Effect of pro-inflammatory mediators on membrane-associated mucins expressed by human ocular surface epithelial cells
Effect of pro-inflammatory mediators on membrane-associated mucins expressed by human ocular surface epithelial cells
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DOI：
10.1016/j.exer.2009.12.009
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发表时间：
2010-03-01
影响因子：
3.4
通讯作者：
Gipson, Ilene K.
中科院分区：
文献类型：
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作者：
Albertsmeyer, Ann-Christin;Kakkassery, Vinodh;Gipson, Ilene K.
在非杂货干眼的眼部表面上，与膜相关的粘蛋白改变了。这项研究试图确定在干眼症患者的泪液中存在炎症介质是否在基因表达，蛋白质生物合成和/或外生域释放的水平下调节膜相关的粘蛋白MUC1和-16。使用了产生与膜相关的粘蛋白的人角膜边缘上皮细胞系（HCLE）。用白介素（IL）-6，-8或-17处理细胞，肿瘤坏死因子-Alpha（TNF-Alpha）和干扰素-GAMMA（IFN-GAMMA）或TNF-ALPHA和IFN-GAMMA的组合，或IFN-gamma和IL-17，为1、6、24或48 h。通过RT-PCR验证了这些介体的受体的存在。细胞因子对MUC1和-16表达水平的影响通过实时PCR，分别通过免疫印迹分析分别在细胞裂解物和培养基中分别在细胞裂解物和培养基中的粘蛋白蛋白蛋白生物合成和外生域释放。 TNF-Alpha和IFN-Gamma均显着诱导MUC1表达，细胞蛋白含量和外胞域随时间释放。与两种细胞因子的联合治疗不是添加剂。相比之下，炎症介质之一。 IFN-GAMMA影响了所有三个参数 - 基因表达，细胞蛋白和exodobain释放的MUC16。与TNF-Alpha和IFN-GAMMA的联合处理显示出类似于IFN-GAMMA的作用，除了外生域释放遵循TNF-Alpha的释放，TNF-Alpha诱导MUC16胞外域释放。总之，干眼症患者的泪液中存在的炎症介质会影响角膜上皮细胞上的MUC1和-16，并可能导致干眼症中表面粘液的改变。 （c）2009 Elsevier Ltd.保留所有权利。
Membrane-associated mucins are altered on the ocular surface in non-Sjogren's dry eye. This study sought to determine if inflammatory mediators, present in tears of dry eye patients, regulate membrane-associated mucins MUC1 and -16 at the level of gene expression, protein biosynthesis and/or ectodomain release. A human corneal limbal epithelial cell line (HCLE), which produces membrane-associated mucins, was used. Cells were treated with interleukin (IL)-6, -8, or -17, tumor necrosis factor-alpha (TNF-alpha), and Interferon-gamma (IFN-gamma), or a combination of TNF-alpha and IFN-gamma, or IFN-gamma and IL-17, for 1, 6, 24, or 48 h. Presence of receptors for these mediators was verified by RT-PCR. Effects of the cytokines on expression levels of MUC1 and -16 were determined by real-time PCR, and on mucin protein biosynthesis and ectodomain release in cell lysates and culture media, respectively, by immunoblot analysis. TNF-alpha and IFN-gamma each significantly induced MUC1 expression, cellular protein content and ectodomain release over time. Combined treatment with the two cytokines was not additive. By comparison, one of the inflammatory mediators. IFN-gamma, affected all three parameters-gene expression, cellular protein, and ectodomain release-for MUC16. Combined treatment with TNF-alpha and IFN-gamma showed effects similar to IFN-gamma alone, except that ectodomain release followed that of TNF-alpha, which induced MUC16 ectodomain release. In conclusion, inflammatory mediators present in tears of dry eye patients can affect MUC1 and -16 on corneal epithelial cells and may be responsible for alterations of surface mucins in dry eye. (C) 2009 Elsevier Ltd. All rights reserved.