In vitro P144 and P17 Anti-TGF-β Peptides Effect in TGFβ-Stimulated Corneal Epithelial Cells

Abstract

Purpose: : TGF-β is a pleiotropic molecule whose expression is increased in inflammatory, proliferative, and degenerative ocular pathologies. We have previously showed that TGF-β upregulated MMP-9, MMP-13, and some cytokine secretion by a human corneal epithelial cell line. P144 and P17 are two anti-TGF-β peptides that have showed to block the profibrogenic TGF-β activity in liver, heart, skin, and in SIRC cells. The aim of this research was to study the effect of P144 and P17 anti-TGF-β peptides in TGF-β-stimulated corneal epithelial cells in vitro.

Methods: : The human corneal epithelial (HCE) cell line was exposed to TGF-β1 and -β2 for 30min, 2h, 24h and 48h in the presence/absence of P144 and P17 anti-TGF-β peptides. pSMAD2 expression was determined by Western blot analysis and immunofluorescence assays. MMP-9 and -13 secretion was analyzed in 48h cell supernatants by inmunobead based assay in a Luminex IS-100. Cell viability was determined by XTT assay.

Results: : Both P144 and P17 peptides downregulated TGF-β1-stimulated MMP13 secretion. P144 and P17 did not inhibit TGFβ2-stimulated MMP13 secretion or TGF-β1 and -β2-stimulated MMP-9 secretion. pSMAD2 expression was decreased in the presence of P144 peptide but not by P17 peptide. There were not significant differences in cell viability between controls and P144- or P17-treated cells.

Conclusions: : P144 and/or P17 anti-TGF-β peptides could be a potential topical therapy for ocular surface inflammatory pathology as evidence suggests that they can downregulate some TGF-β-related events in epithelial cells.

Keywords: growth factors/growth factor receptors • cornea: epithelium