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. 2020 Nov 5;11(3):857–880. doi: 10.1016/j.jcmgh.2020.10.015

Figure 2.

Figure 2

H pylori promotes gastric epithelial cell senescence. (A) Gram staining of PMSS1. (B) CXCL8 mRNA was measured using qPCR after AGS cells were infected with PMSS1 for 8 hours. Means ± SD. Representative of 2 independent repeats analyzed by an unpaired t test. (C) NF-κB signaling was assessed using Western blot analysis after AGS cells were infected with PMSS1 for 8 hours. This process was repeated twice. (D) Schematic diagram of the H pylori–infected mouse model. (E) SA-β-gal staining (left upper panel) and IHC staining of H pylori colonization (left lower panel) in the H pylori–infected and control mouse groups (samples are from antrum). Right: The histology score of H pylori colonization is shown. Plot: median. The red arrow refers to typical H pylori staining (S shape). Scale bar: 100 μm (upper panel); 50 μm (lower panel). (F) Senescent cells were detected by SA-β-gal staining in mucosa samples from PMSS1-infected mice (n = 12). Quantification was performed according to pathologic types (the 4 diagnoses were defined according to the same strategy we used in Figure 1B; n = 12 for each category). Plot: median. Lower: Representative H&E images are shown. Black arrow: Typical goblet cells. Representative Alcian blue staining of goblet cells also is shown in the bottom right of the H&E image. Red arrow: Typical nuclear dysplasia. Scale bar: 100 μm. All images were taken from the antrum except that images about IM were from the junction between the antrum and corpus. (G) Representative human mucosa samples (from antrum) negative and positive for H pylori infection as detected using IHC. (H) Senescent cells detected by Sudan Black B in H pylori–negative and H pylori–positive human precancerous lesions (from antrum). Senescent cells were analyzed only in atrophic mucosa without IM or DP. Samples from 20 patients diagnosed with CG, AG, IM, and DP. n (negative) = 11, n (positive) = 9. Plot: median. Mann–Whitney test. (I and J) SA-β-gal staining and BrdU labeling in AGS and GES-1 cells. Cells were first exposed to 5 Gy radiation to induce senescence and then co-cultured with H pylori strain PMSS1 for 5 days (means ± SD, unpaired t test). (G–J) Scale bar: 50 μm. ∗P < .05. ∗∗P < .01. ∗∗∗∗P < .0001. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NFR, nuclear fast red; SBB, Sudan Black B.