陳偉勵2006-07-262018-07-122006-07-262018-07-122002http://ntur.lib.ntu.edu.tw//handle/246246/26728PURPOSE: To investigate whether cultured bovine scleral fibroblasts express the glucocorticoid receptor (GR) and to assess the influence of dexamethasone (DEX) on these cells. METHODS: Bovine scleral fibroblasts were cultured in medium supplemented with various concentrations of DEX (ranging from 10(-10) to 10(-4) M). Cell proliferation was analyzed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-s ulfophenyl)-2H-tetrazolium inner salt (MTS) assay at 2, 4, and 6 days of culture. Some experiments were performed in the presence of mifepristone (RU38486), an antiglucocorticoid molecule. The early phase of apoptosis was studied by means of slceral fibroblast staining with a fluorescein conjugate of annexin V and propidium iodide, and cells were analyzed by flow cytometry. Glucocorticoid receptor mRNA was detected in keratocytes by means of reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemical staining of the cells was performed with a monoclonal anti-human GR. RESULTS: RT-PCR and immunocytochemistry showed no result of GR (mRNA and protein) in cultured bovine scleral fibroblast. The reason may be due to improper primer, and short length of bovine GR checked from Genebank. However, there existed positive staining of dexamethasone on scleral fibroblast by immunocytochemical staining. As for the proliferative response of Dexamethasone, it has no par ticular increased of decreased scleral proliferation with concentrations ranging from 10(-9) to 10(-5) M,. Dexamethasone's proproliferative effect was not inhibited by RU38486. However, DEX did induced some apoptosis of cultured bovine scleral fibroblasts at any concentration used.application/pdf184062 bytesapplication/pdfzh-TW國立臺灣大學醫學院眼科journal articlehttp://ntur.lib.ntu.edu.tw/bitstream/246246/26728/1/902314B002444.pdf