Zion, O., Institute of Animal Science, ARO, Volcani Center, Bet Dagan 50250, Israel
Genin, O., Institute of Animal Science, ARO, Volcani Center, Bet Dagan 50250, Israel
Kawada, N., Department of Hepatology, Graduate School of Medicine, Osaka City University, Japan
Yoshizato, K., Department of Biological Science, Graduate School of Science, Hiroshima University, Japan
Roffe, S., Department of Animal Sciences, Hebrew University of Jerusalem, Rehovot, Israel
Nagler, A., Department of Hematology and Bone Marrow Transplantation, Chaim Sheba Medical Center, Tel Hashomer, Israel
Iovanna, J.L., INSERM U624, Stress Cellulaire, Campus de Luminy, Marseille, France
Halevy, O., Department of Animal Sciences, Hebrew University of Jerusalem, Rehovot, Israel
Pines, M., Institute of Animal Science, ARO, Volcani Center, Bet Dagan 50250, Israel

OBJECTIVES: Chronic pancreatitis is characterized by inflammation and fibrosis. We evaluated the efficacy of halofuginone, an inhibitor of collagen synthesis and myofibroblast activation, in preventing cerulein-induced pancreas fibrosis. METHODS: Collagen synthesis was evaluated by in situ hybridization and staining. Levels of prolyl 4-hydroxylase β (P4Hβ), cytoglobin/stellate cell activation-associated protein (Cygb/STAP), transgelin, tissue inhibitors of metalloproteinases, serum response factor, transforming growth factor β (TGFβ), Smad3, and pancreatitis-associated protein 1 (PAP-1) were determined by immunohistochemistry. Metalloproteinase activity was evaluated by zymography. RESULTS: Halofuginone prevented cerulein-dependent increase in collagen synthesis, collagen cross-linking enzyme P4Hβ, Cygb/STAP, and tissue inhibitors of metalloproteinase 2. Halofuginone did not affect TGFβ levels in cerulein-treated mice but inhibited serum response factor synthesis and Smad3 phosphorylation. In culture, halofuginone inhibited pancreatic stellate cell (PSC) proliferation and TGFβ-dependent increase in Cygb/STAP and transgelin synthesis and metalloproteinase 2 activity. Halofuginone increased c-Jun N-terminal kinase phosphorylation in PSCs derived from cerulein-treated mice. Halofuginone prevented the increase in acinar cell proliferation and further increased the cerulein-dependent PAP-1 synthesis. CONCLUSIONS: Halofuginone inhibits Smad3 phosphorylation and increases c-Jun N-terminal kinase phosphorylation, leading to the inhibition of PSC activation and consequent prevention of fibrosis. Halofuginone increased the synthesis of PAP-1, which further reduces pancreas fibrosis. Thus, halofuginone might serve as a novel therapy for pancreas fibrosis. © 2009 Lippincott Williams & Wilkins, Inc.