Li Lan, Jin Rujun, Li Ying, Yoon Hee Su, Yoon Hyeon Jeong, Yoon Kyung Chul
Department of Ophthalmology, Chonnam National University Medical School and Hospital, Gwangju, South Korea.
Department of Biomedical Sciences and Center for Creative Biomedical Scientists, Chonnam National University, Gwangju, South Korea.
Cutan Ocul Toxicol. 2021 Dec;40(4):350-358. doi: 10.1080/15569527.2021.1973022. Epub 2021 Sep 8.
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).
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.
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 ( < 0.05).
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.
在实验性干眼（EDE）小鼠模型中研究局部应用3%双醋瑞因钠（DQS）和醋酸生育酚（TCP）混合物的疗效。
在暴露于干燥应激5天后，应用单独含3%DQS、单独含0.01%TCP或含3%DQS与0.005%或0.01%TCP混合物的滴眼液治疗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）。在治疗后10天，对眼表进行用于检测活性氧（ROS）产生的2',7'-二氯二氢荧光素二乙酸酯检测（DCFDA）、用于检测丙二醛（MDA）的酶联免疫吸附测定（ELISA）以及用于检测CD4 +干扰素（IFN）-γ+ T细胞的流式细胞术评估。此外，在治疗后10天，使用多重免疫珠测定法测量结膜中肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β、IL-6和趋化因子CC基序配体4（CCL4）的水平，并通过过碘酸-希夫染色对结膜杯状细胞进行计数。
与单独使用DQS的组相比，两个TCP混合物组在TBUT、ROS产生和MDA浓度方面均有显著改善。此外，0.01%TCP混合物组与单独使用DQS的组相比，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及CCL4水平（P < 0.05）。
应用含DQS和TCP混合物的滴眼液可稳定泪膜脂质层，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。局部应用DQS和TCP混合物对干眼的临床体征、氧化损伤和炎症可能比DQS滴眼液具有更大的治疗效果。
