Chen Wei-Li, Lin Chung-Tien, Yao Chung-Chen, Huang Yu-Hua, Chou Yu-Bin, Yin Hsiang-Shu, Hu Fung-Rong
Department of Ophthalmology and Graduate Institute of Clinical Medicine, National Taipei University Hospital, Taipei, Taiwan.
Ocul Immunol Inflamm. 2006 Aug;14(4):215-23. doi: 10.1080/09273940600732380.
To assess the in-vitro effects of dexamethasone (DEX) on the proliferation, apoptosis, and Na+-K+-ATPase activity of bovine corneal endothelial cells.
Bovine corneal endothelial cells were cultured with DEX ranging from 10-10 to 10-3 M. The effect of DEX on the proliferation was analyzed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay. Apoptosis and necrosis were detected by staining with fluorescein-conjugated annexin V and propidium iodide, followed by flow cytometry. The effect of DEX on Na+-K+-ATPase activity was evaluated using non-isotopic methods.
DEX did not affect cellular proliferation or induce apoptosis/necrosis from 10-10 to 10-5 M. At 10-4 and 10-3 M, DEX significantly decreased proliferation and increased apoptosis and/or necrosis. DEX significantly increased the Na+-K+-ATPase activity from 10-8 to 10-6 M, with the maximal effect at 10-6 M (p < 0.01); this effect was inhibited by RU38486, an antiglucocorticoid molecule.
Bovine corneal endothelial cells express glucocorticoid receptor (GR) mRNA and protein. DEX decreases cell proliferation and induces cellular apoptosis and/or necrosis at high concentrations. DEX also increases the Na+-K+-ATPase activity at certain concentrations.
评估地塞米松（DEX）对牛角膜内皮细胞增殖、凋亡及钠钾ATP酶活性的体外作用。
将牛角膜内皮细胞与浓度范围为10⁻¹⁰至10⁻³ M的DEX共同培养。通过3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺基苯基)-2H-四唑内盐（MTS）法分析DEX对增殖的影响。用荧光素偶联的膜联蛋白V和碘化丙啶染色，随后进行流式细胞术检测凋亡和坏死情况。采用非同位素方法评估DEX对钠钾ATP酶活性的影响。
在10⁻¹⁰至10⁻⁵ M浓度范围内，DEX不影响细胞增殖，也不诱导凋亡/坏死。在10⁻⁴和10⁻³ M时，DEX显著降低增殖并增加凋亡和/或坏死。在10⁻⁸至10⁻⁶ M浓度范围内，DEX显著增加钠钾ATP酶活性，在10⁻⁶ M时作用最大（p < 0.01）；该作用被抗糖皮质激素分子RU38486抑制。
牛角膜内皮细胞表达糖皮质激素受体（GR）mRNA和蛋白。高浓度时，DEX降低细胞增殖并诱导细胞凋亡和/或坏死。DEX在一定浓度下还会增加钠钾ATP酶活性。
