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

机译:多功能隐形眼镜护理液及其成分对人角膜上皮细胞膜相关黏蛋白的影响。

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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.
机译:目的:膜相关粘蛋白在保护眼表皮细胞免受微生物侵袭中起着重要作用。这项研究的目的是确定用于隐形眼镜护理的多功能溶液(MPS)及其成分是否会改变人角膜上皮(HCE-T)细胞中膜相关黏蛋白的表达。方法:将SV40永生化的HCE-T细胞在含5%胎牛血清的Dulbecco改良Eagle培养基/ F12培养基中培养至汇合,然后暴露于10%的五种不同MPS稀释液或它们的代表成分0.1%的聚乙二醇甘油羟基硬脂酸酯中,0.1%的泊洛沙姆,0.1%的泊洛沙明,1和5 ppm的聚六亚甲基双胍或0.05%和0.1%的硼酸处理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。通过激光共聚焦扫描显微镜,然后进行定量图像分析,观察了暴露于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。结果:3种含硼酸的MPS将MUC1的基因表达从20.2%降低至56.7%(P <0.01)。这些MPSs也降低了MUC4和MUC16的基因表达。但是,没有显着差异。在各成分中,0.1%的硼酸分别使MUC1和MUC16的基因表达分别降低7.4%和18.9%(P <0.01)。免疫荧光显微镜检查还表明,未经稀释的三种含硼酸的MPS明显降低了MUC16蛋白的表达。结论:与不含硼酸的MPS相比,含硼酸的MPS下调膜相关粘蛋白。镜片护理液中成分之间可能存在一些微妙的膜或其他相互作用,从而不利地改变了角膜细胞粘蛋白。

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