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

Figure 6 Pristimerin upregulates p57 to modulate glycolytic reprogramming and ameliorate the spasmolytic polypeptide-expressing metaplasia phenotype in gastric organoids. A: Pristimerin upregulates gastric mucosal p57 in spasmolytic polypeptide-expressing metaplasia (SPEM) mice. Scale bar, 20 μm; B: Representative images of normal gastric organoids, N-methyl-N’-nitro-N-nitrosoguanidine/Helicobacter pylori-induced SPEM gastric organoids, and those following Cdkn1c overexpression or pristimerin intervention, along with quantitative analyses of organoid number and diameter. Scale bar, 100 μm; C: P57 expression in SPEM gastric organoids following Cdkn1c overexpression or pristimerin treatment. Scale bar, 10 μm; D-F: Both Cdkn1c overexpression and pristimerin treatment suppressed cell proliferation [reverse transcription quantitative polymerase chain reaction (RT-qPCR); Western blot, β-actin-normalized relative expression vs control] in SPEM gastric organoids, downregulated key glycolytic regulators (RT-qPCR; Western blot), stem cell markers (immunofluorescence: Scale bar, 10 μm), and SPEM-specific markers (RT-qPCR; Western blot), while promoting gastric intrinsic factor (RT-qPCR; Western blot) expression with pristimerin exhibiting stronger effects. ^aP < 0.05. ^bP < 0.01. ^cP < 0.001. ^dP < 0.0001. Values represent the mean ± SD (A: n = 6; B-F: n = 5). NS: Not significant; Cdkn1c-OE: Cdkn1c-overexpressing spasmolytic polypeptide-expressing metaplasia gastric organoids; 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; HIF-1α: Hypoxia-inducible factor-1α; LDHA: Lactate dehydrogenase A; PKM2: Pyruvate kinase M2; MUC6: Mucin 6; TFF2: Trefoil factor 2; WFDC2: Whey-acidic-protein four-disulfide core domain protein 2; Lgr5: Leucine-rich repeat-containing G-protein coupled receptor 5; Troy: Tumor necrosis factor receptor superfamily member 19.