Abstract
Abstract: :
Purpose: To investigate whether bovine scleral fibroblasts express the glucocorticoid receptor (GR), and to access the influence of dexamethasone (DEX) on scleral fibroblasts. Methods: Bovine scleral fibroblasts were cultured in medium added with different concentrations of DEX ranging from 10-10 to 10-4 M. RU38486, an antiglucocorticoid molecule was used to inhibit the effects caused by DEX. Cell proliferation was analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide ) (MTT) assay at 1,3,5,7 days of culture. The apoptosis was studied by terminal deoxynucleotidyl transferase-mediated dUTP nicked-end labeling (TUNEL stain). Glucocorticoid receptor mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemical staining of the cells was performed with a monoclonal anti-human GR. Results: RT-PCT and immunocytochemistry showed the expression of GR (mRNA and protein) in cultured bovine scleral fibroblasts. Dexamethasone significantly increased scleral fibroblast's proliferations ranging from 10-8 to 10-5M, with a maximal effects at 10-6 M (P<0.005). There were nearly no viable cells in medium containing 10-4 M of DEX. The proliferative or toxic effect of DEX was inhibited by RU38486 in all culture conditions. Conclusion: Bovine scleral fibroblasts expressed GR. Different DEX concentrations may lead to proliferative or toxic effects of cultured bovine scleral fibroblasts.
Keywords: 574 sclera • 323 apoptosis/cell death • 377 corticosteroids