Figure 4
NOD1 expression in pancreatic acinar cells is necessary for the development of chronic pancreatitis induced by a low-dose cerulein and NOD1 ligand. Irradiated GFP-transgenic (GFP-Tg) NOD1-intact mice and NOD1-deficient mice were reconstituted with bone marrow (BM) cells from NOD1-deficient mice and GFP-Tg mice, respectively. These mice were referred to as NOD1−/−>GFP Tg and GFPTg>NOD1−/−, respectively. NOD1−/−>GFP Tg mice and GFPTg>NOD1−/− mice were administered a low-dose cerulein for a total of three times, or FK565 followed by a low-dose cerulein for a total of three times as described in Figure 1. Mice received each regimen twice a week for a total of 11 times and then sera and pancreatic tissues were obtained. Total numbers of mice in each group were as follows; NOD1−/−>GFP Tg cerulein (CER): n=6, NOD1−/−>GFP Tg CER+FK565: n=5, GFPTg>NOD1−/− CER: n=5, GFPTg>NOD1−/− CER+FK565: n=9. (a) Serum levels of amylase, pathological scores of the pancreas, and the numbers of pancreatic α-SMA+ cells per high-power fields obtained from mice three hours after the last injection of cerulein. Results are expressed as mean±s.e.m. and are a pool of three independent experiments. *P<0.05, **P<0.01 as compared with cerulein alone. (b) Concentrations of cytokines and chemokines in the serum were determined by ELISA. Results are expressed as mean±s.e.m. and are a pool of three independent experiments. **P<0.01 as compared with NOD1−/−>GFP Tg mice treated with cerulein alone, ##P<0.01 as compared with GFPTg>NOD1−/− mice treated with cerulein and FK565. (c) Representative picture of the pancreas tissue stained with H&E, anti-fibronectin Ab, anti-SMA Ab or anti-IL-33 Ab magnification × 400 (top four lines). Representative picture of dual immunofluorescence of the pancreas tissue stained with anti-GFP Ab (green color) or anti-pIκBα Ab (red color) or anti-pStat3 Ab (red color). Nuclei were stained with DAPI, magnification × 800 (bottom two lines). GFP, green fluorescent protein; H&E; hematoxylin and eosin; NOD1, nucleotide-binding oligomerization domain 1.
