藥物對體外培養之鞏膜纖維細胞的作用
Date Issued
2002
Date
2002
Author(s)
陳偉勵
DOI
902314B002444
Abstract
PURPOSE: 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.
Publisher
臺北市：國立臺灣大學醫學院眼科
Type
journal article
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