多功能隐形眼镜护理液及其成分对人眼角膜上皮细胞膜相关粘蛋白的影响。

Effects of multipurpose contact lens care solutions and their ingredients on membrane-associated mucins of human corneal epithelial cells.

机构信息

Department of Anatomy, Nagoya City University Medical School, Nagoya, Aichi, Japan.

出版信息

Eye Contact Lens. 2010 Nov;36(6):361-6. doi: 10.1097/ICL.0b013e3181faa43e.

DOI:10.1097/ICL.0b013e3181faa43e

PMID:21060261

Abstract

PURPOSE

Membrane-associated mucins play an important role for protecting epithelial cells at the ocular surface from microbial invasion. The purpose of this study was to determine whether multipurpose solutions (MPSs) for contact lens care and their ingredients alter the expression of membrane-associated mucins in human corneal epithelial (HCE-T) cells.

METHODS

SV40-immortalized HCE-T cells were cultured in Dulbecco's modified Eagle medium/F12 medium with 5% fetal bovine serum to confluence and were then exposed to 10% dilutions of five different MPSs, or to their representative ingredients, 0.1% macrogolglycerol hydroxystearate, 0.1% poloxamer, 0.1% poloxamine, 1 and 5 ppm polyhexamethylene biguanide, or 0.05% and 0.1% boric acid for 24 hr. Quantitative real-time polymerase chain reaction was used to investigate the gene expression of MUC1, MUC4, and MUC16. Immunofluorescence staining of MUC16 protein on the surface of the HCE-T cells exposed to 10% diluted MPSs for 24 hr or undiluted MPSs for 30 min was observed by laser confocal scanning microscopy followed by quantitative image analysis.

RESULTS

Three MPSs containing boric acid significantly reduced gene expressions of MUC1 from 20.2% to 56.7% (P<0.01). Gene expressions of MUC4 and MUC16 were also reduced by these MPSs; however, there were no significant differences. Among ingredients, 0.1% boric acid significantly reduced gene expressions of MUC1 and MUC16 by 7.4% and 18.9%, respectively (P<0.01). Immunofluorescence microscopy also demonstrated that in undiluted form, three MPSs containing boric acid significantly reduced the expression of MUC16 protein.

CONCLUSIONS

The MPSs containing boric acid downregulate membrane-associated mucins as compared with MPSs that do not contain boric acid. There may be some subtle membrane or other interactions between ingredients in lens-care solutions that adversely alter corneal cell mucins.

摘要

目的

膜相关粘蛋白在保护眼表面上皮细胞免受微生物入侵方面起着重要作用。本研究旨在确定接触镜护理用多功能溶液（MPS）及其成分是否会改变人角膜上皮（HCE-T）细胞中膜相关粘蛋白的表达。

方法

SV40 永生化 HCE-T 细胞在含有 5%胎牛血清的 Dulbecco 改良 Eagle 培养基/F12 培养基中培养至汇合，然后用 5 种不同 MPS 的 10%稀释液或其代表性成分 0.1%聚乙二醇甘油羟硬脂酸酯、0.1%泊洛沙姆、0.1%泊洛胺、1 和 5ppm 聚六亚甲基双胍或 0.05%和 0.1%硼酸暴露 24 小时。实时定量聚合酶链反应用于研究 MUC1、MUC4 和 MUC16 的基因表达。用激光共聚焦扫描显微镜观察暴露于 10%稀释 MPS 24 小时或未稀释 MPS 30 分钟的 HCE-T 细胞表面 MUC16 蛋白的免疫荧光染色，并进行定量图像分析。

结果

三种含有硼酸的 MPS 显著降低了 MUC1 的基因表达，从 20.2%降至 56.7%（P<0.01）。这些 MPS 还降低了 MUC4 和 MUC16 的基因表达，但无统计学差异。在成分中，0.1%硼酸显著降低了 MUC1 和 MUC16 的基因表达，分别降低了 7.4%和 18.9%（P<0.01）。免疫荧光显微镜也表明，在未稀释的形式下，三种含有硼酸的 MPS 显著降低了 MUC16 蛋白的表达。