State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, China.
Eye Center, Zhongshan Hospital of Sun Yat-Sen University, Zhongshan, Guangdong, China.
Dis Markers. 2022 Jul 8;2022:1380560. doi: 10.1155/2022/1380560. eCollection 2022.
Fungal keratitis, one of the most common infectious eye diseases in China, often results in a poor prognosis due to a delayed diagnosis and the insufficiency of effective therapy. There is an urgent need to identify specific biomarkers for the disease. In this study, we screened out tear proteins in patients with fungal keratitis by microsphere-based immunoassay analysis. Levels of cytokine expression were determined in both human corneal epithelial cell models in vitro and the corneas of patients by western blot, quantitative polymerase chain reaction (qPCR), and immunofluorescence analysis. Neutrophil activation was examined by flow cytometry analysis. The relationship between the cytokine expression and neutrophils was evaluated by immunofluorescence costaining and correlation analysis. These results demonstrated that the galectin-3 expression level was increased in both cell model and patient samples at the early and late stages of fungal keratitis. The neutrophils were significantly activated during the disease course of fungal keratitis. Meanwhile, colocalization and a positive correlation between galectin-3 and neutrophils were observed, suggesting that galectin-3 may play a crucial role in the recruitment of neutrophils and immune regulation of fungal keratitis. In conclusion, galectin-3 could be a key disease marker implying a beneficial immune response in the pathogenesis of fungal keratitis, which might be a target of therapeutic strategy in the future.
真菌性角膜炎是中国最常见的感染性眼病之一,由于诊断延误和有效治疗不足,往往预后不良。因此,迫切需要确定该疾病的特定生物标志物。在本研究中,我们通过基于微球的免疫分析筛选了真菌性角膜炎患者的泪液蛋白。通过 Western blot、定量聚合酶链反应 (qPCR) 和免疫荧光分析,在体外人角膜上皮细胞模型和患者的角膜中测定细胞因子的表达水平。通过流式细胞术分析检测中性粒细胞的激活。通过免疫荧光共染色和相关分析评估细胞因子表达与中性粒细胞之间的关系。这些结果表明,在真菌性角膜炎的早期和晚期,细胞模型和患者样本中 galectin-3 的表达水平均升高。中性粒细胞在真菌性角膜炎的病程中明显被激活。同时,观察到 galectin-3 与中性粒细胞之间的共定位和正相关,表明 galectin-3 可能在中性粒细胞的募集和真菌性角膜炎的免疫调节中发挥关键作用。总之,galectin-3 可能是真菌性角膜炎发病机制中的关键疾病标志物,暗示有益的免疫反应,可能成为未来治疗策略的靶点。