Disruption of Zonula Occludens-1 Localization in the Rabbit Corneal Epithelium by Contact Lens-Induced Hypoxia
Disruption of Zonula Occludens-1 Localization in the Rabbit Corneal Epithelium by Contact Lens-Induced Hypoxia
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DOI：
10.1167/iovs.09-3407
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发表时间：
2009-10-01
影响因子：
4.4
通讯作者：
Nishida, Teruo
中科院分区：
文献类型：
--
作者：
Yanai, Ryoji;Ko, Ji-Ae;Nishida, Teruo
目的.缺氧损害角膜上皮的屏障功能。这种功能依赖于紧密连接，其中闭锁小带（ZO）-1是主要成分。作者研究了缺氧对兔角膜上皮ZO-1定位和表达的影响。将聚甲基丙烯酸甲酯（PMMA）或刚性透气（RGP）透镜应用于白化病兔的每只眼睛24小时。通过活体共聚焦显微镜检查角膜上皮的结构，并通过测量中央角膜厚度来评价上皮屏障功能。ZO-1在角膜上皮中的分布和表达分别通过免疫荧光分析、免疫印迹和逆转录-聚合酶链反应分析进行检测。应用PMMA透镜，而不是RGP透镜，导致角膜上皮表面的细胞大小减少，与对照眼相比，中央角膜厚度增加。免疫荧光分析显示，在对照眼或用RGP透镜治疗的眼中，角膜上皮细胞周围的ZO-1免疫反应性呈连续模式。相比之下，在用PMMA透镜治疗的眼睛中，ZO-1染色的模式是不连续和斑片状的。PMMA透镜可降低角膜上皮ZO-1 mRNA和蛋白的表达，而RGP透镜则无此作用。眼表缺氧诱导兔角膜上皮细胞间紧密连接破坏。（Invest Ophthalmol维斯科学。2009; 50：4605-4610）DOI：10.1167/iovs.09-3407
PURPOSE. Hypoxia impairs the barrier function of the corneal epithelium. This function depends on tight junctions, of which zonula occludens (ZO)-1 is a major component. The authors have investigated the effects of hypoxia on ZO-1 localization and expression in the rabbit corneal epithelium in vivo.METHODS. A polymethylmethacrylate (PMMA) or rigid gas-permeable (RGP) lens was applied to one eye each of albino rabbits for 24 hours. The structure of the corneal epithelium was examined by in vivo confocal microscopy, and epithelial barrier function was evaluated by measurement of central corneal thickness. The distribution and expression of ZO-1 in the corneal epithelium were examined by immunofluorescence analysis and by immunoblot and reverse transcription-polymerase chain reaction analyses, respectively.RESULTS. Application of a PMMA lens, but not that of an RGP lens, resulted in a reduction in cell size at the surface of the corneal epithelium, compared with that in control eyes, and an increase in central corneal thickness. Immunofluorescence analysis revealed a continuous pattern of ZO-1 immunoreactivity around the perimeter of superficial corneal epithelial cells in control eyes or in eyes treated with an RGP lens. In contrast, the pattern of ZO-1 staining was discontinuous and patchy in eyes treated with a PMMA lens. Amounts of ZO-1 mRNA and protein in the corneal epithelium were reduced by application of a PMMA lens but not by that of an RGP lens.CONCLUSIONS. Hypoxia at the ocular surface induced the disruption of tight junctions between superficial cells in the rabbit corneal epithelium in vivo. (Invest Ophthalmol Vis Sci. 2009; 50: 4605-4610) DOI:10.1167/iovs.09-3407