OH-CATH30 targets CD40 to suppress NF-κB signaling and promote arachidonic acid metabolic reprogramming in acute pancreatitis

Figure 5 OH-CATH30 inhibits CD40 expression and nuclear factor-kappa B activation in pancreatic acinar cells and acute pancreatitis mice. A-C: Western blotting analysis of CD40 and nuclear factor-kappa B (NF-κB) p65 protein expression in 266-6 cells. Representative immunoblots(A); densitometric quantification of CD40 (B) and NF-κB p65 (C) normalized to that of glyceraldehyde-3-phosphate dehydrogenase. Cells were treated with control, cholecystokinin (100 nM), or cholecystokinin + OH-CATH30 (100 nM); D-F: Western blotting analysis of CD40 and NF-κB p65 protein expression in mouse pancreatic tissues. Representative immunoblots(D); quantitative analysis of CD40 (E) and NF-κB p65 (F) protein levels normalized to that of glyceraldehyde-3-phosphate dehydrogenase. Mice were assigned to control, acute pancreatitis, or acute pancreatitis + OH-CATH30 (30 μg/kg) groups; G-I: Immunohistochemical staining of CD40 and NF-κB p65 in pancreatic tissue sections. Representative images showing CD40 and NF-κB p65 expression (brown staining) (G); scale bar = 50 μm. Quantitative analysis of integrated optical density for CD40 (H) and NF-κB p65 (I) staining. All data are presented as mean ± SD [n = 3 independent experiments (A-C) or n = 6 mice per group (D-I)]. Statistical analysis was performed using one-way ANOVA followed by Tukey’s HSD post hoc test. ^aP < 0.05 vs control, ^bP < 0.05 vs cholecystokinin or acute pancreatitis group. NF-κB: Nuclear factor-kappa B; AP: Acute pancreatitis; CCK: Cholecystokinin; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.