Inhibition of transforming growth factor beta signaling by halofuginone as a modality for pancreas fibrosis prevention.
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.
Zion
Orit
O
Institute of Animal Sciences, The Volcani Center, Bet Dagan, Israel.
Genin
Olga
O
Kawada
Norifumi
N
Yoshizato
Katsutoshi
K
Roffe
Suzy
S
Nagler
Arnon
A
Iovanna
Juan L
JL
Halevy
Orna
O
Pines
Mark
M
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*
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