Department of Ophthalmology, the First Affiliated Hospital of Soochow University, Suzhou, , Jiangsu Province, P.R. China.
Department of Ophthalmology, the Affiliated Hospital of XuZhou Medical University, Xuzhou, Jiangsu Province, P.R. China.
Invest Ophthalmol Vis Sci. 2022 Jan 3;63(1):39. doi: 10.1167/iovs.63.1.39.
The purpose of this study was to introduce a novel dry eye rat model induced by aerosol exposure of particulate matter (PM).
A total of 30 female Sprague Dawley (SD) rats divided into 3 groups: the control group, the low-level exposed group, and the high-level exposed group. The rats in the experience groups were directly exposed to PM samples in the exposure chamber over 14 days. The clinical observation, including tear volume, corneal fluorescein staining, breakup time (BUT), inflammation index, corneal irregularity score, and corneal confocal microscopy. Eyeballs were collected on day 14 for hematoxylin and eosin (H&E) staining and PAS staining. TUNEL assay, CD45, and Ki67 immunostaining was performed and corneal ultrastructural changes were detected by electron microscopy. IL-1β, TNF-α, IFN-γ, and NF-κB Western blot analysis were used to observe the possible pathogenesis.
In the PM-treated groups, the number of layers in the corneal epithelium and corneal nerve fiber length were significantly decreased compared with that of the control group. The number of corneal epithelial microvilli and chondriosome/desmosomes were drastically reduced in PM-treated groups. Confocal microscopy and CD45 immunohistochemistry showed inflammatory cell infiltration in the PM-treated groups. PM caused apoptosis of corneal and conjunctival epithelial cells while leading to abnormal epithelial cell proliferation, meanwhile, conjunctival goblet cells in the PM-treated group were also significantly reduced. PM significantly increased the levels of IL-1β, TNF-α, IFN-γ, and p-NF-κB-p65 in the cornea.
Aerosol exposure of PM can reduce the stability of tear film and cause the change of ocular surface, which is similar to the performance of human dry eye, suggesting a novel animal model of dry eye.
本研究旨在介绍一种通过气溶胶暴露颗粒物(PM)诱导的新型干眼症大鼠模型。
将 30 只雌性 Sprague Dawley(SD)大鼠分为 3 组:对照组、低水平暴露组和高水平暴露组。实验组大鼠在暴露室内直接暴露于 PM 样品 14 天。临床观察包括泪液量、角膜荧光素染色、泪膜破裂时间(BUT)、炎症指数、角膜不规则评分和角膜共聚焦显微镜。第 14 天收集眼球进行苏木精和伊红(H&E)染色和 PAS 染色。进行 TUNEL 检测、CD45 和 Ki67 免疫染色,并通过电子显微镜检测角膜超微结构变化。采用 IL-1β、TNF-α、IFN-γ 和 NF-κB Western blot 分析观察可能的发病机制。
在 PM 处理组中,与对照组相比,角膜上皮层的层数和角膜神经纤维长度明显减少。PM 处理组角膜上皮微绒毛和桥粒/半桥粒数量急剧减少。共聚焦显微镜和 CD45 免疫组化显示 PM 处理组有炎症细胞浸润。PM 导致角膜和结膜上皮细胞凋亡,同时导致上皮细胞异常增殖,此外,PM 处理组结膜杯状细胞也明显减少。PM 显著增加了角膜中 IL-1β、TNF-α、IFN-γ 和 p-NF-κB-p65 的水平。
气溶胶暴露 PM 可降低泪膜稳定性并引起眼表面变化,类似于人类干眼症的表现,提示一种新型干眼症动物模型。