Effect of granulocyte macrophage-colony stimulating factor on extracellular matrix deposition by dermal fibroblasts from patients with scleroderma.

Abstract

OBJECTIVE:

. To investigate the in vitro effect of granulocyte macrophage-colony stimulating factor (GM-CSF) on the deposition of extracellular matrix (ECM) in fibroblasts obtained from the skin of patients with systemic sclerosis (Ssc), compared to healthy controls.

METHODS:

Dermal fibroblasts obtained from 14 patients with SSc (7 with the diffuse form and 7 with CREST syndrome) and from 7 controls were studied. Both SSc and normal skin fibroblast cultures were stimulated for 4 and 8 days with 100 ng/ml GM-CFS. GM-CSF receptor (GM-CSFR) expression was determined by Western blot of cell lysates. Immunofluorescence was used to determine GM-CSFR expression and to investigate the deposition of ECM (type I collagen, fibronectin, and tenascin). Quantitative analysis of ECM was performed by ELISA. Expression of type I collagen and metalloproteinase 1 (MMP-1) mRNA was determined by real-time quantitative PCR.

RESULTS:

Deposition of ECM by normal fibroblasts appeared not to be influenced by stimulation with GM-CSF; in contrast, after stimulation with GM-CSF SSc fibroblasts showed increased deposition of fibronectin and tenascin, while type I collagen production was decreased; these results were found with both immunofluorescence and ELISA. Quantitative PCR revealed that GM-CSF inhibited the expression of mRNA type I collagen in SSc fibroblasts but not in normal fibroblasts, whereas levels of the main collagenolytic enzyme, MMP-1, were not affected.

CONCLUSION:

These results suggest that in SSc fibroblasts GM-CSF exerts a blocking effect on the deposition of type I collagen, through an inhibitory action on mRNA, while the production of other components of ECM such as fibronectin and tenascin is increased by stimulation with this cytokine.

• Authors: Loredana Postiglione, Paolo Ladogana, Stefania Montagnani, Gaetano di Spigna, Clotilde Castaldo, Mimmo Turano, Eugenia Maria Bruno, Franca Di Meglio, Antonio Riccio, Guido Rossi
• Journal: The Journal of rheumatology (J Rheumatol) Vol. 32 Issue 4 Pg. 656-64 (Apr 2005) ISSN: 0315-162X [Print] Canada
• PMID: 15801021 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Collagen Type I
• Extracellular Matrix Proteins
• Fibronectins
• RNA, Messenger
• Receptors, Granulocyte-Macrophage Colony-Stimulating Factor
• Tenascin
• Granulocyte-Macrophage Colony-Stimulating Factor
• Matrix Metalloproteinase 1

Topics

• Adult
• Aged
• CREST Syndrome (metabolism, pathology)
• Cells, Cultured
• Collagen Type I (metabolism)
• Dermis (drug effects, metabolism, pathology)
• Dose-Response Relationship, Drug
• Extracellular Matrix (drug effects, metabolism)
• Extracellular Matrix Proteins (metabolism)
• Female
• Fibroblasts (drug effects, metabolism, pathology)
• Fibronectins (metabolism)
• Gene Expression (drug effects)
• Granulocyte-Macrophage Colony-Stimulating Factor (pharmacology)
• Humans
• Male
• Matrix Metalloproteinase 1 (genetics, metabolism)
• Middle Aged
• RNA, Messenger
• Receptors, Granulocyte-Macrophage Colony-Stimulating Factor (drug effects, metabolism)
• Scleroderma, Diffuse (metabolism, pathology)
• Tenascin (metabolism)