Bourcier T, Forgez P, Borderie V, Scheer S, Rostène W, Laroche L
Cornea Bank, AP-HP, Paris VI University, the. Institut National de la Santé et de la Recherche Médicale (U33g), Saint-Antoine Hospital, Paris, France.
Invest Ophthalmol Vis Sci. 2000 Dec;41(13):4133-41.
To investigate whether human corneal epithelial cells express the glucocorticoid receptor (GR) and to assess the influence of dexamethasone (DEX) on these cells.
Human corneal epithelial cells 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-sulfop henyl) -2H-tetrazolium inner salt (MTS) assay at 2, 4, and 6 days of culture. Apoptosis was studied by nucleus labeling using a fluorescent dye and immunostaining by APO 2.7 at 6 days of culture. GR mRNA was detected in corneal epithelium and cultured corneal epithelial cells by means of reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemical staining of the epithelial cells was performed with a monoclonal anti-human GR.
RT-PCR and immunocytochemistry showed the expression of GR (mRNA and protein) in corneal epithelial cells. DEX significantly increased corneal epithelial cell proliferation with concentrations ranging from 10(-10) to 10(-6) M, with a maximum effect at 10(-7) M (P < 0.005). However, DEX also induced apoptosis of cultured corneal epithelial cells at any concentration used.
These results indicate that human corneal epithelial cells express the GR and proliferate in response to DEX stimulation which also induces corneal epithelial cell apoptosis.
研究人角膜上皮细胞是否表达糖皮质激素受体（GR），并评估地塞米松（DEX）对这些细胞的影响。
将人角膜上皮细胞培养于添加不同浓度DEX（范围为10⁻¹⁰至10⁻⁴ M）的培养基中。在培养2天、4天和6天时，通过3-(4,5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺基苯基)-2H-四唑内盐（MTS）法分析细胞增殖情况。在培养6天时，通过使用荧光染料进行细胞核标记和APO 2.7免疫染色研究细胞凋亡。通过逆转录-聚合酶链反应（RT-PCR）检测角膜上皮和培养的角膜上皮细胞中的GR mRNA。用抗人GR单克隆抗体对上皮细胞进行免疫细胞化学染色。
RT-PCR和免疫细胞化学显示角膜上皮细胞中存在GR（mRNA和蛋白质）表达。DEX在浓度范围为10⁻¹⁰至10⁻⁶ M时显著增加角膜上皮细胞增殖，在10⁻⁷ M时作用最强（P < 0.005）。然而，DEX在所用的任何浓度下也诱导培养的角膜上皮细胞凋亡。
这些结果表明人角膜上皮细胞表达GR，并在DEX刺激下增殖，同时DEX也诱导角膜上皮细胞凋亡。
