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.