Seo Kyoung Yul, Chung So-Hyang, Lee Joon H, Park Mi Young, Kim Eung Kweon
Department of Ophthalmology, Institute of Vision Research, Yonsei University College of Medicine, Seoul, Korea.
Cornea. 2007 Jul;26(6):709-14. doi: 10.1097/ICO.0b013e31804f5a09.
To study the influence of dexamethasone on membrane-associated mucins produced by human corneal epithelial cells.
Human corneal epithelial cells were cultured in medium supplemented with various concentrations of dexamethasone (ranging from 10 to 10 M). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis using monoclonal antibodies specific for human MUC1 (HMFG-1), MUC4 (1G8), and MUC16 (OC125) were performed to evaluate the effect of dexamethasone on membrane-associated mucin expression. The effect of glucocorticoid receptor antagonist (RU38486) on dexamethasone-induced mucin expression was estimated.
RT-PCR revealed that MUC1 and MUC16 gene expression were upregulated 48 hours after addition of dexamethasone and that MUC4 gene expression was downregulated in the same condition. Western blot analysis showed that MUC1 and MUC16 proteins were increased after addition of dexamethasone. However, MUC4 was not detected by anti-MUC4 monoclonal antibody (1G8) for ASGP-2 under our conditions. Treatment with RU38486 inhibited the changes of MUC1, MUC4, and MUC16 by dexamethasone; thus, the effect of dexamethasone on mucin expression is mediated by glucocorticoid receptors.
This study shows that MUC1, MUC4, and MUC16 are regulated differently by dexamethasone in human corneal epithelial cells. External application of dexamethasone might affect the precorneal mucin.
研究地塞米松对人角膜上皮细胞产生的膜相关黏蛋白的影响。
将人角膜上皮细胞培养于添加不同浓度地塞米松(范围为10至10⁻⁶ M)的培养基中。采用逆转录聚合酶链反应(RT-PCR)以及使用针对人MUC1(HMFG-1)、MUC4(1G8)和MUC16(OC125)的单克隆抗体进行蛋白质印迹分析,以评估地塞米松对膜相关黏蛋白表达的影响。评估糖皮质激素受体拮抗剂(RU38486)对地塞米松诱导的黏蛋白表达的作用。
RT-PCR显示,添加地塞米松48小时后,MUC1和MUC16基因表达上调,而在相同条件下MUC4基因表达下调。蛋白质印迹分析表明,添加地塞米松后MUC1和MUC16蛋白增加。然而,在我们的条件下,抗MUC4单克隆抗体(1G8)未检测到ASGP-2的MUC4。用RU38486处理可抑制地塞米松引起的MUC1、MUC4和MUC16的变化;因此,地塞米松对黏蛋白表达的作用是由糖皮质激素受体介导的。
本研究表明,地塞米松在人角膜上皮细胞中对MUC1、MUC4和MUC16的调节方式不同。地塞米松的外用可能会影响角膜前黏蛋白。