DIFFERENTIAL PROTEIN EXPRESSION IN KERATOCONUS STROMAL FIBROBLAST

Abstract

Purpose: Keratoconus is a corneal disease characterized by a progressive thinning of the cornea, giving rise to a cone-shaped cornea, myopia, and irregular astigmatism. It has been considered the leading cause of corneal transplantation but although its importance, the exact pathologicals mechanisms associated are still unknown. The aim of this study was to analyze differential protein expression in keratoconus stromal fibroblast in comparison to healthy cornea stromal fibroblast.

Methods: Keratoconus and healthy cornea stromal fibroblasts were isolated, cultivated and characterizated by flow cytometry. After the proteins were extracted, purified a 2D electrophoresis were run and protein expression analysis were performed.

Results: There were no differences in morphology of fibroblasts from keratoconus and healthy cornea. They were positive to vimentine marker. The differential protein expression analysis of the keratoconus stromal fibroblast revealed differences in the protein expression profiles. Seven proteins were found downregulated and two proteins upregulated in the keratoconus stromal fibroblast. Nine proteins were only found in the healthy cornea stromal fibroblast and twenty six were only found in the stromal fibroblast from keratoconus. Identification of proteins by mass spectrometry is being performed.

Conclusions: A novel stromal primary culture of keratoconus fibroblasts was obtained and it was studied by proteomic view. Results showed differential protein expression in keratoconus stromal fibroblast in comparison to healthy cornea stromal fibroblasts. It can help us to understand the pathogenesis of keratoconus and may help us to find biomarkers for the prognosis and opportune diagnosis of keratoconus.

Keywords: 574 keratoconus • 663 proteomics • 484 cornea: stroma and keratocytes