title: Inhibition of transforming growth factor beta signaling by halofuginone as a modality for pancreas fibrosis prevention.
abstract: OBJECTIVE
  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 beta (P4Hbeta), cytoglobin/stellate cell activation-associated protein (Cygb/STAP), transgelin, tissue inhibitors of metalloproteinases, serum response factor, transforming growth factor beta (TGFbeta), 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 P4Hbeta, Cygb/STAP, and tissue inhibitors of metalloproteinase 2. Halofuginone did not affect TGFbeta levels in cerulein-treated mice but inhibited serum response factor synthesis and Smad3 phosphorylation. In culture, halofuginone inhibited pancreatic stellate cell (PSC) proliferation and TGFbeta-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.
authors.affiliation: Institute of Animal Sciences, The Volcani Center, Bet Dagan, Israel.
authors.forename: Orit
authors.initials: O
authors.lastname: Zion
authors.forename: Olga
authors.initials: O
authors.lastname: Genin
authors.forename: Norifumi
authors.initials: N
authors.lastname: Kawada
authors.forename: Katsutoshi
authors.initials: K
authors.lastname: Yoshizato
authors.forename: Suzy
authors.initials: S
authors.lastname: Roffe
authors.forename: Arnon
authors.initials: A
authors.lastname: Nagler
authors.forename: Juan L
authors.initials: JL
authors.lastname: Iovanna
authors.forename: Orna
authors.initials: O
authors.lastname: Halevy
authors.forename: Mark
authors.initials: M
authors.lastname: Pines
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002108:Ceruletide; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D000078842:Cytoglobin; D005109:Extracellular Matrix / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005355:Fibrosis / Q000139:chemically induced / Q000517:prevention & control; D005914:Globins / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007274:Injections, Intraperitoneal; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010179:Pancreas / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D000073718:Pancreatitis-Associated Proteins; D010880:Piperidines / Q000008:administration & dosage / Q000494:pharmacology*; D011393:Procollagen-Proline Dioxygenase / Q000378:metabolism; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000008:administration & dosage / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism; D052999:Quinazolinones / Q000008:administration & dosage / Q000494:pharmacology*; D026362:Serum Response Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D051900:Smad3 Protein / Q000378:metabolism; D019714:Tissue Inhibitor of Metalloproteinases / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: C000627265:CYGB protein, human; C000627267:Cygb protein, mouse; D000078842:Cytoglobin; D000073718:Pancreatitis-Associated Proteins; D010880:Piperidines; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D052999:Quinazolinones; C000612141:REG3A protein, human; C490253:Reg3b protein, mouse; D026362:Serum Response Factor; D051900:Smad3 Protein; C497791:Smad3 protein, mouse; D019714:Tissue Inhibitor of Metalloproteinases; D016212:Transforming Growth Factor beta; D002108:Ceruletide; D005914:Globins; D003094:Collagen; D011393:Procollagen-Proline Dioxygenase; D048049:Extracellular Signal-Regulated MAP Kinases; C526500:P4hb protein, mouse; D019704:Protein Disulfide-Isomerases; C010176:halofuginone