory pathway (Fig. 7A and B) as well as the phosphorylation of mTOR. Subsequently, the accumulation of p62 and the decrease of LCBII/LC3BI suggested defective autophagy. Notably, there was significant attenuation of TLR4 and degradation of p62 after FMT with GP-treated microbiota, which achieved similar therapeutic effects compared to GP lavage. In summary, the potential mechanism of GP on ameliorating intestinal inflammation via regulation of the gut microbiota and which was associated with TLR4-autophagy.