Galectin-8 mediates fibrogenesis induced by cyclosporine in human gingival fibroblasts.
fibrosis
focal adhesion kinase 1
galectins
gingival overgrowth
integrins
Journal
Journal of periodontal research
ISSN: 1600-0765
Titre abrégé: J Periodontal Res
Pays: United States
ID NLM: 0055107
Informations de publication
Date de publication:
Oct 2020
Oct 2020
Historique:
received:
11
11
2019
revised:
03
04
2020
accepted:
17
04
2020
pubmed:
26
5
2020
medline:
15
12
2020
entrez:
26
5
2020
Statut:
ppublish
Résumé
During cyclosporine-induced gingival overgrowth, the homeostatic balance of gingival connective tissue is disrupted leading to fibrosis. Galectins are glycan-binding proteins that can modulate a variety of cellular processes including fibrosis in several organs. Here, we study the role of galectin-8 (Gal-8) in the response of gingival connective tissue cells to cyclosporine. We used human gingival fibroblasts and mouse NIH3T3 cells treated with recombinant Gal-8 and/or cyclosporine for analyzing specific mRNA and protein levels through immunoblot, real-time polymerase chain reaction, ELISA and immunofluorescence, pull-down with Gal-8-Sepharose for Gal-8-to-cell surface glycoprotein interactions, short hairpin RNA for Gal-8 silencing and Student's t test and ANOVA for statistical analysis. Galectin-8 stimulated type I collagen and fibronectin protein levels and potentiated CTGF protein levels in TGF-β1-stimulated human gingival fibroblasts. Gal-8 interacted with α5β1-integrin and type II TGF-β receptor. Gal-8 stimulated fibronectin protein and mRNA levels, and this response was dependent on FAK activity but not Smad2/3 signaling. Cyclosporine and tumor necrosis factor alpha (TNF-α) increased Gal-8 protein levels. Finally, silencing of galectin-8 in NIH3T3 cells abolished cyclosporine-induced fibronectin protein levels. Taken together, these results reveal for the first time Gal-8 as a fibrogenic stimulus exerted through β1-integrin/FAK pathways in human gingival fibroblasts, which can be triggered by cyclosporine. Further studies should explore the involvement of Gal-8 in human gingival tissues and its role in drug-induced gingival overgrowth.
Sections du résumé
BACKGROUND AND OBJECTIVE
OBJECTIVE
During cyclosporine-induced gingival overgrowth, the homeostatic balance of gingival connective tissue is disrupted leading to fibrosis. Galectins are glycan-binding proteins that can modulate a variety of cellular processes including fibrosis in several organs. Here, we study the role of galectin-8 (Gal-8) in the response of gingival connective tissue cells to cyclosporine.
METHODS
METHODS
We used human gingival fibroblasts and mouse NIH3T3 cells treated with recombinant Gal-8 and/or cyclosporine for analyzing specific mRNA and protein levels through immunoblot, real-time polymerase chain reaction, ELISA and immunofluorescence, pull-down with Gal-8-Sepharose for Gal-8-to-cell surface glycoprotein interactions, short hairpin RNA for Gal-8 silencing and Student's t test and ANOVA for statistical analysis.
RESULTS
RESULTS
Galectin-8 stimulated type I collagen and fibronectin protein levels and potentiated CTGF protein levels in TGF-β1-stimulated human gingival fibroblasts. Gal-8 interacted with α5β1-integrin and type II TGF-β receptor. Gal-8 stimulated fibronectin protein and mRNA levels, and this response was dependent on FAK activity but not Smad2/3 signaling. Cyclosporine and tumor necrosis factor alpha (TNF-α) increased Gal-8 protein levels. Finally, silencing of galectin-8 in NIH3T3 cells abolished cyclosporine-induced fibronectin protein levels.
CONCLUSION
CONCLUSIONS
Taken together, these results reveal for the first time Gal-8 as a fibrogenic stimulus exerted through β1-integrin/FAK pathways in human gingival fibroblasts, which can be triggered by cyclosporine. Further studies should explore the involvement of Gal-8 in human gingival tissues and its role in drug-induced gingival overgrowth.
Substances chimiques
Galectins
0
Cyclosporine
83HN0GTJ6D
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
724-733Subventions
Organisme : Fondo Nacional de Desarrollo Científico y Tecnológico
ID : 1131122
Organisme : Fondo Nacional de Desarrollo Científico y Tecnológico
ID : 1170555
Informations de copyright
© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
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