Abstract
Purpose: :
The remodeling of scleral matrix is very important to investigate the pathogenesis and to develop therapeutic agents in various ocular diseases, including scleritis, orbital inflammatory diseases and vitreo–retinal diseases. In this study we generated the cell line from scleral stromal cells of a human eye and investigated the molecular mechanism of extracelluar matrix (ECM) remodelin.
Methods: :
This study has been approved by the Ethical Committee of Yamagata University Faculty of Medicine. After securing the written permission from the subject who underwent the enucleation due to malignant melanoma, the scleral stromal tissue was separated and cultured in DMEM with 10% fetal bovine serum, 37°C, CO2 5%. To establish the cell line, Human papilloma virus 16 E6 and E7 gene was transferred into the primary culture cells, and monoclonal cell lines were separated. The effects of platelet–derived growth factor–BB (PDGF–BB: 10ng/ml) and/or transforming growth factor–beta1 (TGF–beta1: 10ng/ml) on the expression of hyaluronan synthase (HAS)1,2,3, matrix metalloproteinase (MMP)1,2,3, tissue inhibitor of metalloproteinase (TIMP) 1,2, collagen type 1 ,3. The expression was observed by RT–PCR.
Results: :
PDGF–BB induced the expression level of MMP1, MMP3 and HAS2 at mRNA level. TGF–beta1 did not affect the expression of the investigated genes. The cross–talk between PDGF–BB and TGF–beta1 was not observed.
Conclusions: :
PDGF–BB stimulated the MMP1, MMP2 and HAS2 expression, which suggests that PDGF–BB is related to the scleral stromal remodeling.
Keywords: sclera • cytokines/chemokines • signal transduction