Mochizuki Hiroshi, Fukui Masaki, Hatou Shin, Yamada Masakazu, Tsubota Kazuo
Division for Vision Research, National Institute of Sensory Organs, National Tokyo Medical Center, Tokyo, Japan.
Clin Ophthalmol. 2010 Aug 19;4:925-30. doi: 10.2147/opth.s12648.
A glycocalyx plays important roles in the ocular surface epithelium, but there is no direct simple method to evaluate ocular surface glycocalyx. We tested a wheat germ agglutinin conjugate of fluorescein (F-WGA) as a marker to demonstrate ocular surface glycocalyx in vivo.
After a 5% F-WGA solution was applied to the eyes of eight healthy volunteers, fluorescent intensities of the central cornea and bulbar conjunctiva were measured by fluorophotometry. A 10% fluorescein-conjugated dextran solution served as the control. Changes in fluorescent intensities of the ocular surface following a challenge with 5% N-acetyl cysteine, a conventional mucolytic agent, were tested in the second experiment. Saline instillation served as a control.
The ocular surface was diffusely stained by F-WGA. Breakup of the precorneal tear film was not apparent, possibly because F-WGA was bound to the glycocalyx of the ocular surface epithelium. F-WGA fluorescent intensities were high in the superior, nasal, and inferior regions of the bulbar conjunctiva and low in the temporal conjunctiva and cornea. No such regional differences were observed with fluorescein-conjugated dextran. F-WGA fluorescent intensities decreased significantly with N-acetyl cysteine instillation, whereas they were not affected by saline instillation.
The fluorophotometric method may be used to evaluate the glycocalyx quantitatively in the ocular surface in vivo.
糖萼在眼表上皮中发挥着重要作用，但尚无直接简便的方法来评估眼表糖萼。我们测试了荧光素标记的麦胚凝集素（F-WGA）作为一种标记物，以在体内显示眼表糖萼。
将5%的F-WGA溶液滴入8名健康志愿者的眼中后，通过荧光光度法测量中央角膜和球结膜的荧光强度。10%的荧光素标记葡聚糖溶液作为对照。在第二个实验中，测试了用传统黏液溶解剂5%N-乙酰半胱氨酸刺激后眼表荧光强度的变化。滴注生理盐水作为对照。
眼表被F-WGA弥漫性染色。角膜前泪膜破裂不明显，可能是因为F-WGA与眼表上皮的糖萼结合。F-WGA荧光强度在球结膜的上、鼻和下区域较高，而在颞侧结膜和角膜中较低。荧光素标记葡聚糖未观察到这种区域差异。滴注N-乙酰半胱氨酸后，F-WGA荧光强度显著降低，而滴注生理盐水则对其无影响。
荧光光度法可用于在体内定量评估眼表的糖萼。
