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Invest Ophthalmol Vis Sci. 2020 Mar 9;61(3):14. doi: 10.1167/iovs.61.3.14.
We used a human corneal epithelial cell (HCE) line to determine the involvement of the advanced glycation end products (AGEs) / receptor for AGEs (RAGE) couple in corneal epithelium wound healing.
After wounding, HCE cells were exposed to two major RAGE ligands (HMGB1 and AGEs), and wound healing was evaluated using the in vitro scratch assay. Following wound healing, the HCE cells were used to study the influence of the RAGE ligands on HCE proliferation, invasion, and migration. Activation of the nuclear factor (NF)-κB signaling pathway by the AGEs/RAGE couple was tested using a luciferase reporter assay. Functional transcriptional regulation by this pathway was confirmed by quantification of expression of the connexin 43 target gene. For each experiment, specific RAGE involvement was confirmed by small interfering RNA treatments.
AGEs treatment at a dose of 100 µg/mL significantly improved the wound healing process in a RAGE-dependent manner by promoting cell migration, whereas HMGB1 had no effect. No significant influence of the AGEs/RAGE couple was observed on cell proliferation and invasion. However, this treatment induced an early activation of the NF-κB pathway and positively regulated the expression of the target gene, connexin 43, at both the mRNA and protein levels.
Our results demonstrate that the RAGE pathway is activated by AGEs treatment and is involved in the promotion of corneal epithelial wound healing. This positive action is observed only during the early stages of wound healing, as illustrated by the quick activation of the NF-κB pathway and induction of connexin 43 expression.
我们使用人角膜上皮细胞（HCE）系来确定晚期糖基化终产物（AGEs）/AGEs 受体（RAGE）对角膜上皮伤口愈合的影响。
在创伤后，HCE 细胞暴露于两种主要的 RAGE 配体（HMGB1 和 AGEs），并通过体外划痕试验评估伤口愈合情况。在伤口愈合后，使用 HCE 细胞研究 RAGE 配体对 HCE 增殖、侵袭和迁移的影响。通过荧光素酶报告基因测定检测 AGEs/RAGE 对核因子（NF）-κB 信号通路的激活作用。通过定量连接蛋白 43 靶基因的表达来确认该途径的功能转录调控。对于每个实验，通过小干扰 RNA 处理来确认特定的 RAGE 参与。
100μg/ml 的 AGEs 处理以 RAGE 依赖性方式显著改善了伤口愈合过程，促进了细胞迁移，而 HMGB1 则没有作用。AGEs/RAGE 对细胞增殖和侵袭没有显著影响。然而，这种处理诱导了 NF-κB 途径的早期激活，并正向调节了靶基因连接蛋白 43 的表达，mRNA 和蛋白质水平均上调。
我们的结果表明，RAGE 途径被 AGEs 处理激活，并参与促进角膜上皮伤口愈合。这种积极作用仅在伤口愈合的早期阶段观察到，如 NF-κB 途径的快速激活和连接蛋白 43 表达的诱导所示。
