Myungmoon Bio, Hwaseong, Korea.
Department of Physiology, College of Korean Medicine Dongguk University, Gyeongju, Korea.
Korean J Ophthalmol. 2020 Feb;34(1):1-10. doi: 10.3341/kjo.2019.0046.
Diquafosol is a pharmaceutical drug used for dry eye treatment with a novel mechanism of action. It is a purinergic P2Y2 receptor agonist that promotes the secretion of tears and healing of corneal epithelial wounds. However, its inhibitory effect on hyperosmotic stress-induced inflammation in human corneal epithelial cells (HCECs) remains unclear.
A hyperosmotic stress model was established by transferring HCECs from isosmotic (312 mOsm/kg to hyperosmotic medium (500 mOsm/kg). HCECs were incubated with 500 mOsm/kg hyperosmotic medium for 30 minutes, and then treated with diquafosol (0.6-6 mg/mL) for 4 or 24 hours. Cells were then harvested and analyzed by western blot, immunocytochemistry, and real-time polymerase chain reaction to evaluate the expression of interleukin-6, tumor necrosis factor-alpha, and the phosphorylation status of nuclear factor-kappa B.
Diquafosol significantly decreased the mRNA and protein expression of hyperosmotic stress-induced tumor necrosis factor-alpha and interleukin-6. These results were supported by immunofluorescence staining and quantitative real-time polymerase chain reaction analysis. Furthermore, diquafosol inhibits nuclear factor-kappa B activation by suppressing the phosphorylation and degradation of the inhibitor of кB.
This study shows that diquafosol inhibits nuclear factor-kappa B signaling and inflammatory factors induced by hyperosmotic stress in HCECs. This suggests that using diquafosol for the improvement of dry eye syndrome could be effective in the treatment of inflammation-related corneal and conjunctival diseases.
地夸磷索是一种用于治疗干眼症的药物，具有新颖的作用机制。它是一种嘌呤能 P2Y2 受体激动剂，可促进泪液分泌和角膜上皮伤口愈合。然而，其对人角膜上皮细胞（HCEC）中高渗应激诱导的炎症的抑制作用尚不清楚。
通过将 HCEC 从等渗（312 mOsm/kg）转移至高渗培养基（500 mOsm/kg）来建立高渗应激模型。将 HCEC 在 500 mOsm/kg 的高渗培养基中孵育 30 分钟，然后用地夸磷索（0.6-6 mg/mL）处理 4 或 24 小时。然后通过 Western blot、免疫细胞化学和实时聚合酶链反应分析收集细胞，以评估白细胞介素-6、肿瘤坏死因子-α和核因子-κB 的磷酸化状态的表达。
地夸磷索可显著降低高渗应激诱导的肿瘤坏死因子-α和白细胞介素-6 的 mRNA 和蛋白表达。免疫荧光染色和定量实时聚合酶链反应分析支持了这些结果。此外，地夸磷索通过抑制抑制剂 кB 的磷酸化和降解来抑制核因子-κB 的激活。
本研究表明地夸磷索可抑制 HCEC 中高渗应激诱导的核因子-κB 信号和炎症因子。这表明，使用地夸磷索改善干眼症综合征可能对治疗与炎症相关的角膜和结膜疾病有效。
