Abstract

Purpose

To investigate the efficacy of topical application of 3% diquafosol sodium (DQS) and tocopherol (TCP) acetate mixtures in a mouse model of experimental dry eye (EDE).

Methods

After exposure to desiccating stress for 5 days, eye drops consisting of 3% DQS alone, 0.01% TCP alone, or 3% DQS and 0.005% or 0.01% TCP mixture were applied for the treatment of EDE. Tear volume, tear film break-up time (TBUT), corneal fluorescein staining scores (CFSS), and tear film lipid layer grades (TFLLG) were measured at 0, 5 and 10 days after treatment. The 2',7'-dichlorodihydrofluorescein diacetate assay (DCFDA) for reactive oxygen species (ROS) production, enzyme-linked immunosorbent assay (ELISA) for malondialdehyde (MDA), and flow cytometry for CD4 + interferon (IFN)-γ+ T cells were evaluated on the ocular surface at 10 days after treatment. In addition, levels of tumour necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and chemokine CC motif ligand 4 (CCL4) in the conjunctiva were measured using a multiplex immunobead assay, and conjunctival goblet cells were counted by periodic acid-Schiff staining at 10 days after treatment.

Results

Both the TCP mixture groups indicated a significant improvement in TBUT, ROS production, and MDA concentrations compared to those in the DQS alone group. Furthermore, the 0.01% TCP mixture group also showed higher tear film lipid layer grades and conjunctival goblet cell density and lower corneal fluorescein staining scores, number of CD4 + IFN-γ+ T cells, and levels of TNF-α, IL-1β, and CCL4 than the DQS alone group (P < 0.05).

Conclusions

Application of eye drops containing the mixture of DQS and TCP could stabilize the tear film lipid layer, improve TBUT and corneal epithelial damages, decrease ROS production, inflammatory molecules, and T cells, and increase conjunctival goblet cell density on the ocular surface. Topical DQS and TCP mixtures may have a greater therapeutic effect on clinical signs, oxidative damage, and inflammation of dry eye than DQS eye drops.

中文翻译：

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含有3%地夸磷索钠和醋酸生育酚（维生素E）混合物的滴眼液对小鼠干眼眼表的影响

摘要

目的

研究在实验性干眼 (EDE) 小鼠模型中局部应用 3% 地夸磷索钠 (DQS) 和生育酚 (TCP) 醋酸盐混合物的疗效。

方法

暴露于干燥压力5天后，使用由3% DQS单独、0.01% TCP单独或3% DQS和0.005%或0.01% TCP混合物组成的眼药水治疗EDE。在治疗后 0、5 和 10 天测量泪液体积、泪膜破裂时间 (TBUT)、角膜荧光素染色评分 (CFSS) 和泪膜脂质层等级 (TFLLG)。用于生产活性氧 (ROS) 的 2',7'-二氯二氢荧光素二乙酸酯测定 (DCFDA)、用于丙二醛 (MDA) 的酶联免疫吸附测定 (ELISA) 和用于 CD4 + 干扰素 (IFN)-γ+ T 的流式细胞术在治疗后10天在眼表上评估细胞。此外，肿瘤坏死因子 (TNF)-α、白细胞介素 (IL)-1β、IL-6、

结果

与单独的 DQS 组相比，两个 TCP 混合物组都表明 TBUT、ROS 产生和 MDA 浓度有显着改善。此外，0.01% TCP 混合物组还表现出更高的泪膜脂质层等级和结膜杯状细胞密度以及更低的角膜荧光素染色评分、CD4 + IFN-γ + T 细胞数量和 TNF-α、IL-1β 水平，和 CCL4 比 DQS 单独组 ( P  < 0.05)。

结论

应用含有 DQS 和 TCP 混合物的滴眼液可以稳定泪膜脂质层，改善 TBUT 和角膜上皮损伤，减少 ROS 产生、炎症分子和 T 细胞，并增加眼表结膜杯状细胞密度。与 DQS 滴眼液相比，外用 DQS 和 TCP 混合物对干眼的临床症状、氧化损伤和炎症的治疗效果可能更大。