Granulocyte colony-stimulating factor reduces fibrosis in a mouse model of chronic pancreatitis

PLoS One. 2014 Dec 31;9(12):e116229. doi: 10.1371/journal.pone.0116229. eCollection 2014.

Abstract

Background: Chronic pancreatitis (CP) is a necroinflammatory process resulting in extensive pancreatic fibrosis. Granulocyte colony-stimulating factor (G-CSF), a hematopoietic stem cell mobilizer, has been shown to exert an anti-fibrotic effect partly through the enrichment of bone marrow (BM) cells in fibrotic organ. We aimed to test the effect of G-CSF on fibrosis in a mouse model of CP.

Methods: CP was induced in C57Bl/6J mice by consecutive cerulein injection (50 µg/kg/day, 2 days a week) for 6 weeks. Mice were then treated with G-CSF (200 µg/kg/day, 5 day a week) or normal saline for 1 week, and sacrificed at week 7 or week 9 after first cerulein injection. Pancreatic histology, pancreatic matrix metallopeptidase 9 (MMP-9), MMP-13 and collagen expression were examined. Pancreatic myofibroblasts were isolated and cultured with G-CSF. Collagen, MMP-9 and MMP-13 expression by myofibroblasts was examined. The BM-mismatched mice model was used to examine the change of BM-derived myofibroblasts and non-myofibroblastic BM cells by G-CSF in the pancreas.

Results: The pancreatic collagen expression were significantly decreased in the G-CSF-treated group sacrificed at week 9. While collagen produced from myofibroblasts was not affected by G-CSF, the increase of MMP13 expression was observed in vitro. There were no effect of G-CSF in the number of myofibroblasts and BM-derived myofibroblasts. However, the number of non-myofibroblastic BM cells and macrophages were significantly increased in the pancreata of cerulein- and G-CSF-treated mice, suggesting a potential anti-fibrotic role of non-myofibroblastic BM cells and macrophages stimulated by G-CSF.

Conclusions: Our data indicated that G-CSF contributed to the regression of pancreatic fibrosis. The anti-fibrotic effects were possibly through the stimulation of MMP-13 from myofibroblasts, and the enhanced accumulation of non-myofibroblastic BM cells and macrophages in the pancreas.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Marrow Cells / drug effects
  • Cell Movement / drug effects
  • Cells, Cultured
  • Collagen / metabolism
  • Disease Models, Animal
  • Female
  • Fibrosis / drug therapy
  • Fibrosis / pathology
  • Granulocyte Colony-Stimulating Factor / pharmacology*
  • Green Fluorescent Proteins / genetics
  • Macrophages / drug effects
  • Male
  • Matrix Metalloproteinase 13 / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Myofibroblasts / drug effects
  • Myofibroblasts / metabolism
  • Pancreas / drug effects
  • Pancreas / metabolism
  • Pancreatitis, Chronic / drug therapy*
  • Pancreatitis, Chronic / metabolism
  • Pancreatitis, Chronic / pathology*

Substances

  • Granulocyte Colony-Stimulating Factor
  • Green Fluorescent Proteins
  • Collagen
  • Matrix Metalloproteinase 13
  • Matrix Metalloproteinase 9

Grants and funding

This study was funded by grants from Chang Gung Memorial Hospital, Taiwan (CMRPG3B047, CMRPG3A092), Ministry of Science and Technology (100-2314-B-182A-056, 101-2314-B-182-096) and The New Century Health Care Promotion Foundation, Taiwan (IACUC NO:2012081402). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.