Pristimerin ameliorates spasmolytic polypeptide-expressing metaplasia by modulating Cdkn1c (p57)-mediated glycolytic reprogramming

Figure 3 Inhibitory effect of pristimerin on aberrantly expressed spasmolytic polypeptide-expressing metaplasia-specific markers and stem cell markers. A: Immunofluorescence co-localization analysis of chief cell marker gastric intrinsic factor with spasmolytic polypeptide-expressing metaplasia (SPEM)-specific markers trefoil factor 2 (TFF2)/mucin 6 (MUC6) in mouse gastric tissues, and detection of TFF2/MUC6 expression by reverse transcription quantitative polymerase chain reaction in mouse stomach. Scale bar, 20 μm; B: Expression of SPEM-specific marker whey-acidic-protein four-disulfide core domain protein 2 (WFDC2) in mouse gastric tissues. Scale bar, 100 μm; C: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay of human gastric epithelial (GES-1) cells viability treated with varying concentrations of tamoxifen or pristimerin; D: Western blot analysis of SPEM-specific markers (TFF2/MUC6/WFDC2) in GES-1 cells with β-actin-normalized quantification (vs control); E and F: Inhibitory effect of pristimerin on stem cell markers leucine-rich repeat-containing G-protein coupled receptor 5 and tumor necrosis factor receptor superfamily member 19 in gastric mucosa of SPEM mice (E) and GES-1 cells (F). Scale bar, 20 μm. ^aP < 0.05; ^bP < 0.01; ^cP < 0.001; ^dP < 0.0001. Values represent the mean ± SD (A, B and E: n = 6; C, D and F: n = 5). DAPI: 4’,6-diamidino-2-phenylindole; GIF: Gastric intrinsic factor; Pri-L: Low-dose pristimerin; Pri-M: Medium-dose pristimerin; Pri-H: High-dose pristimerin; IOD: Integrated optical density; MUC6: Mucin 6; TAM: Tamoxifen; TFF2: Trefoil factor 2; WFDC2: Whey-acidic-protein four-disulfide core domain protein 2; MOD: Mean optical density; Lgr5: Leucine-rich repeat-containing G-protein coupled receptor 5; Troy: Tumor necrosis factor receptor superfamily member 19.