Fig. 4

Quantitative proteomic and biochemical analyses identify the differentially regulated proteins by anemoside B4 upon TNBS-induced colitis colon tissue from rats. Rats were orally administrated with 80 mg/kg TNBS. Anemoside B4 was continuously injected i.p at the doses of 10 mg/kg twice a day for 8 days. Rats were sacrificed on day 8 after colitis induction. a The volcano plot for the MS identified 604 proteins in colon tissue. Each point showed the log₂ (Fold change) in the x axis vs. their corresponding -log₁₀ (P-value) in the y axis. The filter boundaries were set so that a fold change above 1.5, or a fold change below 0.67, were considered significant, and a P-value of 0.05 was used as the cutoff value. The horizontal dotted line indicates the cutoff P-value of 0.05 and all points above this line have P < 0.05. The black points indicate 591 quantified unaltered proteins. The green points and red points indicated the down-regulated and up-regulated proteins, respectively. The circle indicated the proteins used for the biochemical validation. b The biological processes of the differentially regulated proteins analyzed by the online STRING database. c Quantification of S100A9 mRNA expression. d Western blotting analysis of S100A9 protein level. c, d Data were shown as mean ± SD (n = 3). ^##: P < 0.001 vs. vehicle group; ***: P < 0.001 vs. TNBS group. (E) Proteins in the downstream or upstream of S100A9 signaling pathway were detected by Western blotting in colon tissues (n = 3). f Analysis of S100A9 protein level. ^#: P < 0.05, ^##: P < 0.01, ^###: P < 0.001 vs. control group. **: P < 0.01 vs. TNBS group