Fig 1: LGALS9 RNA expression was elevated in platelets from SAP mice. (A and B) Upregulated genes in platelets from SAP mouse models at 24 h and 168 h. (C) 414 commonly upregulated genes identified in both 24 h and 168 h groups. (D) Functional enrichment analysis of the common upregulated genes was performed using GO and KEGG. (E) LGALS9 RNA expression in platelets from the NC, 24 h, and 168 h groups. All data represented the means ± SEM. n=5. *P < 0.05, ***P < 0.001, two-sided Student’s t-test.
Fig 2: LGALS9 suppressed SAP inflammation and NLRP3-dependent pyroptosis in vivo. (A and B) Western Blot analysis of the expression levels of pyroptosis-related proteins in pancreatic tissues from each group, n=3. (C) ELISA detection of IL-1β, IL-18, IL-6, and TNF-α levels in serum from each group, n=5. Means ± SEM; NS, not significant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 as determined by two-sided Student’s t-test.
Fig 3: LGALS9 alleviated pancreatic tissue inflammation and injury in SAP in vivo. (A) Representative images of H&E staining and MPO immunohistochemical staining in each group (Scale bar: 100 μm). (B) Pancreatic histology scores in each group. (C) Percentage of MPO-positive area in each group. (D and E) Representative images and results of TUNEL staining in pancreatic tissue from each group (Scale bar: 100 μm). Means ± SEM. n=5. NS, not significant, *P < 0.05, ***P < 0.001, ****P < 0.0001 as determined by two-sided Student’s t-test.
Fig 4: LGALS9 inhibited inflammation and NLRP3-dependent pyroptosis in vitro. (A and B) Representative images and results of calcein (green), PI (red), and DAPI (blue) staining in mouse pancreatic acinar cells from each group (Scale bar: 100 μm). (C) ELISA detection of IL-1β, IL-18, IL-6, and TNF-α levels in cell culture supernatants from each group. (D–G) Western Blot analysis of the expression levels of pyroptosis-related proteins, including ASC, NLRP3, caspase1-FL, caspase1-cleaved, GSDMD-FL, and GSDMD-N expression in acinar cells from each group. Means ± SEM. n=3. NS, not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, two-sided Student’s t-test.
Fig 5: LGALS9 protein expression was elevated in platelets and serum and correlated with pyroptosis-related proteins. (A and B) Expression levels of LGALS9 in platelet releasates and serum from SAP patients compared with NC, n=6. (C and D) Expression levels of LGALS9 in platelet releasates and serum from SAP mice compared with NC, n=5. (E and F) Correlations between LGALS9 and pyroptosis-related proteins, including NLRP3, PYCARD (ASC), CASP1 (caspase-1), GSDMD, IL1B (IL-1β), and IL-18 in humans and mice. (G) Correlation analysis of LGALS9 with pyroptosis-related proteins using the mRNA expression profile dataset GSE194331. Means ± SEM; *P < 0.05, **P < 0.01, ****P < 0.0001 as determined by two-sided Student’s t-test.
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