Chapman University School of Pharmacy, Chapman University, Irvine, CA 92618, USA.
Int J Mol Sci. 2022 Nov 18;23(22):14289. doi: 10.3390/ijms232214289.
Diabetes mellitus causes several detrimental effects on the ocular surface, including compromised barrier function and an increased risk of infections. The glycocalyx plays a vital role in barrier function. The present study was designed to test the effect of a high glucose level on components of glycocalyx. Stratified human corneal and conjunctival epithelial cells were exposed to a high glucose concentration for 24 and 72 h. Changes in Mucin (MUC) 1, 4, 16 expression were quantified using real-time PCR and ELISA. Rose bengal and jacalin staining were used to assess the spatial distribution of MUC16 and O-glycosylation. Changes in the gene expression of five glycosyltransferases and forty-two proteins involved in cell proliferation and the cell cycle were also quantified using PCR and a gene array. High glucose exposure did not affect the level or spatial distribution of membrane-tethered MUC 1, 4, and 16 either in the corneal or conjunctival epithelial cells. No change in gene expression in glycosyltransferases was observed, but a decrease in the gene expression of proteins involved in cell proliferation and the cell cycle was observed. A high-glucose-mediated decrease in gene expression of proteins involved in cellular proliferation of corneal and conjunctival epithelial cells may be one of the mechanisms underlying a diabetes-associated decrease in ocular surface's glycocalyx.
糖尿病可对眼表面造成多种有害影响,包括损害屏障功能和增加感染风险。糖萼在屏障功能中起着至关重要的作用。本研究旨在测试高糖水平对糖萼成分的影响。将分层的人角膜和结膜上皮细胞暴露于高葡萄糖浓度下 24 和 72 小时。使用实时 PCR 和 ELISA 定量检测粘蛋白 (MUC) 1、4、16 的表达变化。使用玫瑰红 Bengal 和 jacalin 染色评估 MUC16 和 O-糖基化的空间分布。使用 PCR 和基因芯片还定量检测了 5 种糖基转移酶和 42 种参与细胞增殖和细胞周期的蛋白质的基因表达变化。高葡萄糖暴露对角膜或结膜上皮细胞中膜结合的 MUC 1、4 和 16 的水平或空间分布均无影响。未观察到糖基转移酶的基因表达发生变化,但观察到参与细胞增殖和细胞周期的蛋白质的基因表达下降。高葡萄糖介导的角膜和结膜上皮细胞中参与细胞增殖的蛋白质的基因表达下降可能是糖尿病相关眼表面糖萼减少的机制之一。