Zhang Zhen, Yang Wen-Zhao, Zhu Zhen-Zhen, Hu Qian-Qian, Chen Yan-Feng, He Hui, Chen Yong-Xiong, Liu Zu-Guo
Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China.
Invest Ophthalmol Vis Sci. 2014 May 6;55(5):2963-74. doi: 10.1167/iovs.13-13577.
We investigated the therapeutic effects and underlying mechanisms of topical doxycycline in a benzalkonium chloride (BAC)-induced mouse dry eye model.
Eye drops containing 0.025%, 0.1% doxycycline or solvent were administered to a BAC-induced dry eye model four times daily. The clinical evaluations, including tear break-up time (BUT), fluorescein staining, inflammatory index, and tear volume, were performed on days 0, 1, 4, 7, and 10. Global specimens were collected on day 10 and processed for immunofluorescent staining, TUNEL, and periodic acid-Schiff assay. The levels of inflammatory mediators in the corneas were determined by real-time PCR. The total and phosphorylated nuclear factor-κB (NF-κB) were detected by Western blot.
Both 0.025% and 0.1% doxycycline treatments resulted in increased BUT, lower fluorescein staining scores, and inflammatory index on days 4, 7, and 10, while no significant change in tear volume was observed. The 0.1% doxycycline-treated group showed more improvements in decreasing fluorescein staining scores, increasing Ki-67-positive cells, and decreasing TUNEL- and keratin-10-positive cells than other groups. The mucin-filled goblet cells in conjunctivas were increased, and the expression of CD11b and levels of matrix metalloproteinase-9, IL-1β, IL-6, TNF-α, macrophage inflammatory protein-2, and cytokine-induced neutrophil chemoattractant in corneas were decreased in both doxycycline-treated groups. In addition, doxycycline significantly reduced the phosphorylation of NF-κB activated in the BAC-treated corneas.
Topical doxycycline showed clinical improvements and alleviated ocular surface inflammation on BAC-induced mouse dry eye, suggesting a potential as an anti-inflammatory agent in the clinical treatment of dry eye.
我们研究了局部应用强力霉素在苯扎氯铵（BAC）诱导的小鼠干眼模型中的治疗效果及潜在机制。
将含有0.025%、0.1%强力霉素或溶剂的滴眼液每日4次给予BAC诱导的干眼模型。在第0、1、4、7和10天进行临床评估，包括泪膜破裂时间（BUT）、荧光素染色、炎症指数和泪液量。在第10天收集整体标本并进行免疫荧光染色、TUNEL和过碘酸希夫氏反应检测。通过实时PCR测定角膜中炎症介质的水平。通过蛋白质印迹法检测总核因子-κB（NF-κB）和磷酸化核因子-κB。
0.025%和0.1%强力霉素治疗均使第4、7和10天的BUT增加，荧光素染色评分和炎症指数降低，而泪液量未观察到显著变化。与其他组相比，0.1%强力霉素治疗组在降低荧光素染色评分、增加Ki-67阳性细胞以及减少TUNEL和角蛋白-10阳性细胞方面有更多改善。两个强力霉素治疗组结膜中充满黏蛋白的杯状细胞均增加，角膜中CD11b的表达以及基质金属蛋白酶-9、IL-1β、IL-6、TNF-α、巨噬细胞炎性蛋白-2和细胞因子诱导的中性粒细胞趋化因子的水平均降低。此外，强力霉素显著降低了BAC处理的角膜中活化的NF-κB的磷酸化水平。
局部应用强力霉素在BAC诱导的小鼠干眼中显示出临床改善并减轻了眼表炎症，提示其在干眼临床治疗中作为抗炎药物的潜力。
