Trypsin activity governs increased susceptibility to pancreatitis in mice expressing human PRSS1R122H

Abstract

Figure 2. Transgenic expression of human PRSS1 R122H caused severe AP. (A) Schema of cerulein-induced AP protocol. (B) Representative photos of pancreata from transgenic human PRSS1 R122H mice and C57BL/6J mice 24 hours after cerulein induction (n = 8). (C) Significant increase in pancreatic edema (pancreas-to-body weight ratio) in PRSS1 R122H mice 24 hours after cerulein induction. Mean SEM (n = 8). *P < 0.05; ***P < 0.001; 2-way ANOVA with Tukey's test. (D) Serum amylase levels after 24 hours of cerulein induction. Mean SEM (n = 8). ***P < 0.001; 2-way ANOVA with Tukey's test. (E) Representative images of H&E staining of the pancreata (n = 8). Scale bars: 300 m. (F) Histology score evaluation of AP. Mean SEM (n = 8). ***P < 0.001; 2-way ANOVA with Tukey's test. (G) Representative immunohistochemical staining for CD11b (pan leukocytes), F4/80 (macrophage), and Gr-1 (neutrophil) positive inflammatory cells (brown signal with hematoxylin purple counterstain) on sections from transgenic PRSS1 R122H mice and C57BL/6J mice 24 hours after cerulein induction (n = 8). Scale bars: 200 m. (H) Immunohistochemical staining for analysis of p65 nuclear translocation, an indicator of NF-B activation in the pancreata of PRSS1 R122H mice and C57BL/6J mice (n = 8). Scale bars: 200 m. (I) Quantification of p65 nuclear translocation in the pancreata of PRSS1 R122H mice and C57BL/6J mice. Mean SEM (n = 8). ***P < 0.001; 2-way ANOVA with Tukey's test. (J) Pancreatic mRNA expression levels of monocyte chemoattractant protein-1 (Mcp1), tumor necrosis factor alpha (Tnfa), interleukin 1 (Il1b), and Il6 in PRSS1 R122H mice and C57BL/6J mice were measured by real-time RT-PCR. Mean SEM (n = 4). *P < 0.05; ***P < 0.001; 2-way ANOVA with Tukey's test.